Publications by authors named "Lingling Zu"

24 Publications

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Prognostic value of ferroptosis-related genes in patients with lung adenocarcinoma.

Thorac Cancer 2021 Jun 12;12(12):1890-1899. Epub 2021 May 12.

Department of Lung Cancer Surgery, Tianjin Medical University General Hospital, Tianjin, China.

Background: The prevalence of lung adenocarcinomas (LUADs) has dramatically increased in recent decades. Ferroptosis is a process of iron-dependent regulatory cell death. It is still unclear whether the expression of ferroptosis-related genes (FRGs) is involved in the pathogenesis and survival of patients with LUAD.

Methods: We retrieved LUAD data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases and used LASSO Cox regression analysis to select the gene signature suitable for modeling. The risk score was calculated according to the model, and the patients were divided into high- and low-risk groups according to the median risk score. Functional enrichment analysis was carried out by this group, and a model for predicting clinical prognosis was established by combining this group with clinical factors.

Results: Gene set enrichment analysis (GSEA) and single-sample gene set enrichment analysis (ssGSEA) analysis showed that there were several immune-related pathways and immune infiltration differences between high- and low-risk groups. A prognostic model integrating 10 ferroptosis-related genes (FR-DEGs), and clinical factors were constructed and validated in an external cohort.

Conclusions: The FR-DEGs signature was related to immune infiltration, and a model based on FR-DEGs and clinical factors was established to predict the prognosis of patients with LUAD.
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http://dx.doi.org/10.1111/1759-7714.13998DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8201541PMC
June 2021

Mutations Associated with No Durable Clinical Benefit to Immune Checkpoint Blockade in Non-S-Cell Lung Cancer.

Cancers (Basel) 2021 Mar 19;13(6). Epub 2021 Mar 19.

Department of Lung Cancer Surgery, Tianjin Medical University General Hospital, Tianjin 300050, China.

(1) Background: The immune checkpoint blockade (ICB) has shown promising efficacy in non-small-cell lung cancer (NSCLC) patients with significant clinical benefits and durable responses, but the overall response rate to ICBs is only 20%. The lack of responsiveness to ICBs is currently a central problem in cancer immunotherapy. (2) Methods: Four public cohorts comprising 2986 patients with NSCLC were included in the study. We screened 158 patients with NSCLC with no durable clinical benefit (NDB) to ICBs in the Rizvi cohort and identified NDB-related gene mutations in these patients using univariate and multivariate Cox regression analyses. Programmed death-ligand 1 (PD-L1) expression, tumor mutation burden (TMB), neoantigen load, tumor-infiltrating lymphocytes, and immune-related gene expression were analyzed for identifying gene mutations. A comprehensive predictive classifier model was also built to evaluate the efficacy of ICB therapy. (3) Results: Mutations in FAT1 and KEAP1 were found to correlate with NDB in patients with NSCLC to ICBs; however, the analysis suggested that only mutation in FAT1 was valuable in predicting the efficacy of ICB therapy, and that mutation in KEAP1 acted as a prognostic but not a predictive biomarker for NSCLC. Mutations in FAT1 were associated with a higher TMB and lower multiple lymphocyte infiltration, including CD8 (T-Cell Surface Glycoprotein CD8)+ T cells. We established a prognostic model according to PD-L1 expression, TMB, smoking status, treatment regimen, treatment type, and FAT1 mutation, which indicated good accuracy by receiver operating characteristic (ROC) analysis (area under the curve (AUC) for 6-months survival: 0.763; AUC for 12-months survival: 0.871). (4) Conclusions: Mutation in FAT1 may be a predictive biomarker in patients with NSCLC who exhibit NDB to ICBs. We proposed an FAT1 mutation-based model for screening more suitable NSCLC patients to receive ICBs that may contribute to individualized immunotherapy.
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http://dx.doi.org/10.3390/cancers13061397DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8003499PMC
March 2021

Deubiquitinating enzyme USP41 promotes lung cancer cell proliferation and migration.

Thorac Cancer 2021 04 22;12(7):1041-1047. Epub 2021 Feb 22.

Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin, China.

Background: To reveal the function of deubiquitylating enzyme USP41 in lung adenocarcinoma.

Methods: The relationship between USP41 and lung cancer was determined by analyzing data from The Cancer Genome Atlas (TCGA). A549 and H1299 cell lines were transfected with short hairpin RNA against USP41 (shUSP41 group) or negative control (shCon group). Western blotting was used to verify the transfection efficacy and marker expression. Cell proliferation and apoptosis were analyzed by EdU assay, MTT assay, and flow cytometry after USP41 knockdown. Transwell assay was used to determine the effect of USP41 downregulation on cell migration.

Results: Analysis of lung cancer data from TCGA database indicated a higher level of USP41 expression in lung cancer tumor tissue compared with that in noncancerous tissue, and USP41 overexpression was correlated with poor overall survival of lung cancer patients (p < 0.01). The outcomes of the EdU, MTT, and flow cytometry assays indicated decreased cell proliferation and enhanced apoptosis in shUSP41-transfected cells. Transwell assay further demonstrated that USP41 knockdown increased the migration rate of A549 and H1299 cells.

Conclusions: In our study, USP41 was overexpressed in lung cancer tissue and associated with poor prognosis of lung cancer. USP41 knockdown inhibits cell proliferation and migration and induces cell apoptosis of lung cancer.
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http://dx.doi.org/10.1111/1759-7714.13843DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8017261PMC
April 2021

The first complete mitochondrial genome of Blossom-headed Parakeet (Psittaciformes: Psittacidae).

Mitochondrial DNA B Resour 2020 Sep 29;5(3):3457-3459. Epub 2020 Sep 29.

Key Laboratory for Conserving Wildlife with Small Populations in Yunnan, Southwest Forestry University, Kunming, China.

Blossom-headed Parakeet is listed as Near Threatened on the IUCN Red List of Threatened Species because of its habitat loss, and declining population. In this study, we first sequenced and described the complete mitochondrial genome and phylogeny of . The whole genome of was 16,814 bp in length, and contained 14 protein-coding genes, 22 transfer RNA genes, 2 ribosome RNA genes, and 1 non-coding control regions. The overall base composition of the mitochondrial DNA was 31.88% for A, 22.00% for T, 32.88% for C, 13.23% for G, with a GC content of 46.11%. A phylogenetic tree strongly supported that genus closely related with genus by highly probability.
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http://dx.doi.org/10.1080/23802359.2020.1825131DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7782874PMC
September 2020

Nm23-H1 inhibits lung cancer bone-specific metastasis by upregulating miR-660-5p targeted SMARCA5.

Thorac Cancer 2020 03 5;11(3):640-650. Epub 2020 Feb 5.

Lung Cancer Center, West China Hospital, Sichuan University, Chengdu, China.

Background: Nm23-H1 gene has been found to be an inhibitor of tumor metastasis in lung cancer. MicroRNAs (miRNAs) play key roles in tumor metastasis through multiple signaling pathways. This study explored whether the nm23-H1 gene could inhibit invasion and metastasis of lung cancer cells by regulating miRNA-660-5p targets.

Methods: Quantitative real-time PCR (qRT-PCR) and western blots were used to measure the expression of nm23-H1 and miR-660-5p of various human lung cancer cell lines. Cell counting kit-8 (CCK-8), wound-healing and transwell assay were carried out to assess cell proliferation, migration and invasion of each cell line. Xenograft were applied to determine in vivo effects of miR-660-5p among nude mice. Luciferase assay and western blot were performed to determine the target gene of miR-660-5p.

Results: We found that high expression of nm23-H1 correlated with decreased miRNA-660-5p expression. Inhibiting miR-660-5p suppressed lung cancer cells progression significantly in vitro, whereas overexpression of miR-660-5p facilitated tumor growth and bone metastasis in vivo. In addition, as the potential target gene of miR-660-5p, SMARCA5 overexpression in vitro suppressed tumor progression and osteolytic metastasis associated RANKL signaling, which is congruent with the effect of nm23-H1 on the lung cancer cells.

Conclusion: Nm23-H1 inhibits tumor progression and bone-specific metastasis of lung cancer by regulating miR-660-5p/SMARCA5/RANKL axis, which indicates the related genes may serve as potential targets for the treatment of human lung cancer.

Key Points: Significant findings of the study High expression of nm23-H1 correlated with decreased miRNA-660-5p expression. Further, downregulation of miR-660-5p significantly suppressed the tumor progression and bone-specific metastasis of lung cancer cells. What this study adds This is the first study to show an inverse association between nm23-H1 and miR-660-5p, and confirm that nm23-H1 inhibits tumor progression and bone-specific metastasis of lung cancer by regulating miR-660-5p/SMARCA5/RANKL axis.
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http://dx.doi.org/10.1111/1759-7714.13308DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7049508PMC
March 2020

Comparative mitochondrial proteomic analysis of human large cell lung cancer cell lines with different metastasis potential.

Thorac Cancer 2019 05 4;10(5):1111-1128. Epub 2019 Apr 4.

Lung Cancer Center/Lung Cancer Institute, West China Hospital, Sichuan University, Chengdu, China.

Background: Lung cancer is a highly aggressive cancer with a poor prognosis and is associated with distant metastasis; however, there are no clinically recognized biomarkers for the early diagnosis and prediction of lung cancer metastasis. We sought to identify the differential mitochondrial protein profiles and understand the molecular mechanisms governing lung cancer metastasis.

Methods: Mitochondrial proteomic analysis was performed to screen and identify the differential mitochondrial protein profiles between human large cell lung cancer cell lines with high (L-9981) and low (NL-9980) metastatic potential by two-dimensional differential gel electrophoresis. Western blot was used to validate the differential mitochondrial proteins from the two cells. Bioinformatic proteome analysis was performed using the Mascot search engine and messenger RNA expression of the 37 genes of the differential mitochondrial proteins were detected by real-time PCR.

Results: Two hundred and seventeen mitochondrial proteins were differentially expressed between L-9981 and NL-9980 cells (P < 0.05). Sixty-four analyzed proteins were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry coupled with database interrogation. Ontology analysis revealed that these proteins were mainly involved in the regulation of translation, amino acid metabolism, tricarboxylic acid cycle, cancer invasion and metastasis, oxidative phosphorylation, intracellular signaling pathway, cell cycle, and apoptosis.

Conclusion: Our results suggest that the incorporation of more samples and new datasets will permit the definition of a collection of proteins as potential biomarkers for the prediction and diagnosis of lung cancer metastasis.
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http://dx.doi.org/10.1111/1759-7714.13052DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6501018PMC
May 2019

The RNA-binding protein Sam68 is critical for non-small cell lung cancer cell proliferation by regulating Wnt/β-catenin pathway.

Int J Clin Exp Pathol 2017 1;10(8):8281-8291. Epub 2017 Aug 1.

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital Tianjin, P. R. China.

Src associated in mitosis, 68 kDa (Sam68) is a KH domain RNA-binding protein that regulates a broad scope of biological events, including RNA metabolism, transcription and signal transduction. Herein, we aimed to explore the expression, clinical significance and biological function of Sam68 in human non-small cell lung cancer (NSCLC). By applying quantitative real-time PCR (qRT-PCR), western blotting and immunohistochemistry (IHC) methods, we found that nucleic localized Sam68 was markedly overexpressed in NSCLC tissues and cell lines. By X analysis and Kaplan-Meier survivial analysis between Sam68 expression and various clinicopathological features, Sam68 was found to be significantly associated with clinical T stage, advanced tumor grade, and short overall survival. Finally, loss-of-function studies showed that knockdown of Sam68 inhibited cell proliferation, colony formation and cell cycle progression in NSCLC cells. Moreover, our results clarified that knockdown of Sam68 could suppress NSCLC cell proliferation via the inhibition of Wnt/β-catenin pathway. To conclude, our results demonstrated that upregulation of Sam68 in NSCLC resulted in poor prognosis, and it promoted cell proliferation via activating Wnt/β-catenin signaling pathway, which could serve as a novel biomarker for the prognosis and therapy of NSCLC.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6965395PMC
August 2017

Activating transcription factor 3 promotes malignance of lung cancer cells in vitro.

Thorac Cancer 2017 05 27;8(3):181-191. Epub 2017 Feb 27.

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin, China.

Background: Lung cancer remains the most common cause of cancer-related death, with high rates of recurrence and poor outcomes. An abnormally high expression of activating transcription factor 3 (ATF3) in various cancers suggests an oncogenic role; however, its function in lung cancer is largely unknown.

Methods: Sixty-four pairs of lung cancer tissues were collected for ATF3 expression analysis by quantitative real-time PCR, immunoblotting, and immunohistochemistry staining. Correlations between ATF3 expression with clinicopathological features and overall survival were analyzed. ATF3 expression in a panel of lung cancer cell lines together with normal bronchial epithelial Beas-2B cells was also determined. Human H1299 and A549 cells were used for ATF3 knockdown and/or overexpression assays. Alterations in cell proliferation, cell cycle attribution, migration, and invasion were all assessed in vitro.

Results: Increased ATF3 messenger RNA and protein expression were observed in lung cancer tissues/cells compared with normal tissues/cells. High tumorous ATF3 expression was significantly correlated with positive advanced tumor grade, lymph node metastasis, and shorter overall survival. Experimentally, we found that RNA interference mediated knockdown of ATF3 significantly inhibited the cell proliferation, cell cycle progression, migration, and invasion capacities of lung cancer cells in vitro, whereas forced expression of ATF3 did the opposite.

Conclusion: Upregulation of ATF3 in lung cancer promotes cell proliferation, migration, and invasion, and may represent a novel therapeutic target for lung cancer.
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http://dx.doi.org/10.1111/1759-7714.12421DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5415490PMC
May 2017

Metformin potentiates anti-tumor effect of resveratrol on pancreatic cancer by down-regulation of VEGF-B signaling pathway.

Oncotarget 2016 Dec;7(51):84190-84200

Department of Histology and Embryology, School of Medicine, Nankai University, Tianjin, China.

Our previous study showed that resveratrol (RSV) exhibited not only anti-tumor effect, but also had potential tumor promotion effect on pancreatic cancer (Paca) cells through up-regulation of VEGF-B. We determined whether metformin (MET) could potentiate the anti-tumor effect of RSV on PaCa in this study. Combination of RSV (100 μmol/l) and MET (20 mmol/l) significantly inhibited tumor growth and increased apoptosis of human PaCa in comparison with RSV or MET alone treatment in PaCa cell lines (Miapaca-2, Panc-1 and Capan-2). Combination of RSV (60 mg/kg, gavage) and MET (250 mg/kg, i.p.) significantly inhibited tumor growth in PaCa bearing nude mice (subcutaneous injection of 5 × 106 Miapaca-2 cells) in comparison with RSV or MET alone treatment on day 40. Combination treatment significantly decreased VEGF-B expression and inhibited activity of GSK-3β when compared to the RSV alone treatment. Up-regulated expressions of Bax, cleaved caspase-3 and down-regulated expression of Bcl-2 were observed in RSV+ MET group in comparison with RSV group either in vitro or in vivo. Inhibition of VEGF-B by VEGF-B small interfering RNA (siRNA) mimicked the effects of MET on PaCa cells. These results suggested that MET, a potential pharmacological inhibitor of VEGF-B signaling pathway, potentiated the anti-tumor effect of RSV on PaCa, and combination of MET and RSV would be a promising modality for clinical PaCa therapy.
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http://dx.doi.org/10.18632/oncotarget.12391DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5356654PMC
December 2016

WW45, a Gli1 binding protein, negatively regulated Hedgehog signaling in lung cancer.

Oncotarget 2016 10;7(42):68966-68975

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Environment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin 300052, China.

Over-expression of Gli1 is very common in lung cancer. However, the underlying molecular mechanism remains largely unknown. Here, using mass spectrum, we have identified WW45 as a binding partner of Gli1. WW45 interacted with Gli1, promoted its ubiquitination and inhibited the expression of its target genes. In the functional studies, WW45 inhibited the growth and migration of lung cancer cells. Knocking down the expression of WW45 promoted the growth and migration of lung cancer cells, which was rescued by down-regulation of Gli1. Moreover, over-expression of WW45 inhibited the tumorigenesis in a de novo lung cancer tumorigenesis mouse model (LKB-Ras) as well as the expression of Gli1. Also over-expression of WW45 improved the survival of these mice. In addition, the expression of WW45 was down-regulated in the clinical lung cancer samples, which was inversely correlated with the expression of Gli1. Taken together, this study demonstrated the suppressive roles of WW45 in lung cancer by inhibiting the Hedgehog/Gli1 signaling.
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http://dx.doi.org/10.18632/oncotarget.12155DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5356604PMC
October 2016

Erratum: MicroRNA-338-3p suppresses metastasis of lung cancer cells by targeting the EMT regulator Sox4.

Am J Cancer Res 2016 1;6(7):1582. Epub 2016 Jul 1.

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital Tianjin 300052, China.

[This corrects the article on p. 127 in vol. 6, PMID: 27186391.].
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4969407PMC
August 2016

Nm23-H1 was involved in regulation of KAI1 expression in high-metastatic lung cancer cells L9981.

J Thorac Dis 2016 Jun;8(6):1217-26

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin 300052, China.

Background: The tetraspanin KAI1/CD82 was identified as a tumor metastasis suppressor that down-regulated in malignant progression of lung cancer. However, the underlying mechanism of anti-metastasis role of KAI1 in lung cancer is hardly known. In this paper, we sought to study the function and regulatory mechanism of KAI1 in high metastasis lung cancer cell line.

Methods: KAI1 expression was detected in high/low metastatic large lung cancer cell line L9981/NL9980 by quantitative real-time polymerase chain reaction (qRT-PCR). The tumor suppressor function of KAI1 was determined by wound healing assay after over-expression or knockdown of KAI1 in L9981 or NL9980 cells. Invasion assay was performed to detect the invasion ability of L9981 by transfection of KAI1. The effect of tumor suppressor p53 on KAI1 expression was measured by western blot and luciferase assay. Then the regulation of KAI1 due to over-expression of metastasis suppressor nm23-H1 was monitored by qRT-PCR, western blot and reporter gene assay. The progression of L9981 cells after p53 and nm23-H1 expression was detected by invasion assay. Also, methylation status of KAI1 promoter in NL9980 and L9981 cells were examined by bisulfite sequencing and methylation-specific PCR.

Results: We found that KAI1 is down-regulated in high metastatic L9981 cells compare with NL9980 cells. The migration and invasion of L9981 cells were remarkably suppressed in vitro by KAI1 transfection. The migration ability of NL9980 was enhanced by inhibition of KAI1. Furthermore, KAI1 expression was induced after over-expression of p53 or nm23-H1, while cell invasion was inhibited in L9981 cells. The results of reporter analysis indicated that KAI1 promoter region between -922 to -846 could response to nm23-H1. In addition, we discovered only slight methylation of KAI1 promoter, which showed that loss expression of KAI1 in L9981 cells may not due to promoter methylation.

Conclusions: The results suggested that nm23-H1 was involved in the KAI1-regulated inhibition of metastasis in lung cancer cells. More insights into the relationship between KAI1 and other metastasis suppressors will pave the way for the elucidation of anti-metastasis mechanism in lung cancer.
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http://dx.doi.org/10.21037/jtd.2016.04.59DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4885971PMC
June 2016

MicroRNA-338-3p suppresses metastasis of lung cancer cells by targeting the EMT regulator Sox4.

Am J Cancer Res 2016 15;6(2):127-40. Epub 2016 Jan 15.

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital Tianjin 300052, China.

Metastasis remains the leading cause of the majority of cancer-related mortality. MicroRNAs (miRNAs) have frequently emerged as tumor metastatic regulator by acting on multiple signaling pathways. In the present study, we demonstrated that miR-338-3p was significantly downregulated in highly metastatic NSCLC cell lines and clinical metastatic tissues. Then, we found that introduction of miR-338-3p significantly suppressed the migration and invasion of lung cancer cells both in vitro and in vivo, suggesting that miR-338-3p may be a novel tumor suppressor. Further studies indicated that the EMT-related transcription factor Sox4 was one direct target gene of miR-338-3p, evidenced by the direct binding of miR-338-3p with the 3'untranslated region (3'UTR) of Sox4. Furthermore, miR-338-3p could decrease the expression of Sox4 both at mRNA and protein levels. Notably, the EMT marker E-cadherin or vimentin, a downstream regulator of Sox4, was also down-regulated or up-regulated upon miR-338-3p treatment. Additionally, over-expressing or silencing Sox4 could elevate or inhibit the migration and invasion of lung cancer cells, parallel to the effect of miR-338-3p on the lung cancer cells. Meanwhile, knockdown of Sox4 reversed the enhanced migration and invasion mediated by miR-338-3p. These results indicated that miR-338-3p suppressed the migration and invasion of NSCLC cells through targeting Sox4 involving in the EMT process. Thus, our finding provides new insight into the mechanism of NSCLC progression. Therapeutically, miR-338-3p may serve as a potential target in the treatment of human lung cancer.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4859648PMC
May 2016

[Nicotine Induced Lung Cancer Cells Epithelial-mesenchymal Transition 
and Promote Its Vitro Invasion Potential].

Zhongguo Fei Ai Za Zhi 2016 Apr;19(4):169-76

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin 300052, China;Lung Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.

Background: Our previous study found that nicotine could induce lung cancer cell epithelial-mesenchymal transition (EMT). The aim of this study is to explore the relationship between nicotine-induced EMT and lung cancer invasion and metastasis.

Methods: Real-time PCR and Western blot were used to detect the expression changes of EMT-related markers, E-cadherin and Vimentin, in A549 lung cancer cells treated with nicotine; The transposition of β-catenin protein expression was determined by immunofluorescence; Scratch test and Transwell invasion assay were used to detect the effects of nicotine on lung cancer cell migration and invasion.

Results: Nicotine can significantly down-regulate the expressional level of E-cadherin mRNA and protein of A549 cells in a manner of dose and time-dependent (P<0.01, P<0.01); Nicotine can significantly up-regulate the expressional level of Vimentin mRNA and protein of A549 cells in a manner of dose and time-dependent (P<0.01, P<0.01); Immunofluorescence results showed that β-catenin protein was significantly transfered to nucleus; Scratch test and Transwell assay showed that Nicotine could remarkably increase the migration and invasion potential of lung cancer cells (P<0.01, P<0.01).

Conclusions: Nicotine can induce cancer cells EMT, and promote the invasion and metastasis ability of lung cancer cells.
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http://dx.doi.org/10.3779/j.issn.1009-3419.2016.04.11DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5999818PMC
April 2016

Anti-apoptotic brain and reproductive organ-expressed proteins enhance cisplatin resistance in lung cancer cells via the protein kinase B signaling pathway.

Thorac Cancer 2016 Mar 20;7(2):190-8. Epub 2015 Oct 20.

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute Tianjin Medical University General Hospital Tianjin China.

Background: Cisplatin-based chemotherapy is the standard first-line treatment for non-small-cell lung cancers (NSCLCs); however, the long-term therapeutic effect is reduced by chemoresistance. Brain and reproductive organ-expressed (BRE) proteins are overexpressed in several cancers and have an anti-apoptotic function. However, their biological role in the development of the chemoresistant phenotype of human NSCLC remains unknown. We investigate the differential expression of the BRE gene in human lung adenocarcinoma cell lines A549 and the cisplatin-resistant variant A549/cisplatin (DDP), and the mechanisms of cisplatin-resistance induced by the BRE gene.

Methods: Cell counting kit-8 assay was employed to determine the sensitivity of A549 and A549/DDP cell lines to cisplatin. BRE expression was measured using quantitative real time-polymerase chain reaction and western blot analysis. The apoptosis rate of lung adenocarcinoma cells was determined by flow cytometry.

Results: BRE expression in A549 cells, derived from human lung cells, was markedly decreased compared with parental cisplatin-resistant A549/DDP cells at messenger ribonucleic acid and protein levels. BRE overexpression in A549 significantly decreased sensitivity to DDP by inhibiting cell apoptosis. Conversely, BRE knockdown in A549/DDP cells increased their chemosensitivity. Importantly, we demonstrate that BRE overexpression induces the expression of phosphoprotein kinase B (p-Akt) in lung cancer cells, while BRE silencing inhibits p-Akt expression. Furthermore, downregulation of p-Akt by LY294002 reversed the DDP resistance induced by BRE by increasing apoptosis. BRE enhances the DDP resistance of lung cancer cells through the Akt signaling pathway.

Conclusion: Our findings provide new insight into the mechanism of DDP resistance in NSCLC cells and suggest BRE as an attractive new target for NSCLC treatment.
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http://dx.doi.org/10.1111/1759-7714.12313DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4773300PMC
March 2016

MiR-449a suppresses cell invasion by inhibiting MAP2K1 in non-small cell lung cancer.

Am J Cancer Res 2015 15;5(9):2730-44. Epub 2015 Aug 15.

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital Tianjin 300052, China.

Increasing evidence reveals that deregulation of miRNAs contributes to carcinogenesis of the human non-small cell lung cancer (NSCLC). Our study discovered that the expression of miR-449a was markedly decreased in NSCLC cells with high metastatic capacity and tissues of positive lymph node metastasis. Moreover, our results showed that miR-449a could act as a tumor suppressor by inhibiting the invasion of NSCLC cells in vitro and in vivo. Mechanistically, miR-449a inhibited the expression of MAP2K1 by direct targeting its 3'UTR, and regulated the activity of MEK1/ERK1/2/c-Jun pathway through an auto-regulatory feedback loop. Furthermore, the histone methylation mediated the decreased expression of miR-449a through SUZ12. Taken together, the novel connection between miR-449a and MAP2K1 demonstrated here provided a new, potential therapeutic target for the treatment of non-small cell lung cancer.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4633902PMC
November 2015

miR-132 inhibits lung cancer cell migration and invasion by targeting SOX4.

J Thorac Dis 2015 Sep;7(9):1563-9

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin 300052, China.

Background: Multiple MicroRNAs (miRNAs) have been identified in the development and progression of lung cancer. However, the expression and roles of miR-132 in non-small cell lung cancer (NSCLC) remain largely undefined. The aim of this study is to investigate the biological functions and its molecular mechanisms of miR-132 in human lung cancer cells.

Methods: miR-132 expression was measured in human lung cancer cell lines by quantitative real-time PCR (qRT-PCR). The cells migration and invasion ability were measured by wound healing assay and transwell assay. The influence of miR-132 on tumor progression in vivo was monitored using NSCLC xenografts in nude mice. The target gene of miR-132 was determined by luciferase assay and western blot.

Results: The expression level of miR-132 was dramatically decreased in examined lung cancer cell lines. Then, we found that introduction of miR-132 significantly suppressed the migration and invasion of lung cancer cells in vitro. Besides, miR-132 overexpression could also inhibit tumor growth in the nude mice. Further studies indicated that the sex determining region Y-box 4 (SOX4) is a target gene of miR-132. SOX4 re-introduction could reverse the anti-invasion role of miR-132.

Conclusions: Our finding provides new insight into the mechanism of NSCLC progression. Therapeutically, miR-132 may serve as a potential target in the treatment of human lung cancer.
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http://dx.doi.org/10.3978/j.issn.2072-1439.2015.09.06DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4598519PMC
September 2015

[MiR-192 confers cisplatin resistance by targeting Bim in lung cancer].

Zhongguo Fei Ai Za Zhi 2014 May;17(5):384-90

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin 300052, China.

Background And Objective: Cisplatin is the first-line drug for the chemotherapy of non-small cell lung cancer (NSCLC), but the acquired chemoresistance restricted the effect of its treatment. The aim of this study is to validate the miRNAs related to the Cisplatin resistance in lung cancer and elucidate the molecular mechanisms.

Methods: We performed miRNA microarray and RT-PCR to obtain the aberrant differential expressed miRNAs between A549 and its paired Cisplatin-resistant cell line A549/DDP cells, and then we investigated the biological functions of miR-192, which is the aberrant differential expressed miRNA. After transfection of the miR-192 into A549 cells, we measured the half inhibition concentration (IC50), cell apoptosis of the trasfectant cells, and then we used biological softwares and dual-luciferase report assay to explore the target gene of the miR-192, which was further validated by RT-PCR and Western blot.

Results: MiR-192 was highly over-expressed in A549/DDP cells , whose quantity was 37.59±0.35 fold higher than that in A549 cells. Overexpression of miR-192 in A549 cells significantly conferred resistance to Cisplatin and inhibited apoptosis. By contrast, down-expression of miR-192 in A549/DDP cells remarkably restrained the Cisplatin resistance and induced apoptosis. MiR-192 binded to Bim 3'-UTR and negatively regulated Bim expression at the post-transcriptional level in lung adenocarcinoma cells.

Conclusions: Our data suggested that miR-192 induced Cisplatin-resistance and inhibited cell apoptosis in lung cancer via negative targeting Bim expression.
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http://dx.doi.org/10.3779/j.issn.1009-3419.2014.05.04DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6000442PMC
May 2014

MiR-517a-3p accelerates lung cancer cell proliferation and invasion through inhibiting FOXJ3 expression.

Life Sci 2014 Jul 17;108(1):48-53. Epub 2014 May 17.

Endoscopy Center China-Japan Union Hospital of Jilin University, 126 Xiantai Street, Changchun 130033, China. Electronic address:

Aims: Aberrant expression of microRNAs (miRNAs) results in alterations of various biological processes (e.g., cell cycle, cell differentiation, and apoptosis) and cell transformation. Altered miRNAs expression was associated with lung carcinogenesis and tumor progression. This study aimed to investigate the function and underlying molecular events of miR-517a-3p on regulation of lung cancer cell proliferation and invasion.

Main Methods: Transfected miR-517a-3p mimics or inhibitors into 95D and 95C cells respectively, the effects of miR-517a-3p on lung cancer cell proliferation, migration, and invasion were detected. Bioinformatics software forecasted potential target genes of miR-517a-3p and dual luciferase reporter gene system and western blot verified whether miR-517a-3p regulates FOXJ3 expression directly.

Key Findings: MiR-517a-3p was differentially expressed in lung cancer 95D and 95C cell lines that have different metastatic potential. Manipulation of miR-517a-3p expression changed lung cancer cell proliferation, migration and invasion capacity. MiR-517a-3p directly regulated FOXJ3 expression by binding to FOXJ3 promoter.

Significance: This study demonstrated that miR-517a-3p promoted lung cancer cell proliferation and invasion by targeting of FOXJ3 expression.
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http://dx.doi.org/10.1016/j.lfs.2014.05.006DOI Listing
July 2014

[Screening and establishment of human lung cancer cell lines with organ-specific metastasis potential].

Zhongguo Fei Ai Za Zhi 2014 Mar;17(3):175-82

Key Laboratory of Lung Cancer Molecular Biology, West China Hospital, Sichuan University, Chengdu 610041, China.

Background: Cancer metastasis is not only the malignant marker and characteristics, but also the main cause of failure to cure and lose their life in the patients with lung cancer. Lung cancer metastasis has organ-specific characteristics. The most common sites of lung cancer metastasis are mediastinal lymph node, brain, bone, liver and adrenal gland. The aim of this study is to screen and establish lung cancer cell model with organ-specific metastasis potential with human high-metastatic large cell lung cancer cell line L9981 established by our laboratory previously, and to provide cell models for studying the mechanisms and signal regulation of organ-specific metastasis of lung cancer.

Methods: The parent lung cancer cell line, L9981-Luc, was inoculated in the armpit of nude mice. The live animal imaging system, IVIS-200, was used to detect the lung cancer organ-specific metastasis every week. When the organ-specific metastasis were established, the nude mices bearing the lung cancer were sacrificed when they became moribund. Under sterile conditions, the organs (mediastinal lymph nodes, lung, spinal column and brain) with lung cancer organ-specific metastasis were removed and the metastasized nodules were dissected free of connective tissue and blood clots, and rinsed twice with medium. The metastasized nodules were finely minced using sterile scalpel blades in medium, and the cells were seeded in tissue culture dishes. Then, the cells with organ-specific metastasis potential were reinoculated into the armpit of nude mice, respectively. This processes were repeated to establish the organ-specific metastatic sublines of L9981-Luc cell line more than 10 times. Finally, the organ-specific metastasis sublines of L9981-Luc were screened and established, which the four cell lines have the characteristics only metastasized to brian, lung, bone and mediastinal lymph node.

Results: A group of organ-specific metastasis cell lines which only metastasized to brian, lung, bone and mediastinal lymph node were successfully established through repeating reinoculatation, live animal imaging in nude mice, and screening and identification in vitro. We named the four cell lines as L9981-BoM, L9981-LuM, L9981-BrM and L9981-LnM, respectively. The L9981-BoM cell was only metastasized to bone. The l9981-LuM cell was only metastasized to lung. The L9981-BrM only metastasized to brain. The L9981-LnM cell was only metastasized to midiastinal lymph nodes.

Conclusions: A human large cell lung cancer cell model with bone, lung, brain and lymph node-specific metastasis potential was successfully established. It will be helpful to further study the molecular mechanisms and signal regulation of lung cancer organ- specific metastasis. It will be to also provide reliable cell model for developing new techniques and molecular targeting drugs of inhibiting or reversing lung cancer metastasis.
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http://dx.doi.org/10.3779/j.issn.1009-3419.2014.03.20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6019378PMC
March 2014

MiR-132 suppresses the migration and invasion of lung cancer cells via targeting the EMT regulator ZEB2.

PLoS One 2014 13;9(3):e91827. Epub 2014 Mar 13.

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin, China.

MicroRNAs (miRNAs) are small, non-coding RNAs which can function as oncogenes or tumor suppressor genes in human cancers. Emerging evidence reveals that deregulation of miRNAs contributes to the human non-small cell lung cancer (NSCLC). In the present study, we demonstrated that the expression levels of miR-132 were dramatically decreased in examined NSCLC cell lines and clinical NSCLC tissue samples. Then, we found that introduction of miR-132 significantly suppressed the migration and invasion of lung cancer cells in vitro, suggesting that miR-132 may be a novel tumor suppressor. Further studies indicated that the EMT-related transcription factor ZEB2 was one direct target genes of miR-132, evidenced by the direct binding of miR-132 with the 3' untranslated region (3' UTR) of ZEB2. Further, miR-132 could decrease the expression of ZEB2 at the levels of mRNA and protein. Notably, the EMT marker E-cadherin or vimentin, a downstream of ZEB2, was also down-regulated or up-regulated upon miR-132 treatment. Additionally, over-expressing or silencing ZEB2 was able to elevate or inhibit the migration and invasion of lung cancer cells, parallel to the effect of miR-132 on the lung cancer cells. Meanwhile, knockdown of ZEB2 reversed the enhanced migration and invasion mediated by anti-miR-132. These results indicate that miR-132 suppresses the migration and invasion of NSCLC cells through targeting ZEB2 involving the EMT process. Thus, our finding provides new insight into the mechanism of NSCLC progression. Therapeutically, miR-132 may serve as a potential target in the treatment of human lung cancer.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0091827PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3953608PMC
December 2015

Overexpression of CD73 in prostate cancer is associated with lymph node metastasis.

Pathol Oncol Res 2013 Oct 8;19(4):811-4. Epub 2013 May 8.

Department of Genitourinary Oncology, Tianjin Medical University Cancer Institute and Hospital, Key Laboratory of Cancer Prevention and Therapy Tianjin, Tianjin, 300060, China,

Prostate cancer is the most common malignancy in men in Europe and North America. At present, it is becoming an increasingly common cancer in China. CD73 (ecto-5'-nucleotidase) is a glycosylphosphatidylinositol (GPI)-linked 70-kDa cell surface enzyme. It is also broadly expressed in many types of tissues. Recent studies have showed that CD73 is widely expressed on malignancies and is up-regulated in cancerous tissues. Consequently, we analyzed the expression of CD73 in prostate cancer tissue. The expression of the CD73 protein was evaluated by Immunohistochemistry staining in 116 tissue specimens. The expression was further examined by quantitative real-time PCR (qRT-PCR) and Western blot in the same set of patients. The intense cell membrane staining for the CD73 protein was observed. The expression of CD73 in lymph node non-metastasizing prostate cancer tissues can be seen at low levels, and is generally undetectable. RT-PCR and Western blot showed that the expression of CD73 in lymph node metastasizing prostate cancer was higher compared with non-metastasizing ones. These results suggest that CD73 could be considered as a relevant-specific target for molecular therapy of prostate cancer metastasis.
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http://dx.doi.org/10.1007/s12253-013-9648-7DOI Listing
October 2013

[Lentivirus-mediated stable silencing of nm23-H1 gene in lung cancer cells and the influence on biological behavior].

Zhongguo Fei Ai Za Zhi 2012 Mar;15(3):139-45

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin 300052, China.

Background And Objective: The nm23-H1 gene is an important tumor metastatic suppressor gene. Our previous study showed that the downregulation of nm23-H1 gene expression using small interfering RNA (siRNA) in NL9980 lung cancer cells greatly enhanced their invasiveness. To further explore the molecular mechanisms after nm23-H1 gene knockdown, we established transgene NL9980 and A549 lung cancer cell lines with stable nm23-H1 gene silencing through the lentivirus-mediated short hairpin RNA (shRNA) method.

Methods: The human large cell lung cancer NL9980 and human lung adenocarcinoma A549 cells were transfected with shRNA lentiviral particles specific for the nm23-H1 gene, and were then selected through puromycin. Puromycin-resistant clones were generated and screened using reverse transcription polymerase chain reaction (RT-PCR), quantitative real-time polymerase chain reaction (qPCR), and Western blot analysis. shRNA rescue experiments were performed to restore the nm23-H1 gene expression in the shRNA-expressing cells. Invasiveness was determined through a Boyden chamber assay.

Results: The puromycin-resistant clones (NL9980-99 and A549-99) showed very low levels of nm23-H1 mRNA and protein expression under RT-PCR, qPCR, and Western blot analysis. Meanwhile, the shRNA rescue experiment restored the nm23-H1 expression in the NL9980-99 and A549-99 cells detected by Western blot. Downregulation of nm23-H1 gene expression enhanced the invasiveness of the NL9980-99 and A549-99 cells compared with the controls.

Conclusions: The lung cancer cell lines NL9980-99 and A549-99 with stable nm23-H1 gene silencing were successfully established and their invasiveness was greatly increased after nm23-H1 gene knockdown.
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http://dx.doi.org/10.3779/j.issn.1009-3419.2012.03.02DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5999872PMC
March 2012

[Current status and prospect of lung cancer gene therapy].

Zhongguo Fei Ai Za Zhi 2011 Sep;14(9):758-62

Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenviroment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin 300052, China.

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http://dx.doi.org/10.3779/j.issn.1009-3419.2011.09.11DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5999612PMC
September 2011