miR⁃429靶向调控SIAH1基因对结直肠癌细胞LOVO增殖的影响

Effects of miR⁃429 on proliferation of colorectal cancer cell line LOVO via targeting SAIH1 gene

目的探讨miR⁃429靶向SIAH1基因对结直肠癌细胞系LOVO细胞增殖的影响。方法采用qRT⁃PCR检测正常结肠黏膜上皮细胞系NCM460和结直肠癌细胞系SW480、LOVO、HT⁃29中miR⁃429的表达。将miR⁃429 mimic和miR⁃429阴性对照序列(NC)转染至LOVO细胞,同时设置Control组。预测miR⁃429的靶基因并用双荧光素酶报告基因实验验证靶向关系。采用Western blot检测SIAH1蛋白的表达水平,MTT法检测细胞增殖情况,流式细胞术检测细胞周期分布情况。结果qRT⁃PCR检测结果显示,相对于NCM460细胞,miR⁃429在各结直肠癌细胞系中的表达均降低(均P<0.001)。双荧光素酶报告基因实验证实了SIAH1与miR⁃429的靶向关系。与NC组相比,过表达miR⁃429能抑制LOVO细胞中SIAH1蛋白的表达(P<0.001),细胞被阻滞在G0G1期,细胞增殖能力降低(P<0.001)。结论miR⁃429通过靶向SIAH1基因表达抑制LOVO细胞增殖。

Objective To investigate the effects of miR⁃429 targeting SIAH1 gene on the proliferation of colorectal cancer cell line LOVO.Methods The expression of miR⁃429 in the normal colonic epithelial cell line NCM460 and colorectal cancer cell lines SW480,LOVO and HT⁃29 were detected by qRT⁃PCR.miR⁃429 mimic and miR⁃429 negative control sequence(NC)were transfected into LOVO cells,and the control group was set.The target gene of miR⁃429 was predicted,and the targeting relationship was verified by the dual luciferase reporter assay.The expression level of SIAH1 protein was detected by Western blot,cell proliferation was detected by MTT assay,and the cell cycle distribution was detected by flow cytometry.Results qRT⁃PCR showed that,compared with NCM460 cells,the expression of miR⁃429 was decreased in all the colorectal cancer cell lines(all P<0.001).The dual luciferase reporter assay confirmed that SIAH1 had a targeting relationship with miR⁃429.Compared with the NC group,overexpression of miR⁃429 inhibited the expre⁃ssion of SIAH1 protein in LOVO cells(P<0.001),the cell cycle were blocked in G0G1 phase,and cell proliferative was reduced(P<0.001).Conclusions miR⁃429 can inhibit the proliferation of LOVO cells by targeting SIAH1 gene expression.