Vishwajeet Rohil
Vallabhbhai Patel Chest Institute, India
Title: Anticancer activity of polyphenolic acetates mediated by calreticulin transacetylase in lung cancer
Biography
Biography: Vishwajeet Rohil
Abstract
Background: Cancer is a group of diseases involving abnormal cell growth, occasionally having metastasis. This underlying genetic disease is initiated either by mutation or epigenetics. We are targeting lung carcinoma, most common cause of cancer-related death in men and women. Our new drug discovery is targeted on Calreticulin Transacetylase (CRTAase). We intend to treat lung cancer in-vivo and in-vitro by inducing hyperacetylation and upregulating the expression of genes important in tumor suppression. The polyphenolic acetates in combination with HDAC inhibitors are known to promote hyperacetylation, leading to apoptosis in lung cancer cells.
Aim: To determine the anticancer activity of 7, 8-Diacetoxy-4-Methyl Coumarin (DAMC), Polyphenolic acetate targeting acetylated histone interaction.
Methodology: DAMC showed anticancer activity both in-vitro and in-vivo. The hyperacetylation activity of DAMC on CRTAase induced epigenetic modulations were observed in A549 cells, as well as mice with Ehrlich ascites tumor (EAT) cells. The in-vitro and in-vivo data was validated by the apoptosis. Additionally specific target based anticancer property of DAMC was evaluated using microarray and RTPCR prior and after demethylation.
Results: In A549 cells, highest transfection efficiency was obtained after 72 hrs. Significant increase (p<0.01) in expression of H3 (2.67±0.02) and H4 2.755±0.016) was observed in DAMC treated CRTAase gene transfected A549 cells as compared to non-transfected A549 cells treated with DAMC (2.14±0.023) and (2.161±0.011) respectively. High apoptotic index was observed in The EAT cells in-vivo as well as in A549 cells in-vitro. RNA having RIN (RNA Integrity) values between 8.5 and 9.8 on electropherogram were subjected to microarray and RTPCR. A549 cells treated with DAMC and Valproic acid (VA) were suggestive of synergistic upregulation of tumor suppressor genes viz. ING4, TCF21, MFSD2A, FHIT and metalloproteinase inhibitor 3 i.e.TIMP3 and downregulating the oncogene Skp2.
Conclusion: The in-vivo as well as in-vitro findings suggest that DAMC and VA can potentiate the apoptotic pathway via CRTAase and thus can be a very promising anticancer drug candidate. In further ongoing studies we are screening more drugs targeting similar/more molecular targets and extending correlation with clinical applications.