miR-92b-3p对结直肠癌细胞增殖的影响及其可能机制的初步探讨

MicroRNA-92b-3p regulates colorectal cancer cell proliferation and its possible mechanism

目的探讨miR-92b-3p对结直肠癌细胞增殖的影响及其可能的作用机制。方法检测miR-92b-3p在结直肠癌组织、癌旁正常组织、人结直肠癌HCT116细胞株及人正常结直肠黏膜细胞(FHC)的表达。在HCT116细胞株中转染miR-92b-3p inhibitor,采用CCK-8检测其对细胞增殖的影响。通过生物信息学方法分析miR-92b-3p的下游靶基因并用双荧光素酶报告基因法验证。采用CCK-8检测靶基因过表达对HCT116细胞增殖的影响。设置空白对照组(blank)、阴性对照组(NC)、miR-92b-3p inhibitor组及miR-92b-3p inhibitor+靶基因siRNA组,观察对HCT116细胞增殖的影响。结果 miR-92b-3p在结直肠癌组织中的表达水平高于癌旁正常组织,在HCT116细胞株中表达水平高于FHC (均P <0.05)。抑制miR-92b-3p表达后HCT116细胞OD值下降(P <0.05)。FBXW7为miR-92b-3p的靶基因,FBXW7过表达可抑制HCT116细胞增殖(P <0.05)。在培养48 h、72 h时,与blank、NC比较,miR-92b-3p inhibitor组HCT116细胞OD值下降,miR-92b-3p inhibitor+FBXW7 siRNA组HCT116细胞OD值高于miR-92b-3p inhibitor组(均P <0.05)。结论 miR-92b-3p在结直肠癌的发生发展过程中起到促癌基因的作用,其可能通过调控FBXW7影响结直肠癌细胞的增殖。

Objectives To investigate the effect of miRNA-92b-3p on colorectal cancer cell proliferation and its possible mechanism. Methods Expression of miR-92b-3p in colorectal cancer tissue, adjacent normal tissue, human HCT116 colorectal cancer cell line, and normal fetal colonic mucosa(FHC) cell were measured. MiR-92b-3p inhibitor was transfected into HCT116 cell line and cell proliferation was measured by Cell Counting Kit-8(CCK-8). Bioinformatics was used to analyze target gene of miR-92b-3p and the target gene were verified by dual luciferase reporter gene assay. CCK-8 was then used to evaluate the effect of target gene overexpression on HCT116 cell line proliferation. Four groups were established to investigate the effect of miR-92b-3p on HCT116 cell proliferation: blank, negative control(NC), miR-92b-3p inhibitors, miR-92b-3p inhibitors plus siRNA of target gene. Results MiR-92b-3p was significantly higher in colorectal cancer tissue than in adjacent normal tissue, in HCT116 cell line than in FHC cell(P < 0.05). The OD(optical density) value decreased significantly after inhibiting miR-92b-3p(P < 0.05). FBXW7 was the target gene of miR-92b-3p. Overexpression of FBXW7 could inhibit HCT116 cell proliferation(P < 0.05). Compared with the blank and NC groups, the OD value of HCT116 cell decreased in miR-92b-3p inhibitor group at 48 h and 72 h(P < 0.05). The OD value was significantly higher in the miR-92b-3p inhibitor+FBXW7 siRNA group than in the miR-92b-3p inhibitor group(P < 0.05). Conclusion MiR-92b-3p promoted carcinogenesis in the development and progression of colorectal cancer. It could influence cancer cell proliferation by regulating FBXW7.