miR-7155-3p在乳腺癌患者血清中的表达及对乳腺癌细胞增殖和侵袭的影响

Expression of miR-7155-3p in the serum of breast cancer patients and its effect on the proliferation and invasion of breast cancer cells

目的分析微小RNA(miRNA,miR)-7155-3p在乳腺癌患者血清中的表达,探讨其对乳腺癌细胞增殖和侵袭的影响及作用机制。方法采用实时定量聚合酶链反应(RT-PCR)检测miR-7155-3p在乳腺癌患者血清和乳腺癌细胞系中的表达。以miR-7155-3p表达最少的细胞系为对象,分别转染人工合成miR-NC(阴性对照序列)和miR-7155-3p模拟物,命名为miR-NC组和miR-7155-3p组。采用生物信息学软件预测及双荧光素酶报告实验验证miR-7155-3p的下游靶基因。RT-PCR和Western blot检测下游靶基因的表达。采用噻唑蓝(MTT)实验和Transwell实验检测乳腺癌细胞增殖和侵袭能力。结果与正常人血清相比,miR-7155-3p在乳腺癌患者血清中的表达明显降低(P<0.01)。与正常乳腺上皮细胞相比,miR-7155-3p在乳腺癌细胞系的表达明显降低(P<0.01),在SKBR3细胞中表达最少(P<0.01)。生物信息学软件显示miR-7155-3p的下游靶基因是Tribbles同源蛋白1(TRIB1)。双荧光素酶报告实验证明miR-7155-3p可互补结合TRIB1基因(P<0.01)。与miR-NC组相比,miR-7155-3p组细胞中TRIB1基因的表达明显降低(P<0.01),细胞增殖能力明显降低(P<0.05),细胞侵袭能力明显降低(P<0.01)。结论miR-7155-3p在乳腺癌患者血清中表达降低,miR-7155-3p可通过转录后下调TRIB1基因的表达抑制乳腺癌细胞SKBR3的增殖和侵袭能力。

Objective To analyze the expression of microRNA(miRNA,miR)-7155-3p in the serum of breast cancer patients,and explore its effect on breast cancer cell proliferation and invasion and its mechanism of action.Methods Real-time PCR(RT-PCR)was used to detect the expression of miR-7155-3p in breast cancer patients’serum and breast cancer cell lines.The cell lines with the least expression of miR-7155-3p were taken as targets,and the synthetic miR-NC(Negative control sequence)and miR-7155-3p mimics were transfected respectively,and named as miR-NC group and miR-7155-3p group.Bioinformatics software and dual luciferase report experiment were used to predict and verify the downstream target genes of miR-7155-3p.RT-PCR and Western blot were used to detect the expression of downstream target genes.The thiazole blue(MTT)test and Transwell test were used to detect the proliferation and invasion ability of breast cancer cells.Results Compared with normal human serum,the expression of miR-7155-3p in breast cancer patients was significantly reduced(P<0.01).Compared with normal breast epithelial cells,the expression of miR-7155-3p in breast cancer cell lines was significantly reduced(P<0.01),and the expression was minimal in SKBR3 cells(P<0.01).Bioinformatics software and dual luciferase report experiment identified that the downstream target gene of miR-7155-3p was Tribbles homologous protein 1(TRIB1)(P<0.01).Compared with the miR-NC group,the expression of TRIB1 gene in cells of miR-7155-3p group was significantly reduced(P<0.01).Moreover,the cell proliferation ability and invasive ability were significantly reduced(P<0.05).Conclusion The expression of miR-7155-3p is reduced in the serum of breast cancer patients.miR-7155-3p can inhibit the proliferation and invasion of breast cancer cells SKBR3 by down-regulating the expression of TRIB1 gene.