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目的探讨Hsa-miR-193b对人子宫内膜腺癌细胞系HEC-1B侵袭的影响及其相关机制。方法将Hsa-miR-193b模拟物(mimics)、Hsa-miR-193b抑制剂(inhibitor)转染HEC-1B细胞,Real-time PCR证实转染是否成功;Transwell小室法检测转染Hsa-miR-193b mimics后HEC-1B细胞的侵袭能力;通过miRGen、Targetscan、Pictar数据库筛查Hsa-miR-193b指向与侵袭相关的靶基因;Real-time PCR检测转染Hsa-miR-193b mimics转染组和对照组mRNA水平;Western blot检测各组GRB7蛋白表达的水平。结果①Hsa-miR-193b mimics、Hsa-miR-193b inhibitor转染HEC-1B细胞成功,两转染组Hsa-miR-193b表达水平与未转染组比较差异有统计学意义(P<0.05)。②转染Hsa-miR-193b mimics组与对照组比较,肿瘤细胞的侵袭能力明显降低(P<0.05)。③转染后,靶基因GRB7蛋白Hsa-miR-193b mimics组表达下降(P<0.05)、Hsa-miR-193b inhibitor组表达增加(P<0.05),且各组GRB7 mRNA水平无显著变化。结论在HEC-1B中转染Hsa-miR-193b mimics能下调GRB7蛋白表达,并抑制HEC-1B细胞的侵袭。
Objective To investigate the effect of Hsa-miR-193b on invasion of human endometrial adenocarcinoma cell line HEC-1B and its related mechanism. Methods Hsa-miR-193b mimics and Hsa-miR-193b inhibitor were transfected into HEC-1B cells. Real-time PCR was used to confirm whether transfection was successful or not. 193b mimics; Hsa-miR-193b was screened by miRGen, Targetscan, Pictar database for target genes related to invasion; Real-time PCR was used to detect the expression of Hsa-miR-193b mimics transfection group and The mRNA level of GRB7 in each group was detected by Western blot. Results ① Hsa-miR-193b mimics and Hsa-miR-193b inhibitor were successfully transfected into HEC-1B cells. The expression levels of Hsa-miR-193b in two transfected groups were significantly different from those in untransfected group (P <0.05). ② Compared with the control group, the invasion ability of Hsa-miR-193b mimics group was significantly decreased (P <0.05). The expression of GRB7 protein in Hsa-miR-193b mimics group decreased (P <0.05) and the expression of Hsa-miR-193b inhibitor group increased (P <0.05) after transfection. There was no significant change in GRB7 mRNA level in each group. Conclusion Hsa-miR-193b mimics can down-regulate the expression of GRB7 protein and inhibit the invasion of HEC-1B cells in HEC-1B.