miR-101对结直肠癌细胞增殖、凋亡及KIF14蛋白表达的影响

Effect of miR-101 on proliferation,apoptosis and KIF14 expression of colorectal cancer cells

目的:研究miR-101对结直肠癌细胞增殖、凋亡及驱动蛋白家族蛋白14(KIF14)表达的影响。方法:利用靶基因预测库预测miR-101与KIF14的靶向关系,并采用双荧光素酶报告实验鉴定。采用实时荧光定量PCR和Western blot法测定结直肠癌组织和细胞中miR-101和KIF14蛋白的表达。在结直肠癌HT29细胞中转染miR-101模拟物,MTT法测定细胞增殖情况,Annexin V-FITC/PI双染法检测细胞凋亡率,Western blot法测定KIF14蛋白的表达。结果:双荧光素酶报告实验结果显示,miR-101与KIF14存在靶向关系。结直肠癌组织和细胞中miR-101表达下调,KIF14表达上调(P <0. 05)。miR-101模拟物可以明显抑制HT29细胞的增殖能力并促进其凋亡,同时下调KIF14蛋白表达水平(P <0. 05)。结论:miR-101可靶向作用于KIF14而影响结直肠癌细胞的增殖和凋亡。

Aim: To study the effect of miR-101 on proliferation, apoptosis and KIF14 expression of colorectal cancer cells. Methods: Target gene prediction library was used to predict the targeting relationship between miR-101 and KIF14, and double luciferase reporter assay was used to identify the relationship. The expressions of miR-101 and KIF14 in colorectal cancer tissue and colorectal cancer cells were detected by qRT-PCR and Western blot. miR-101 mimics was transfected into HT29 cells, and then cell proliferation was measured by MTT, apoptosis was detected by Annexin V-FITC/PI staining, Western blot was used to detect the KIF14 protein expression. Results: The double luciferase reporter assay showed that there was a target relationship between miR-101 and KIF14. miR-101 was downregulated and KIF14 was upregulated in colorectal cancer tissue and colorectal cancer cells( P < 0. 05) . miR-101 mimics significantly inhibited the proliferation and KIF14 protein expression, and promote apoptosis in HT29 cells( P < 0. 05) . Conclusion: miR-101 could inhibit proliferation and induce apoptosis of colorectal cancer cells by targeting KIF14.