Study of tumor transglutaminase 2 expression in gallbladder cancer – Is it a novel predictor of survival?

Sameer Gupta,Sudeep Garg,Vijay Kumar,Arun Chaturvedi,Sanjeev Misra,Naseem Akhtar,Shiv Rajan,Jatinder Kaur,Manikandan Lakshmanan,Kavitha Jain 한국간담췌외과학회 2020 Annals of hepato-biliary-pancreatic surgery Vol.24 No.4

Backgrounds/Aims: Transglutaminase 2 (TG2) is known to be an important mediator of inflammation induced carcinogenesis pathway. Chronic inflammation is the most important causative factor in Gallbladder cancer (GBC) carcinogenesis. We analyzed the expression of TG2 in GBC and its role as potential prognostic marker, first of its kind study. Methods: We analyzed TG2 expression in 100 cases of GBC and 28 cases of non-cancer gallbladder specimen (calculus cholecystitis). We studied TG2 expression in GBC in comparison to control group and evaluated its role as a potential prognostic marker. Results: TG2 score (1-9) was calculated by multiplying percentage cytoplasmic expression (P) with intensity of expression (I) in tumor cells. Positive TG-2 expression was observed in 62% of GBC patients compared to only 21% (n=6) in control group (p=0.001). In curative resection subgroup (n=54), TG2 positive patients showed shorter disease free survival rate (p=0.04) and higher rate of recurrence (p=0.03) compared to TG2 negative patients. TG2 positive expression was observed in 15/16 of patients with recurrent disease. In palliative treatment subgroup, patients with strong TG2 positive expression had poorer disease specific survival (p=0.01) as compared to weakly positive group. On multivariate analysis, lymph node status (p=0.03) and TG2 expression (p=0.037), were found to be significant predictor of recurrence and eventual survival. Conclusions: Positive TG2 expression was related to higher recurrence rates post curative surgery, shorter disease free and overall survival and ultimately portended poor prognosis. It may be helpful in better prognostication and tailoring therapeutic approach for better management of GBC.

Expression of DNA Methyltransferases in Breast Cancer Patients and to Analyze the Effect of Natural Compounds on DNA Methyltransferases and Associated Proteins

Sameer Mirza,Gayatri Sharma,Rajinder Parshad,Sidhartha Datta Gupta,Pranav Pandya,Ranju Ralhan 한국유방암학회 2013 Journal of breast cancer Vol.16 No.1

Purpose: The DNA methylation mediated by specific DNA methyltransferases (DNMTs), results in the epigenetic silencing of multiple genes which are implicated in human breast cancer. We hypothesized that the natural compounds modulate the expression of DNMTs and their associated proteins in the breast cancer cell lines and affect the methylation mediated gene silencing. Methods: The DNMTs transcript expression was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) in the tumors and the adjacent normal breast tissues of the patients with invasive ductal breast carcinoma. We tested the hypothesis that the natural compounds, viz., epigallocatechin gallate (EGCG), genistein, withaferin A, curcumin, resveratrol, and guggulsterone, have demethylation potential. To investigate this hypothesis, we analyzed the DNMTs expression at the transcript levels, followed by the analysis of DNMT1 and its associated proteins (HDAC1, MeCP2, and MBD2). Results: The increased DNMTs transcripts expression, viz., DNMT1, DNMT3a, and DNMT3b, in the breast cancer tissues suggest involvement of the DNMTs in the breast carcinogenesis. Quantitative RT-PCR analysis revealed that the treatment with natural compounds, viz., EGCG, genistein, withaferin A, curcumin, resveratrol, and guggulsterone, resulted in a significant decrease in the transcript levels of all the DNMTs investigated. Importantly, these natural compounds decreased the protein levels of DNMT1, HDAC1, and MeCP2. Conclusion: Our results demonstrate that the natural compounds, EGCG, genistein, withaferin A, curcumin, resveratrol, and guggulsterone, have the potential to reverse the epigenetic changes. Moreover, their lack of toxicity makes these natural compounds promising candidates for the chemoprevention of the breast cancer. In-depth future mechanistic studies aimed to elucidate how these compounds affect the gene transcription are warranted.

Diagnostic performance of microRNA- 34a, let-7f and microRNA-31 in epithelial ovarian cancer prediction

Vivek Kumar,Sameer Gupta,Kachnar Varma,Amrita Chaurasia,Manisha Sachan 대한부인종양학회 2022 Journal of Gynecologic Oncology Vol.33 No.4

Objective: To correlate the genome-wide methylation signature of microRNA genes with dysregulated expression of selected candidate microRNA in tissue and serum samples of epithelial ovarian cancer (EOC) and control using quantitative reverse transcription polymerase chain reaction (qRT-PCR), and evaluation of EOC predictive value of candidate microRNA at an early stage. Methods: We performed Methylated DNA Immunoprecipitation coupled with NGS (MeDIP- NGS) sequencing of 6 EOC and 2 normal tissue samples of the ovary. Expression of selected microRNA from tissue (EOC=85, normal=30) and serum (EOC=50, normal=15) samples was evaluated using qRT-PCR. We conducted bioinformatics analysis to identify the candidate miRNA’s potential target and functional role. Results: MeDIP-NGS sequencing revealed hypermethylation of several microRNAsgene promoters. Three candidate microRNAs were selected (microRNA-34a, let-7f, and microRNA-31) from MeDIP-NGS data analysis based on log2FC and P-value. The relative expression level of microRNA-34a, let-7f, and microRNA-31 was found to be significantly reduced in early-stage EOC tissues and serum samples (p<0.0001). The receiver operating characteristic analysis of microRNA-34a, let-7f and miR-31 showed improved diagnostic value with area under curve(AUC) of 92.0 (p<0.0001), 87.9 (p<0.0001), and 85.6 (p<0.0001) and AUC of 82.7 (p<0.0001), 82.0 (p<0.0001), and 81.0 (p<0.0001) in stage III-IV and stage I-II EOC serum samples respectively. The integrated diagnostic performance of microRNA panel (microRNA-34a+let-7f+microRNA-31) in late-stage and early-stage serum samples was 95.5 and 96.9 respectively. Conclusion: Our data correlated hypermethylation-associated downregulation of microRNA in EOC. In addition, a combined microRNA panel from serum could predict the risk of EOC with greater AUC, sensitivity, and specificity.

Intra-Operative Frozen Sections: Experience at A Tertiary Care Centre

Preeti, Agarwal,Sameer, Gupta,Kulranjan, Singh,Abhinav, Sonkar Arun,Preeti, Rani,Sunita, Yadav,Mati, Goel Madhu Asian Pacific Journal of Cancer Prevention 2016 Asian Pacific journal of cancer prevention Vol.17 No.12

The present study was conducted to assess error rates with diagnosis using intra-operative frozen sections, and to indicate ways to increase overall performance. Over a period of two years, 227 cases were biopsied intra-operatively. Errors were observed in 14 cases. Four of these were sampling errors, one by a pathologist and three by surgeons. In seven cases incorrect interpretations were made. Epithelial dysplasia was observed on definitive histology in two cases which was not reported intra-operatively. One case was of ectopic thyroid. In cases of oral cancer where sentinel lymph nodes were sampled, immunohistochemistry for cytokeratin was performed to facilitate identification of micrometastasis. Only single case displayed tumor deposits which was not evident morphologically. Resection margins were reported in seventy eight cases. Some 18% (14/50) benefited from revision of margins; overall sensitivity of intra-operative frozen sections for marginal status was 71.4%, with a specificity of 90.3%. Overall sensitivity was 75% and specificity was 97.5%. Careful observation, pathologist experience and knowledge of limitations help in improving the overall diagnostic outcome.

Promoter Methylation of PTEN Is a Significant Prognostic Factor in Melanoma Survival

Roh, Mi Ryung,Gupta, Sameer,Park, Kyu-Hyun,Chung, Kee Yang,Lauss, Martin,Flaherty, Keith T.,Jö,nsson, Gö,ran,Rha, Sun Young,Tsao, Hensin Elsevier 2016 The Journal of investigative dermatology Vol.136 No.5

<P>Structural compromise of the tumor suppressor gene, phosphatase and tensin homolog (PTEN), occurs in 10% of melanoma specimens, and loss of PTEN expression through DNA methylation of the PTEN promoter region has also been reported in a number of other malignancies. However, the role of PTEN promoter methylation in melanoma is not well understood. We thus sought to elucidate the prevalence of PTEN promoter methylation in melanoma specimens, its relationship to clinical features, and its impact on the outcome of patients with melanoma. PTEN promoter methylation data were acquired from an archived primary Korean melanoma cohort (KMC) of 158 patients and, for validation, 234 patients from The Cancer Genome Atlas melanoma (TCGA-MEL) cohort. Hierarchical clustering was performed to identify PTEN 'high methylated' and 'low methylated' samples. Subsequently, differences in clinical features and outcomes based on PTEN promoter methylation status were then analyzed using SPSS and R. In the KMC, all tumors were acquired from primary tumors and 65.7% (n = 105) were acral or mucosal by site, whereas in the TCGA-MEL cohort, 90.5% of the tumors were from regional lymph node and distant metastatic lesions. Overall, 17.7% and 45.7% of the specimens harbored BRAF mutations in the KMC and TCGA-MEL cohort, respectively. Neuroblastoma RAS viral oncogene homolog was mutated in 12.2% and 26.9% of the tumors in the KMC and TCGA-MEL cohort, respectively. In the KMC, 31 cases (19.6%) were included in the high methylated group versus 142 cases (60.7%) in the TCGA-MEL cohort (P < 0.001). Multivariate Cox-regression analysis revealed promoter methylation of PTEN to be an independent negative prognostic factor for survival in both the KMC (hazard ratio 3.76, 95% confidence interval = 1.24-11.12, P = 0.017) and TCGA-MEL cohort (HR 1.88, 95% confidence interval = 1.13-3.12, P = 0.015). Our results indicate that PTEN promoter methylation is an independent predictor for impaired survival in patients with melanoma.</P>

Effectiveness of online versus in-person structured training program on arterial blood gas, electrolytes, and ventilatory management of critically ill patients

Gaurav Jain,Bhavna Gupta,Priyanka Gupta,Sagarika Panda,Sameer Sharma,Shalinee Rao 대한중환자의학회 2021 Acute and Critical Care Vol.36 No.1

Background: Due to the risk of viral transmission during in-person training, a shift towardonline platforms is imperative in the current pandemic. Therefore, we compared the effectivenessof an in-person interactive course with a structurally similar online course designedto improve cognitive skills among clinical health professionals in arterial blood gas analysis,management of electrolyte imbalances, and approaches to mechanical ventilation in criticallyill patients. Methods: In an observational, outcome assessor-blinded, cohort trial, group A included participantsenrolled prospectively in an online course, while group B included those who tookpart in an in-person course (retrospective arm). The primary objective was comparison ofcognitive skills through a pre and post-test questionnaire. Statistical analysis was performedusing Student t-test. Results: In total, 435 participants were analyzed in group A, while 99 participants were evaluatedin group B. The mean pre-test score was 9.48±2.75 and 10.76±2.42, while the meanpost-test score was 11.94±1.90 (passing rate, 64.6%) and 12.53±1.63 (passing rate, 73.3%)in groups A and B, respectively. Group B scored significantly higher in both pre-test (P=0.001)and post-test evaluations (P=0.004). The improvement in post-test score was significantlygreater (P=0.001) in group A (2.46±2.22) compared to group B (1.77±1.76). The medicalspecialties fared better in group B, while surgical specialties scored higher in group A. Thepre-test vs. post-test scores exhibited a moderate correlation in both groups (P<0.001). Thefeedback survey showed a Likert score >3.5 for most points in both groups. Conclusions: The online teaching module exhibited a significant benefit in terms of participantsensitization and knowledge sharing.