Molecular Mechanism Of MiR-1291 In Pancreatic Cancer Cells And Its Effect On Apoptosis

Objective:To elucidate the possible molecular pathway of miR-1291 in pancreatic cancer cells and its effect on apoptosis,and to study the mechanism of miR-1291 occurrence and development in pancreatic cancer,in order to provide a new direction for early diagnosis and gene targeted therapy of pancreatic cancer.Methods:The expression of miR-1291 in pancreatic duct epithelial cells(HPDE6-C7)and pancreatic cancer cells(PANC-1)was examined by qRT-PCR in this study.miR-1291mimics(miR-1291 mimics),miR-1291 inhibitor(miR-1291 inhibitor)and control groups(miR-1291 mimics NC,miR-1291 inhibitor NC)were transfected into pancreatic cancer cell lines using LIPO 3000 in vitro,the control group(no transfection gene was added,the others were the same as the experimental group).The miR-1291 target gene and its expression protein were predicted by bio-informatics analysis,and the miR-1291 analogue,miR-1291 inhibitor and total cell protein after miR-1291 transfection were extracted by qRT-PCR,western blot was used to detect the expression of related target proteins,and the effect of miR-1291 analogue,miR-1291 inhibitor and control group on apoptosis of pancreatic cancer cells was detected by Flow cytometry.Results:(1)qRT-PCR results showed that the expression level of pancreatic cancer cells(PANC-1)miR-1291 was lower than that of pancreatic ductal epithelial cells (HPDE6-C7),the expression level of pancreatic cancer cells(miR-1291 mimics)was significantly increased in transfection miR-1291 mimic group(blank group),the expression level of pancreatic cancer cells(miR-1291 inhibitor)was significantly decreased in transfection inhibitor group(control group),and miR-1291mimics NC、miR-1291 inhibitor NC).There was no significant difference in miR-1291 expression in pancreatic cancer cells.(2)Using bio-informatics analysis technology,it is predicted that miR-1291 may target genes such as ROCK-1、MUC1 to play its role in tumor suppressor in pancreatic cancer.To further clarify miR-1291 regulation of predictive proteins in pancreatic cancer cell lines(PANC-1),Protein imprinting was used to detect the expression of ROCK-1、MUC1 after transfection miR-1291 analogue group(miR-1291 mimics),miR-1291 inhibitor group(miR-1291 Inhibitor)and control group(miR-1291 mimics NC 、miR-1291 inhibitor NC).The results showed that after transfection miR-1291 mimics NC and miR-1291 inhibitor NC,and there was no significant difference in the expression of ROCK-1 and MUC1 protein,but the expression of ROCK-1 and MUC1 protein was significantly reduced in the transfection miR-1291 mimics group,and the expression of ROCK-1 and MUC1 protein was significantly up-regulated in the transfection group.(3)Flow cytometry detection of apoptosis: compared with the blank control group,transfection miR-1291 mimic group(miR-1291mimics)significantly promoted pancreatic cancer cell apoptosis(early apoptosis late apoptosis),while miR-1291inhibitor(miR-1291Inhibitor)inhibited pancreatic cancer cell apoptosis(early apoptosis late apoptosis).Conclusion:(1)miR-1291 in pancreatic duct epithelial cells and pancreatic cancer cells there is differential expression,and in pancreatic duct epithelial cells expression level is much higher than in pancreatic cancer cells;(2)Through up-regulating or down-regulating the expression level of miR-1291 in pancreatic cancer cells,thus affecting the expression level of ROCK-1、MUC1,indirectly from the protein level verification ROCK-1 and MUC1 as the downstream target gene regulatory products,that is,in pancreatic cancer cells,The biological function of pancreatic cancer is affected by miR-1291/ROCK-1 and miR-1291/MUC1 pathways;(3)Up-regulation of miR-1291 expression in pancreatic cancer cells can significantly promote apoptosis of pancreatic cancer cells,while down-regulation of miR-1291 expression in pancreatic cancer cells can significantly inhibit apoptosis of pancreatic cancer cells.