lncRNA SNHG6对三阴性乳腺癌细胞增殖、迁移及凋亡的影响研究及其ceRNA网络初探

Effects of lncRNA SNHG6 on proliferation,migration and apoptosis in triple-negative breast cancer cells by regulating miR-519d-3p/CCND1

目的 分析长链非编码RNA(lncRNA)SNHG6靶向调控miR-519d-3p/CCND1对三阴性乳腺癌(TNBC细胞增殖、迁移及凋亡的影响。方法 采用siRNA技术沉默TNBC细胞系MDA-MB-231中lncRNA SNHG6的表达,实验分为三组:正常人乳腺上皮细胞系MCF-10A(对照组)、MDA-MB-231和si-SNHG6(si组),采用实时荧光定量PCR(RT-qPCR)检测lncRNA SNHG6和miR-519d-3p表达,MTT法检测细胞增殖率,Transwell实验检测迁移能力,流式细胞术检测细胞凋亡率,Western blot实验检测CCND1蛋白表达。采用双荧光素酶报告基因检测SNHG6与miR-519d-3p及miR-519d-3p与CCND1的靶向关系。三组间计量资料采用单因素ANOVA分析和LSD-t法检验,P<0.05为差异有统计学意义。结果 连续培养48 h发现,与对照组比较,MDA-MB-231组lncRNA SNHG6与miR-519d-3p mRNA相对表达量、细胞增殖率、迁移率和CCND1蛋白水平均显著升高,细胞凋亡率显著下降(P<0.05);与MDA-MB-231组比较,si组lncRNASNHG6与miR-519d-3pmRNA相对表达量显著降低、细胞增殖率、迁移率和CCND1蛋白水平均显著下降(P<0.05),凋亡率上升;双荧光素酶报告基因检测显示,SNHG6能够靶向上调miR-519d-3p表达,miR-519d-3p能够靶向上调CCND1表达。结论 lncRNASNHG6能够通过靶向调控miR-519d-3p/CCND1功能进而影响TNBC细胞的增殖、迁移及凋亡能力,可能是TNBC发病的重要分子机制,也有望成为分子干预的靶点。

Objective To analyze the effects of long-chain non-coding RNA(lncRNA) SNHG6 targeting miR-519d-3p/CCND1 on the proliferation,migration and apoptosis of triple-negative breast cancer(TNBC) cells.Methods siRNA technology was used to silence the expression of lncRNA SNHG6 in the TNBC cell line MDA-MB-231.The experiment was divided into three groups:Normal human breast epithelial cell line MCF-10A(control group),MDA-MB-231 and si-SNHG6(si group),real-time fluorescent quantitative PCR(RT-qPCR) was used to detect the expression of lncRNA SNHG6 and miR-519d-3p,MTT method was used to detect cell proliferation rate,Transwell test was used to detect migration ability,flow cytometry was used to detect cell apoptosis rate,Western blot the experiment detects the expression of CCND1 protein.The dual luciferase reporter gene was used to detect the targeting relationship between SNHG6 and miR-519d-3p and miR-519d-3p and CCND1.The measurement data among the three groups were analyzed by one-way ANOVA and LSD-t test and P<0.05indicated that the difference was statistically significant.Results After 48 hours of continuous culture,compared with the control group,the relative expression of lncRNA SNHG6 and miR-519d-3p mRNA,cell proliferation rate,migration rate and CCND1 protein level in the MDA-MB-231 group were significantly increased and the apoptosis rate was significantly increased(P<0.05).Compared with the MDA-MB-231 group,the relative expression of lncRNA SNHG6 and miR-519d-3p mRNA in the si group was significantly decreased and the cell proliferation rate,migration rate and CCND1 protein level were significantly decreased,and the death rate increased(P<0.05).Dual luciferase reporter gene detection showed that SNHG6 can target up-regulating the expression of miR-519d-3p and miR-519d-3p can target up-regulating the expression of CCND1.Conclusion lncRNA SNHG6 can affect the proliferation,migration and apoptosis of TNBC cells through targeted regulation of miR-519d-3p/CCND1 function.It may be an important molecular mechanism for the pathogenesis of TNBC and it is also expected to become a target of molecular intervention.