MicroRNA-375 Overexpression Of Lentivirus Vector Regulates EMT To Inhibit The Invasion And Migration Of Colorectal Cancer Cells

Objective: To construct microRNA-375 overexpressing colorectal cancer cell HCT116 by lentivirus vector,and study the role of microRNA-375 in the invasion and migration of colorectal cancer cells,and the relationship between microRNA-375 and EMT during the invasion and migration of colorectal cancer cells.Methods: The colorectal cancer tissues and adjacent normal tissues confirmed by pathology were divided into adjacent normal tissues,early tumor tissues and advanced tumor tissues according to pathological grading.The expression of micro RNA-375 in 3 cells was detected by real time reverse transcriptase PCR.MicroRNA-375 overexpression of HCT116 cells was constructed and cultured by lentivirus vector,and an empty vector control group and a blank control group were set up at the same time.Real time reverse transcriptase PCR was used to detect the expression of microRNA-375 in 3 groups of cells.The Trans-well compartment model reflected the invasiveness of the 3 groups of cells.Scratch test reflected the migration of 3 groups of cells.The relative content of protein in 3 groups of EMT cells was detected by Western blot Western blot.The results were statistically processed,and P<0.05 indicated that difference was statistically significant.Results: The relative expression of microRNA-375 in normal tissue(0.990 ± 0.120)adjacent to cancer of colorectal cancer was higher than that in early tumor tissue(0.630 ± 0.095)and late tumor tissue(0.400 ± 0.125),the difference was statistically significant(P <0.05),and microRNA-375 showed high expression;the microRNA-375 of colorectal cancer tissues showed low expression,and the relative expression of microRNA-375 in early tumor tissues was higher than that of late tumor tissue(P<0.05).Lentivirus vector was transfected successfully,and HCT116 cells of microRNA-375 overexpressing experimental group and empty vector control group were constructed and cultured.The relative expression of microRNA-375 in the over expression group(143.300 ± 7.889)was significantly higher than that in the empty carrier group(1.010 ± 0.191)and the blank group(1.000 ± 0.000),the difference was statistically significant(P <0.05);there was no significant difference in the relative expression of microRNA-375 between emptyand blank groups(P>0.05).The number of overexpressed group(277.000 ± 9.192)in the Trans-well compartment model was smaller than that in the empty carrier group(572.600 + 14.100)and the blank group(583.400 ± 12.661),the difference was statistically significant(P <0.05);there was no significant difference in the number of penetrating cells between empty group and blank group(P>0.05).The wound healing rate of the overexpressed group(0.440 ± 0.016)in the scratch test was lower than that of the empty carrier group(0.660 ± 0.029)and the blank group(0.684 ± 0.021),and the difference was statistically significant(P <0.05);there was no statistically significant difference in the healing rate of the scratches of the cells in the empty and blank groups(P>0.05).The ratio of E-cadherin in the over expression group(1.202 ± 0.035)in Western blot was higher than that in the empty carrier group(0.970 ± 0.016)and the blank group(0.942 + 0.029),the difference was statistically significant(P <0.05);there was no significant difference in the E-cadherin gray value ratio between the empty and the blank groups(P>0.05).The ratio of Vimentin in the over expression group(0.574 ± 0.040)was lower than that of the empty carrier group(1.076 ± 0.036)and the blank group(1.060 ± 0.035),the difference was statistically significant(P <0.05);there was no significant difference in the ratio of Vimentin gray value between the empty and the blank groups(P>0.05).The EMT process of the overexpressing group was inhibited compared with the empty carrier group and the blank group(P <0.05);there was no difference in the EMT process between the empty carrier group and the blank group(P>0.05).Conclusion: MicroRNA-375 showed low expression in colorectal cancer cells.The higher the malignancy of colorectal cancer cells,the lower the expression of microRNA-375.As a tumor suppressor factor,microRNA-375 can inhibit the invasion and migration of colorectal cancer cells in human body by inhibiting EMT.