Abstract P2-18-02: Promoter hypomethylation associated upregulation of MEN1 mRNA expression and its correlation with pathological parameters in Indian breast cancer patients

Abstract

Abstract Background: Breast cancer in the recent years has accounted to be most frequently reported cancer across the globe. Nearly 11.7% of total cancer cases were of breast cancer and 6,84,996 deaths were reported in the year 2020. Multiple endocrine neoplasia type 1 gene that encodes for menin protein is known be involved in numerous signaling pathways and is also reported to show its interactions with histone modifiers. MEN1 is frequently reported to be dysregulated in the cancer including neuroendocrine tumors, lung cancer and melanoma. However, the role of MEN1 in case of Breast cancer has not been well understood. Lifestyle, environmental conditions and ethnicity play crucial role in epigenetic modulation of gene expression. In the current scenario with high increase in number of cancer cases, there is need to develop more personalized course of treatment rather than generalized approach. Thus, discovery of new biomarkers will aid in designing better diagnostic and therapeutic strategy to treat cancer. Aims and Objective: In this study we aim to find MEN1 mRNA expression and its connection with the MEN1 promoter methylation in case of Indian breast cancer patients and to correlate our findings with the pathological parameters of the patients to understand the role of MEN1 gene in breast cancer progression. Material and methods: The ethical approval for the study was procured from Institutional ethical committee of AIIMS,New Delhi and Jamia Millia Islamia, New Delhi. The study comprises of 44 pair of Breast tissue samples including cancer and adjacent normal tissue collected at department of surgical oncology AIIMS, New Delhi and were stored in RNA later and PBS at -80˚C for RNA and DNA extraction respectively. DNA was extracted through PCI (Phenol-Chloroform-Isoamyl alcohol) method and was further confirmed by Agarose gel electrophoresis. TRIZOL method was used to extract RNA from tissue samples and was quantified using nanodrop. Moreover, cDNA synthesis was carried out using Verso cDNA synthesis kit. To investigate the MEN1 mRNA expression, RT- PCR was carried out with LightCycler96 SYBR Green I Master (Roche) using specific primers. To analyze the promoter methylation status of MEN1 promoter region, CT- conversion was done and MS-PCR was performed using Methylation and Unmethylation specific primers. Statistical analysis was done using Graph Pad and SPSS to correlate our findings with clinical parameters. Results: In our study we found 27 out of 44 patients to exhibit up-regulation of MEN1 mRNA in the breast cancer tissue as compared with the normal tisuue. MEN1 promoter unmethylation was detected in 25 out of 44 cases, which revealed its significant correlation with the upregulated MEN1 gene. Further on correlation of our data with the clinical parameters, we found its significant association with the estrogen receptor status and age of menopause of the patients. Conclusion: Our results of the MEN1 mRNA expression study and its association with MEN1 promoter methylation in case of breast cancer suggests the tumorigenic role of MEN1 in breast cancer. Our findings thereby indicate the possible role of MEN1 gene in progression of the disease. Further study on large sample size will aid in better understanding the role of MEN1 gene in Breast carcinoma. Citation Format: Sheersh Massey, Aasif Khan, Asifa Khan, SVS Deo, Syed Akhtar Husain. Promoter hypomethylation associated upregulation of MEN1 mRNA expression and its correlation with pathological parameters in Indian breast cancer patients [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-18-02.