miR-135b基因对HCT116结肠癌细胞增殖及侵袭的影响

Effect of miR-135b gene on cell proliferation and invasion of HCT116 colorectal cancer cells

目的探讨miR-135b基因对HCT116结肠癌细胞增殖及侵袭的影响。方法用Realtime PCR法检测HCT116、Caco-2、HT-29、SW480结肠癌细胞及HCoEpiC人正常结肠上皮细胞中miR-135b表达。细胞分为正常组、miR-135b NC组及miR-135b抑制组,噻唑蓝(MTT)实验检测细胞活力,Realtime PCR法检测miR-135b表达,Transwell法检测细胞侵袭能力,western blot法检测B淋巴细胞瘤-2基因(Bcl-2)、增殖细胞核抗原(PCNA)、基质金属蛋白酶-2(MMP-2)及MMP-9蛋白表达量。结果 miR-135b基因在结肠癌细胞HCT116(2.49±0.23)、HT-29(1.89±0.19)、SW480(1.76±0.18)和Caco-2(1.59±0.13)中的表达量,高于HCoEpiC正常结肠上皮细胞(1.00±0.12);与正常组及miR-135b NC组比较,miR-135b抑制组中miR-135b表达量下降,细胞活力降低,细胞侵袭数减少,Bcl-2、PCNA、MMP-2和MMP-9蛋白表达量降低。结论 miR-135b基因表达量下调,可抑制HCT116结肠癌细胞的增殖及侵袭。

Objective To explore the effect of miR-135 b gene on cell proliferation and invasion of HCT116 colorectal cancer cells. Methods The expression of miR-135 b gene in HCT116, Caco-2, HT-29, SW480 colorectal cancer cells and HCoEpiC human normal colorectal epithelial cells was detected by Realtime PCR method. The cells were divided into normal group, miR-135 b NC group and miR-135 b inhibitor group. The cell viability was detected by MTT method. The expression of miR-135 b was detected by Realtime PCR method. The cell invasion ability was detected by Transwell method. The expression of B-cell lymphoma-2(Bcl-2), proliferating cell nuclear antigen(PCNA), matrix metalloproenases-2(MMP-2) and MMP-9 protein were detected by western blot method. Results The expression quantities of miR-135 b gene in HCT116(2.49±0.23), HT-29(1.89±0.19), SW480(1.76±0.18) and Caco-2(1.59±0.13) cells were higher than that in normal colorectal epithelial cells(1.00±0.12). Compared with normal group and miR-135 b NC group, the expression quantity of miR-135 b was decreased, cell viability and cell invasion number was reduced, the expression of Bcl-2, PCNA, MMP-2 and MMP-9 protein were decreased. Conclusion The decreasing expression quantity of miR-135 b gene can inhibit HCT116 cells proliferation and invasion.