Molecular Mechanism Of Colorectal Cancer Cells Apoptosis Induced By Metformin Combined With Radiotherapy

Objective:To observe ADORA1 genes in metformin combined radiotherapy induced colorectal cancer SW480 and HCT116 cells proliferation and apoptosis,and to explore its possible mechanisms. Methods:1. Using microarray analysis SW480 and HCT116 human colorectal cancer cells in the control group, one with metformin, a single X-ray group, metformin differentially expressed genes in the X-ray group, screening out the significant differences and apoptosis related genes, providing new clues to prepare and study the mechanism for our follow-up studies.2. Apply to colony formation assay and flow cytometry to detect the metformin alone, single-use X-rays, X-rays metformin, metformin DPCPX(ADORA1 inhibitors) and other descendants of different interventions survival and apoptosis in colorectal cancer cells.3. Using the Quantitative PCR validation of microarray analysis after treatment in different ways, m RNA expression in cells ADORA1, Caspase3, Caspase9, Bcl-2’s.4. Apply to Western Blotting to detect the expression of different ways related proteins after treatment of the cells. Results:1.Colony formation assay results showed that: metformin alone(1mmol / l) inhibited cell viability, whereas metformin(1mmol / l) and the combined group DPCPX inhibition of cell viability was significantly lower than the group alone, metformin when cloning DPCPX group on cell formation inhibition was significantly lower than metformin alone, and the difference was statistically significant(P <0.05).2.Flow cytometry showed that: cells with metformin alone or metformin in combination with X-ray group after joining DPCPX, induced apoptosis was significantly decreased with significant difference(P <0.05).3.Quantitative PCR results showed that: verify the results of gene chip in metformin(1mmol / l) and X-ray(8Gy) combined group ADORA1, Caspase3, m RNA expression were higher than Caspase9 alone group, and low expression of Bcl-2 m RNA in the monotherapy group, and there is significant difference(P <0.05).4.Western blotting results showed that: with metformin alone(1 mmol / l), a single X-ray group(8Gy) compared with the combined group of p-AMPK, ADORA1, Caspase3, Caspase9 expression was significantly increased, while the p-m TOR and Bcl- 2 expression was significantly decreased. Conclusion:1.Metformin inhibits SW480 and HCT116 human colorectal cancer cell proliferation and viability, but metformin DPCPX group inhibition of cell viability was significantly lower than metformin alone.2.Metformin combined with X-ray promote SW480 and HCT116 human colorectal cancer cell apoptosis was significantly stronger than the effect of apoptosis induced by both when used alone, but with metformin alone or metformin in combination with X-ray group added DPCPX after induction of cell apoptosis was significantly decreased.3.From the study, we found that metformin combine with ADORA1 induce apoptosis in human colorectal cancer cells.