Publications by authors named "Satyajit Pradhan"

21 Publications

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Multinational Study to Assess Stress Levels Among the Health Care Workers of Radiation Oncology Community at the Outset of the COVID-19 Pandemic.

JCO Glob Oncol 2021 04;7:464-473

Department of Radiation Oncology, Tata Memorial Centre, Homi Bhabha National Institute, Mumbai, India.

Purpose: To evaluate stress levels among the health care workers (HCWs) of the radiation oncology community in Asian countries.

Methods: HCWs of the radiation oncology departments from 29 tertiary cancer care centers of Bangladesh, India, Indonesia and Nepal were studied from May 2020 to July 2020. A total of 758 eligible HCWs were identified. The 7-Item Generalized Anxiety Disorder, 9-Item Patient Health Questionnaire, and 22-Item Impact of Events Scale-Revised were used for assessing anxiety, depression, and post-traumatic stress disorder. Univariate and multivariate analysis was done to identify the causative factors affecting mental health.

Results: A total of 758 participants from 794 HCWs were analyzed. The median age was 31 years (IQR, 27-28). The incidence of moderate to severe levels of anxiety, depression, and stress was 34.8%, 31.2%, and 18.2%, respectively. Severe personal concerns were noticed by 60.9% of the staff. On multivariate analysis, the presence of commonly reported symptoms of COVID-19 during the previous 2 weeks, contact history (harzard ratio [HR], 2.04; CI, 1.15 to 3.63), and compliance with precautionary measures (HR, 1.69; CI, 1.19 to 2.45) for COVID-19 significantly predicted for increasing anxiety (HR, 2.67; CI, 1.93 to 3.70), depression (HR, 3.38; CI 2.36 to 4.84), and stress (HR, 2.89; CI, 1.88 to 4.43) ( < .001). A significant regional variation was also noticed for anxiety, stress, and personal concerns.

Conclusion: This survey conducted during the COVID-19 pandemic revealed that a significant proportion of HCWs in the radiation oncology community experiences moderate to severe levels of anxiety, depression, and stress. This trend is alarming and it is important to identify and intervene at the right time to improve the mental health of HCWs to avoid any long-term impacts.
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http://dx.doi.org/10.1200/GO.20.00647DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8081510PMC
April 2021

An assessment of serum oxidative stress and antioxidant parameters in patients undergoing treatment for cervical cancer.

Free Radic Biol Med 2021 May 9;167:29-35. Epub 2021 Mar 9.

Radiotherapy and Radiation Medicine, Banaras Hindu University, Institute of Medical Sciences, Varanasi, 221005, Uttar Pradesh, India. Electronic address:

Objectives: Oxidative stress and antioxidants are involved in all aspects of cervical cancer. The present study evaluated serum levels of oxidative stress and antioxidant biomarkers in cervical cancer patients and healthy controls. Moreover, the effect of Concurrent chemoradiotherapy (CCRT) on these biomarkers and their association with treatment outcome was investigated.

Design: This study included ninety-seven cervical cancer patients and thirty controls. Three oxidative stress parameters (8-hydroxy-2-deoxyguanosine, Protein Carbonyl, and Malondialdehyde) and four antioxidant parameters (Superoxide Dismutase, Catalase, Glutathione Peroxidase, and Total Antioxidant Status) were measured. The analysis was conducted using repeated measures ANOVA for comparing among the phases (before, during, and follow-up) of treatment. The control group was compared using the Dunnet test. Logistic regression analysis was also conducted between oxidative stress and antioxidant parameters to study their association.

Results: Significant rises in oxidative damage markers were observed in cervical cancer patients of all stages, compared to controls. There was a further increase in oxidative stress markers during CCRT among complete responders. However, among non-responders, the oxidative stress biomarkers like Protein Carbonyl and Malondialdehyde were unaltered during CCRT. Simultaneously, there was a significant decrease in antioxidant parameters in cervical cancer patients of all stages compared to controls. During CCRT, antioxidant levels continuously depleted among complete responders. Nevertheless, in non-responders, antioxidant parameters like Superoxide Dismutase and Total Antioxidant Status were consistent. The oxidative stress markers and antioxidant parameters normalized among complete responders at six months follow up. While in non-responders, the normalization of these parameters was not observed.

Conclusion: Our results indicate that increased oxidative stress and diminished antioxidants among patients were associated with carcinoma cervix. Induced oxidative stress and decreased antioxidant parameters during CCRT among the complete responders show the treatment's efficacy. Oxidant-antioxidant profile merits investigation as markers of diagnosis, treatment response, survival, and recurrence in extensive prospective studies.
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http://dx.doi.org/10.1016/j.freeradbiomed.2021.02.037DOI Listing
May 2021

Diagnostic and prognostic application of Raman spectroscopy in carcinoma cervix: A biomolecular approach.

Spectrochim Acta A Mol Biomol Spectrosc 2021 Apr 22;250:119356. Epub 2020 Dec 22.

Department of Physics, Banaras Hindu University, Institute of Science, Varanasi 221005, Uttar Pradesh, India. Electronic address:

Blood serum samples from 63 cervical cancer patients and 30 controls were collected at three different phases of the treatment (i.e. before, during, and at follow up). The spectra of serum samples from control as well as patients were classified into different groups using principal component analysis (PCA) and linear discriminant analysis (LDA) based on different phases of treatment using R software. The spectra of blood serum samples have shown the distinct changes and differences compared with each other in the profile of various biochemical parameters. The sensitivity (92.5%) and specificity (85%) were observed maximum between control and cervical cancer patients (before treatment). Between different phases of treatment, the sensitivity and specificity were less but, all accuracies of detection and classification reached above 50%. This method can be considered as a screening method for detection and treatment monitoring.
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http://dx.doi.org/10.1016/j.saa.2020.119356DOI Listing
April 2021

A comparison between revised Manchester Point A and ICRU-89-recommended Point A definition absorbed-dose reporting using CT images in intracavitary brachytherapy for patients with cervical carcinoma.

Brachytherapy 2021 Jan-Feb;20(1):118-127. Epub 2020 Aug 16.

Department of Radiotherapy & Radiation Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh, India. Electronic address:

Purpose: This study is a comparison between revised Manchester Point A and International Commission on Radiation Units and measurements (ICRU) 89 report-recommended Point A absorbed-dose reporting in intracavitary brachytherapy for patients with cervical carcinoma.

Methods And Materials: The retrospective dosimetric study is based on the data of 32 patients with cervical carcinoma treated with high-dose-rate brachytherapy. Patients received 21 Gy in three fractions (7.0 Gy X three fractions) to Point A (A, revised Manchester definition). All the patients were replanned with a new Point A (A) defined on CT images as per the American Brachytherapy Society/ICRU-89. The data collected were compared with the data obtained from Point A (A).

Results: When using the A plan normalization method, the mean dose of 0.1 cc, 1 cc, and 2 cc bladder volumes was 820.79 ± 207.47 cGy, 654.66 ± 152.69 cGy, and 588.91 ± 136.35 cGy, respectively. Likewise, when using the ICRU-89 Point A normalization method, the mean dose of 0.1 cc, 1 cc, and 2 cc bladder volumes was 869.30 ± 224.67 cGy, 693.24 ± 166.20 cGy, and 616.61 ± 150.32 cGy, respectively. For the rectum, Point A normalization plans, the mean dose of 0.1 cc, 1 cc, and 2 cc volumes was 589.37 ± 163.26 cGy, 487.51 ± 126.03 cGy, and 442.70 ± 111.43 cGy, respectively. Likewise, using the A plan, the mean 0.1 cc, 1 cc, and 2 cc rectum volume was 625.07 ± 171.31 cGy, 517.50 ± 131.05 cGy, 464.94 ± 121.81 cGy, respectively. The statistical mean difference of Total Reference Air Kerma rate, V (cc), bladder, rectum and sigmoid, was found significant.

Conclusions: It has been found that the position of revised Manchester (A) and ICRU-89 Point A does not match on CT images/radiograph, which resulted in variation in doses to the tumor, V (cc), organ at risk, and Total Reference Air Kerma.
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http://dx.doi.org/10.1016/j.brachy.2020.07.009DOI Listing
August 2020

A comparison of six fractions per week chemoradiation versus five fractions per week of conventional chemoradiation in carcinoma cervix: A prospective controlled study.

J Cancer Res Ther 2019 Oct-Dec;15(6):1296-1303

Department of Radiotherapy and Radiation Medicine, Banaras Hindu University, Varanasi, Uttar Pradesh, India.

Aims: The standard of care for carcinoma cervix stage IB2-IVA is five fractions per week of radiotherapy (RT) with concurrent cisplatin. We compared the standard treatment with six fractions per week of RT with concurrent Cisplatin to see whether the later had improved survival outcomes with comparable toxicities.

Settings And Design: 46 patients of carcinoma cervix with stage IB2-IVAwere randomized into two arms.

Materials And Methods: Study arm: 46 Gy/23 fractions/26 days, 6 fractions/week with injection CDDP 40 mg/m and Control arm: 46 Gy/23 fractions/31 days, 5 fractions/week with injection Cisplatin 40mg/m. Patients in both the arms received LDR brachytherapy to a dose of 29 Gy at point A.

Statistical Analysis Used: The primary end points were disease-free survival (DFS) and overall survival (OS). Compliance to treatment and treatment toxicities were the secondary end points. P value ≤0.05 were considered significant.

Results: The study was carried out during June, 2014-April, 2015. Statistical analysis was done in May, 2019. Of 46 patients, 39 patients completed the treatment. The study and control arms had 17 and 22 patients, respectively. Median follow-up period is 45 months (range: 1-54 months). 3-year DFS rates and OS was 69.5% vs. 72.7% (P = 0.73) and 63% vs. 68% (P = 0.45) in study and in control arm, respectively. There was no significant difference in acute and late radiation toxicities between two arms.

Conclusion: Chemoradiotherapy with six fractions per week seems feasible and equally efficacious in terms of survival outcomes and toxicity profile. Further prospective randomized controlled study is required to prove the merit of altered fractionation with concurrent cisplatin.
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http://dx.doi.org/10.4103/jcrt.JCRT_698_19DOI Listing
May 2020

Benzothiazole derivative bearing amide moiety induces p53-mediated apoptosis in HPV16 positive cervical cancer cells.

Invest New Drugs 2020 08 20;38(4):934-945. Epub 2019 Aug 20.

Department of Molecular and Human Genetics, Institute of Science, Banaras Hindu University, Varanasi, 221005, India.

In our previous study, we screened the anti-cancer properties of 10 benzothiazole derivatives in cervical cancer cell lines. In the present study, we aimed to delineate the mechanism of the apoptotic pathway (whether intrinsic or extrinsic) following the treatment of N-(4-(benzo[d]thiazol-2-yl)phenyl)-5-chloro-2-methoxybenzamide (named as A-07) on cervical cancer cell lines. Cellular stress by reactive oxygen species was measured using DCFDA dye by flowcytometry. Protein expression and localization was checked by immunofluorescence for γH2A.X, TP53, and CASP-3. Expression profiles of BAX and BCL-2 was done by semi-quantitative RT-PCR and PARP-1 (Poly(ADP-ribose) polymerase-1) by Western blot analysis. Bioinformatic studies were done using PDB websites, metaPocket 2.0 server, YASARA software and Discovery Studio 3.5 Visualizer. We demonstrate that the compound A-07 leads to ROS generation and double strand breaks in SiHa and C-33A cells. The induction of apoptosis in SiHa cells is associated with increased nuclear expression of the tumor suppressor protein, TP53. The shift in BAX/BCL-2 ratio, increased expression of Caspase-3 and cleaved Poly(ADP-ribose) polymerase-1 favour apoptotic signal in SiHa. In silico studies revealed that A-07 has inhibiting capabilities to the E6/E6AP/P53 complex. Our data suggest that treatment of A-07 causes p53 and caspase dependent apoptosis in HPV 16 infected SiHa cells.
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http://dx.doi.org/10.1007/s10637-019-00848-7DOI Listing
August 2020

Search of an ideal location of isocenter in intensity-modulated radiotherapy treatment plans: A dosimetrical approach.

J Cancer Res Ther 2019 Jan-Mar;15(1):211-215

Department of Radiotherapy and Radiation Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh, India.

Aim: The aim of this study is to identify an ideal location of isocenter in intensity-modulated radiotherapy (IMRT) treatment plans.

Materials And Methods: A total of 28 clinical target volumes and 4 English capital letters (C, L, T, and H) target volumes were considered in this study. Two IMRT treatment plans were generated for each target volume in the ECLIPSE treatment planning system (TPS), first one with isocenter automatically placed (ISO) by TPS and the second one with geometric center-based isocenter (ISO). The geometric center of a cuboid volume, which was formed encompassing around the target volume in sagittal, transverse, and frontal planes, is considered as the geometric center of the target volume as well as the isocenter (ISO) of the IMRT plans. While performing the IMRT treatment plans using the beam angle optimization and dose volume optimization, the normal tissue objectives and target volume objectives were kept similar in both the plans. The dosimetrical parameters between the two groups of plans were compared.

Results: The distance between ISO and ISO ranged from 0.16 cm to 3.04 cm with a mean and median of 0.85 cm and 0.69 cm, respectively. The ISO-based IMRT plans exhibited statistically significant advantages in total monitor units reduction (100% of cases, P ≤ 0.001), total number of field reduction (66% of cases, P ≤ 0.001), and reduction of patient mean dose (69% of cases, P ≤ 0.001) over ISO-based IMRT plans. The conformity index, homogeneity index and target mean dose were comparable between both group of plans.

Conclusion: Significant dosimetrical advantages may be observed, when the geometric centroid of target volume is considered as isocenter of IMRT treatment plan.
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http://dx.doi.org/10.4103/jcrt.JCRT_985_16DOI Listing
July 2019

Olaparib modulates DNA repair efficiency, sensitizes cervical cancer cells to cisplatin and exhibits anti-metastatic property.

Sci Rep 2017 10 9;7(1):12876. Epub 2017 Oct 9.

Department of Molecular and Human Genetics, Institute of Science, Banaras Hindu University, Varanasi, India.

PARP1 trapping at DNA lesion by pharmacological inhibitors has been exploited in several cancers exhibiting defects in DNA repair mechanisms. PARP1 hyperactivation is involved in therapeutic resistance in multiple cancers. The role of PARP1 in cervical cancer (CC) resistance and implication of PARP inhibitor is yet to be elucidated. Our data demonstrates significantly higher expression of PARP1 in primary cervical tumors and CC cell lines SiHa and ME180. Upon cisplatin treatment CC cells display significant overexpression of PARP1 and its hyperactivation. PARP inhibitor olaparib shows significant anti-proliferative effect on CC cells and drive loss of clonogenic survival and enhanced cell death in combination with cisplatin. PARP inhibited cells show delay in resolution of γH2A.X foci and prolonged late S and G2-M phase arrest resulting in apoptosis. Further, PARP inhibition disrupts the localization of base excision repair (BER) effector XRCC1 and non-homologous end joining (NHEJ) proteins Ku80 and XRCC4. Due to disrupted relocation of repair factors, cisplatin induced stalled replication forks collapse and convert into double strand breaks (DSBs). Interestingly, PARP inhibition also shows anti-migratory and anti-invasive properties in CC cells, increases anchorage independent cell death and induces anoikis. Collectively, our data demonstrates therapeutic potential of PARP inhibitor in cervical cancer.
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http://dx.doi.org/10.1038/s41598-017-13232-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5634505PMC
October 2017

CXCL12 is a key regulator in tumor microenvironment of cervical cancer: an in vitro study.

Clin Exp Metastasis 2016 06 12;33(5):431-9. Epub 2016 Mar 12.

Cancer Genetics Laboratory, Department of Molecular and Human Genetics, Banaras Hindu University, Varanasi, 221005, India.

CXCL12 is a small pro-inflammatory chemo-attractant cytokine which signals through chemokine receptor CXCR4. The importance of CXCL12/CXCR4 axis is coming to the fore in several divergent signaling pathway-initiating signals related to cell survival and/or proliferation and cancer metastasis. In the present study we have investigated whether deregulation in CXCR4 signaling (as a consequence of deregulated expression of CXCL12) modulate the metastatic potential of cervical carcinoma cells. We demonstrate that CXCL12 is frequently down regulated and its promoter is hypermethylated in cervical cancer cell lines and primary tumor biopsies. Exogenous treatment of cervical cancer cell lines (HeLa, SiHa and C-33A) with recombinant CXCL12 inhibited the metastasis promoting cell migration, cell invasion and anchorage independent cell growth events. Although this study will need further in vivo validation, our observations suggest that (a) silencing of CXCL12 in cervical cancer cells may be critical in migration and invasion, the key events in cancer cell metastases; (b) cervical cancer cells having down regulated CXCL12 are more prone to being attracted to CXCL12 expressed at secondary sites of metastases; and (c) CXCL12 inhibits anchorage independent cell growth via anoikis. These findings suggest the tumor suppressor functions of CXCL12 in cervical cancer.
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http://dx.doi.org/10.1007/s10585-016-9787-9DOI Listing
June 2016

HPV-type-specific response of cervical cancer cells to cisplatin after silencing replication licensing factor MCM4.

Tumour Biol 2015 Dec 19;36(12):9987-94. Epub 2015 Jul 19.

Cancer Genetics Laboratory, Department of Molecular and Human Genetics, Banaras Hindu University, Varanasi, 221 005, Uttar Pradesh, India.

Minichoromosome maintenance (MCM) proteins play key role in cell cycle progression by licensing DNA replication only once per cell cycle. These proteins are found to be overexpressed in cervical cancer cells. In this study, we depleted MCM4, one of the MCM 2-7 complex components by RNA interference (RNAi) in four cervical cancer cell lines. The four cell lines were selected on the basis of their human papillomavirus (HPV) infection: HPV16-positive SiHa, HPV18-positive ME-180, HPV16- and HPV18-positive CaSki, and HPV-negative C-33A. The MCM4-deficient cells irrespective of their HPV status grow for several generations and maintain regular cell cycle. We did not find any evidence of augmented response to a short-term (48 h) cisplatin treatment in these MCM4-deficient cells. However, MCM4-/HPV16+ SiHa cells cannot withstand a prolonged treatment (up to 5 days) of even a sublethal dosage of cisplatin. They show increased chromosomal instability compared to their control counterparts. On the other hand, MCM4-deficient CaSki cells (both HPV16+ and 18+) remain resistant to a prolonged exposure to cisplatin. Our study indicates that cervical cancer cells may be using excess MCMs as a backup for replicative stress; however, its regulatory mechanism is dependent on the HPV status of the cells.
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http://dx.doi.org/10.1007/s13277-015-3782-7DOI Listing
December 2015

Anti-tumor activity of staurosporine in the tumor microenvironment of cervical cancer: An in vitro study.

Life Sci 2015 Jul 23;133:21-8. Epub 2015 May 23.

Department of Molecular and Human Genetics, Banaras Hindu University, Varanasi 221005, India. Electronic address:

Aim: The fundamental events for cancer progression and metastases include loss of cell adhesion, cell proliferation, anchorage-independent cell growth (evading anoikis), cell migration and cell invasion. All these events leading to cancer progression happen in a favorable nurturing tumor microenvironment. This study was designed to explore the anti-tumor activity of staurosporine (a nonspecific protein kinase inhibitor) in the tumor microenvironment of cervical cancer.

Main Methods: The anti-tumor activity of staurosporine was investigated by cell adhesion assay, colony formation assay, apoptosis assay and quantitative real-time polymerase chain reaction (PCR) in cervical cancer cell lines.

Key Findings: The cell adhesion assay showed that staurosporine induces adhesion of cervical cancer cells to the extracellular matrix (ECM) protein fibronectin. The soft agar colony formation assay showed that staurosporine inhibits both the number and size of colony formation in a dose dependent manner and also induces adherent tendency in the cancer cells. Staurosporine also induces prominent apoptosis in single cell suspensions compared to adherent cells. Stroma cell induced transcription of matrix metalloprotease 1 (MMP1) and matrix metalloprotease 2 (MMP2) in cervical cancer cells was inhibited by staurosporine.

Significance: Our results indicate that staurosporine induces anti-tumor response in the cervical tumor microenvironment by inhibiting the fundamental events for cancer progression and metastases. The present study represents an attractive area for further research and opens up new avenues towards the understanding of cervical cancer therapeutics.
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http://dx.doi.org/10.1016/j.lfs.2015.04.019DOI Listing
July 2015

PI3K/AKT pathway-mediated regulation of p27(Kip1) is associated with cell cycle arrest and apoptosis in cervical cancer.

Cell Oncol (Dordr) 2015 Jun 28;38(3):215-25. Epub 2015 Mar 28.

Cancer Genetics Laboratory, Department of Molecular and Human Genetics, Banaras Hindu University, Varanasi, 221005, India.

Background: The cyclin-dependent kinase inhibitor p27(Kip1) is known to act as a putative tumor suppressor in several human cancers, including cervical cancer. Down-regulation of p27(Kip1) may occur either through transcription inhibition or through phosphorylation-dependent proteolytic degradation. As yet, the mechanism underlying p27(Kip1) down-regulation and its putative downstream effects on cervical cancer development are poorly understood. Here we assessed the expression and sub-cellular localization of p27(Kip1) and its effects on proliferation, cell cycle progression and (inhibition of) apoptosis in cervical cancer cells.

Methods: Primary cervical cancer samples (n = 70), normal cervical tissue samples (n = 30) and cervical cancer-derived cell lines (n = 8) were used to assess the expression of p27(Kip1) and AKT1 by RT-PCR, Western blotting and immunohistochemistry, respectively. The effects of the PI3K inhibitor LY294004 and the proteasome inhibitor MG132 on cervical cancer cell proliferation were investigated using a MTT assay. Apoptosis and cell cycle analyses were carried out using flow cytometry, and sub-cellular p27(Kip1) localization analyses were carried out using immunofluorescence assays.

Results: We observed p27(Kip1) down-regulation (p = 0.045) and AKT1 up-regulation (p = 0.046) in both the primary cervical cancer samples and the cervical cancer-derived cell lines, compared to the normal cervical tissue samples tested. Treatment of cervical cancer-derived cell lines with the PI3K inhibitor LY294002 resulted in a reduced AKT1 activity. We also observed a dose-dependent inhibition of cell viability after treatment of these cell lines with the proteasome inhibitor MG132. Treatment of the cells with LY294002 resulted in a G1 cell cycle arrest, a nuclear expression of p27(Kip1), and a cytoplasmic p27(Kip1) accumulation after subsequent treatment with MG132. Additionally, we found that the synergistic effect of MG132 and LY294002 resulted in a sub-G1 cell cycle arrest and apoptosis induction through poly (ADP-ribose) polymerase (PARP) cleavage.

Conclusion: Our data suggest that p27(Kip1) down-regulation in cervical cancer cells is primarily regulated through PI3K/AKT-mediated proteasomal degradation. The observed synergistic effect of the MG132 and LY294002 inhibitors may form a basis for the design of novel cervical cancer therapies.
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http://dx.doi.org/10.1007/s13402-015-0224-xDOI Listing
June 2015

MCM Paradox: Abundance of Eukaryotic Replicative Helicases and Genomic Integrity.

Mol Biol Int 2014 19;2014:574850. Epub 2014 Oct 19.

Cancer Genetics Laboratory, Department of Molecular and Human Genetics, Banaras Hindu University, Varanasi, India.

As a crucial component of DNA replication licensing system, minichromosome maintenance (MCM) 2-7 complex acts as the eukaryotic DNA replicative helicase. The six related MCM proteins form a heterohexamer and bind with ORC, CDC6, and Cdt1 to form the prereplication complex. Although the MCMs are well known as replicative helicases, their overabundance and distribution patterns on chromatin present a paradox called the "MCM paradox." Several approaches had been taken to solve the MCM paradox and describe the purpose of excess MCMs distributed beyond the replication origins. Alternative functions of these MCMs rather than a helicase had also been proposed. This review focuses on several models and concepts generated to solve the MCM paradox coinciding with their helicase function and provides insight into the concept that excess MCMs are meant for licensing dormant origins as a backup during replication stress. Finally, we extend our view towards the effect of alteration of MCM level. Though an excess MCM constituent is needed for normal cells to withstand stress, there must be a delineation of the threshold level in normal and malignant cells. This review also outlooks the future prospects to better understand the MCM biology.
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http://dx.doi.org/10.1155/2014/574850DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4217321PMC
November 2014

Down Regulation of FOXO1 Promotes Cell Proliferation in Cervical Cancer.

J Cancer 2014 22;5(8):655-62. Epub 2014 Aug 22.

1. Cancer Genetics Laboratory, Department of Molecular and Human Genetics, Banaras Hindu University, Varanasi-221 005, India;

The Forkhead transcription factor FOXO1, an important downstream target of phosphatidylinositol-3-kinase (PI3K)/AKT signaling pathway, regulates cellular homeostasis by maintaining cell proliferation, apoptosis and viability in normal cells. Though, the function and regulation of FOXO1 is well documented in many cancers, the molecular mechanism of its regulation in cervical cancer is largely unknown. In the present study we have investigated the role of PI3K inhibition on FOXO1 regulation. Expression profiling of primary tumors and cell lines show over expression of PIK3CA and AKT1; and down regulation of FOXO1. Lack of FOXO1 promoter methylation and inability of hypomethylating drug 5-Aza-2'-deoxycytidine and HDAC inhibitor trichostatin A to reactivate FOXO1 expression suggest that loss of FOXO1 expression is due to mechanisms other than promoter methylation/acetylation. Inhibition of PI3K by LY294002 decreased the level of p-AKT1 and activated FOXO1 transcription factor. We demonstrate that activation of FOXO1 induces apoptosis, cell proliferation arrest, and decreased cell viability in cervical cancer cell lines. Our data suggest that frequent down regulation of FOXO1 and its functional inactivation may be due to post-translational modifications in cervical cancer. Together, these observations suggest that activation of FOXO1 and its nuclear sequestration is critical in the regulation of cell proliferation, cell viability and apoptosis in cervical cancer. Hence, PI3K/AKT pathway may be a potential molecular target for cervical cancer therapy.
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http://dx.doi.org/10.7150/jca.6554DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4142327PMC
August 2014

Epigenetic silencing of CXCR4 promotes loss of cell adhesion in cervical cancer.

Biomed Res Int 2014 10;2014:581403. Epub 2014 Jul 10.

Department of Molecular and Human Genetics, Banaras Hindu University, Varanasi 221005, India.

In the network of chemokine signaling pathways, recent reports have described the SDF-1α/CXCR4 axis and its role in cancer progression and metastasis. Interestingly, we found downregulation of CXCR4 at both transcript and protein level in cervical cancer cell lines and primary tumors. We also found CXCR4 promoter hypermethylation in cervical cancer cell lines and primary biopsy samples. DNA hypomethylating drug 5-AZA-2'-deoxycytidine and histone deacetylase inhibitor Trichostatin A treatments in cell lines reactivate both CXCR4 transcription and protein expression. Cell adhesion assay demonstrated that autocrine SDF-1α promotes the loss of cell adhesion while paracrine SDF-1α predominantly protects the normal cervical cells from loss of cell adhesion. Cervical cancer cell line C-33A having increased expression of CXCR4 after TSA treatment showed increased cell adhesion by paracrine source of SDF-1α in comparison to untreated C-33A. These findings demonstrate the first evidence that epigenetic silencing of CXCR4 makes the cells inefficient to respond to the paracrine source of SDF-1α leading to loss of cell adhesion, one of the key events in metastases and progression of the disease. Our results provide novel insight of SDF-1α/CXCR4 signaling in tumor microenvironment which may be promising to further delineate molecular mechanism of cervical carcinogenesis.
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http://dx.doi.org/10.1155/2014/581403DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4119908PMC
April 2015

Over expression of minichromosome maintenance genes is clinically correlated to cervical carcinogenesis.

PLoS One 2013 17;8(7):e69607. Epub 2013 Jul 17.

Cancer Genetics Laboratory, Department of Molecular and Human Genetics, Banaras Hindu University, Varanasi, Uttar Pradesh, India.

Minichromosome Maintenance (MCM) proteins play important roles in cell cycle progression by mediating DNA replication initiation and elongation. Among 10 MCM homologues MCM 2-7 form a hexamer and assemble to the pre-replication complex acting as replication licensing factors. Binding and function of MCM2-7 to pre-replication complex is regulated by MCM10 mediated binding of RECQL4 with MCM2-7. The purpose of this study is to explore the role of MCMs in cervical cancer and their correlation with the clinical parameters of cervical cancer. We have investigated sixty primary cervical cancer tissue samples, eight cervical cancer cell lines and thirty hysterectomised normal cervical tissue. The expression profiling of MCMs was done using semi-quantitative RT-PCR, immunoblotting and immunohistochemistry. MCM2, 4, 5, 6, 7, 10 and RECQL4 are significantly over-expressed in cervical cancer. Among these, MCM4, 6 and 10 show increased frequency of over expression along with advancement of tumor stages. MCM4, 5 and 6 also show differential expression in different types of lesion, while MCM2 and MCM10 are over expressed in cervical cancer irrespective of clinico-pathological parameters. Our data indicates the role of MCM4, MCM5, MCM6, MCM10 and RECQL4 in the progression of cervical cancer.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0069607PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3714251PMC
March 2014

Assessment of predictive markers of response to neoadjuvant chemotherapy in breast cancer.

Asian J Surg 2010 Oct;33(4):157-67

Department of Surgical Oncology, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India.

Objective: To identify the predictive markers associated with chemotherapy sensitivity, especially those producing pathological complete response (pCR) following neoadjuvant chemotherapy (NACT) in patients with locally advanced breast cancer.

Methods: Core needle biopsy of 50 locally advanced breast cancer patients was analysed for histopathology, grade, oestrogen receptor, progesterone receptor, HER2, Ki-67, p53, Bcl-2, and BAX before starting NACT. This was correlated with response to NACT using Response Evaluation Criteria in Solid Tumours criteria.

Results: The mean tumour reduction rate per chemotherapy cycle was significantly higher in BAX-positive (p = 0.01) and Bcl-2-negative (p = 0.04) tumours. BAX expression significantly (p = 0.043) correlated with a response of an at least 30% reduction in tumour size post-NACT on multivariate analysis. A significant relationship was seen between loss of Bcl-2 expression and pCR on univariate (p = 0.048) analysis. Overall, all of the above 12 parameters had 30.4% and 28.5% success in predicting clinical complete response and pCR, respectively, by the Cox and Snell formula.

Conclusion: Of all parameters examined, only the apoptosis-related genes (Bcl-2 and BAX) seemed to exert some influence on the response to NACT, and neither by itself was sufficient to predict pCR; however, 50 patients is not sufficient to simultaneously analyse several predictive markers.
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http://dx.doi.org/10.1016/S1015-9584(11)60001-8DOI Listing
October 2010

Segmentation and classification of medical images using texture-primitive features: Application of BAM-type artificial neural network.

J Med Phys 2008 Jul;33(3):119-26

School of Biomedical Engineering, Institute of Technology, Banaras Hindu University, Varanasi (UP), India.

The objective of developing this software is to achieve auto-segmentation and tissue characterization. Therefore, the present algorithm has been designed and developed for analysis of medical images based on hybridization of syntactic and statistical approaches, using artificial neural network (ANN). This algorithm performs segmentation and classification as is done in human vision system, which recognizes objects; perceives depth; identifies different textures, curved surfaces, or a surface inclination by texture information and brightness. The analysis of medical image is directly based on four steps: 1) image filtering, 2) segmentation, 3) feature extraction, and 4) analysis of extracted features by pattern recognition system or classifier. In this paper, an attempt has been made to present an approach for soft tissue characterization utilizing texture-primitive features with ANN as segmentation and classifier tool. The present approach directly combines second, third, and fourth steps into one algorithm. This is a semisupervised approach in which supervision is involved only at the level of defining texture-primitive cell; afterwards, algorithm itself scans the whole image and performs the segmentation and classification in unsupervised mode. The algorithm was first tested on Markov textures, and the success rate achieved in classification was 100%; further, the algorithm was able to give results on the test images impregnated with distorted Markov texture cell. In addition to this, the output also indicated the level of distortion in distorted Markov texture cell as compared to standard Markov texture cell. Finally, algorithm was applied to selected medical images for segmentation and classification. Results were in agreement with those with manual segmentation and were clinically correlated.
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http://dx.doi.org/10.4103/0971-6203.42763DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2772042PMC
July 2008

Doppler ultrasound scoring to predict chemotherapeutic response in advanced breast cancer.

World J Surg Oncol 2007 Aug 28;5:99. Epub 2007 Aug 28.

Department of Surgery, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, India.

Background: Doppler ultrasonography (US) is increasingly being utilized as an imaging modality in breast cancer. It is used to study the vascular characteristics of the tumor. Neoadjuvant chemotherapy is the standard modality of treatment in locally advanced breast cancer. Histological examination remains the gold standard to assess the chemotherapy response. However, based on the color Doppler findings, a new scoring system that could predict histological response following chemotherapy is proposed.

Methods: Fifty cases of locally advanced infiltrating duct carcinoma of the breast were studied. The mean age of the patients was 44.5 years. All patients underwent clinical, Doppler and histopathological assessment followed by three cycles of CAF (Cyclophosphamide, Adriamycin and 5-Fluorouracil) chemotherapy, repeat clinical and Doppler examination and surgery. The resected specimens were examined histopathologically and histological response was correlated with Doppler findings. The Doppler characteristics of the tumor were graded as 1-4 for <25%, 25-50%, >50% and complete disappearance of flow signals respectively. A cumulative score was calculated and compared with histopathological response. Results were analyzed using Chi square test, sensitivity, specificity, positive and negative predictive values.

Results: The maximum Doppler score according to the proposed scoring system was twelve and minimum three. Higher scores corresponded with a more favorable histopathological response. Twenty four patients had complete response to chemotherapy. Sixteen of these 24 patients (66.7%) had a cumulative Doppler score more than nine. The sensitivity of cumulative score >5 was 91.7% and specificity was 38.5%. The area under the ROC curve of the cumulative score >9 was 0.72.

Conclusion: Doppler scoring can be accurately used to objectively predict the response to chemotherapy in patients with locally advanced breast cancer and it correlates well with histopathological response.
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http://dx.doi.org/10.1186/1477-7819-5-99DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2008196PMC
August 2007

Radiation induced meningioma with a short latent period following high dose cranial irradiation - case report and literature review.

J Neurooncol 2006 Mar 15;77(1):73-7. Epub 2005 Nov 15.

Department of Neurosurgery, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221 005, Uttar Pradesh, India.

Radiation induced meningiomas (RIM) are rare late complications in patients who have received high dose irradiation for brain tumors. The mean latency period for induction of RIM in most of the series is 18.7+/-10.2 years. There are only 9 reported cases of RIM following high dose cranial irradiation with unusually short latency periods of less than 5 years. Herein, we report a child diagnosed with RIM with an unusually short latency period of 14 months. An 11-year old male child underwent gross total resection of medulloblastoma. Following surgery he received high dose craniospinal irradiation. Postoperative computed tomography scan (CT scan) after 1 month did not show features of any residual tumor, recurrence or tumor at a new site. The child was asymptomatic for 14 months and then presented with complaints of headache and vomiting. CT scan head showed multiple solid homogenously enhancing lesions in bilateral basifrontal and right basitemporal region. Histopathology of the lesions turned out to be atypical meningioma.
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http://dx.doi.org/10.1007/s11060-005-9009-9DOI Listing
March 2006

Color Doppler ultrasound as an objective assessment tool for chemotherapeutic response in advanced breast cancer.

Breast Cancer 2005 ;12(1):45-51

Department of General Surgery, Institute of Medical Sciences, B-4 New Medical enclave Banaras Hindu University Varanasi-221005, India.

Background: In our part of the world, the majority of the patients with breast cancer present with locally advanced disease and require neo-adjuvant chemotherapy as the primary treatment modality. It is essential to monitor the response to chemotherapy in these patients. Clinical examination as the sole criterion of response assessment is entirely subjective and fallacious. Magnetic Resonance Imaging (MRI) and Computed Tomography (CT) are expensive. The role of Doppler ultrasonography as an imaging modality for this purpose is therefore being evaluated.

Methods: A prospective study was undertaken of 25 cases of locally advanced breast carcinoma (LABC) and Color Doppler sonography was used for the sequential assessment of chemotherapeutic response. The response assessed on the basis of clinical examination and Color Doppler was compared with the histological response. The parameters assessed on color Doppler were a change in the number of flow signals, maximum flow velocity (Vmax), pulsatility index (PI) and resistivity index (RI). Responses were analysed statistically using the Pearson correlation coefficient and Kappa statistics (kappa). The sensitivity, specificity, positive predictive & negative predictive values for predicting complete histological response were calculated.

Results: Color Doppler showed a sensitivity of 88.88 % for predicting complete histological response. The negative predictive value of color Doppler was 92.3 %. A significant correlation was obtained between color Doppler and histopathological response.

Conclusions: Color Doppler was found to be an objective and effective tool or modality compared with clinical evaluation in sequential response assessment, especially for predicting complete histological response.
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http://dx.doi.org/10.2325/jbcs.12.45DOI Listing
June 2005