LncRNA SLC16A1-AS1靶向调控miR-182对乳腺癌BT549细胞增殖能力的影响  

LncRNA SLC16A1-AS1 targeting regulation of miR-182 inhibits the proliferation of breast canceRBT549 cells

作　　者：蒋冰[1] 马琳[1] 刘骞[1] JIANG Bing;MA Lin;LIU Qian(Department of Pathology,Liaoning CanceRHospital&Institude,Shenyang 110042,China)

机构地区：[1]辽宁省肿瘤医院病理科,沈阳110042

出　　处：《临床与实验病理学杂志》2023年第8期909-914,共6页Chinese Journal of Clinical and Experimental Pathology

摘　　要：目的探讨LncRNA SLC16A1-AS1对miR-182的靶向调控作用及对乳腺癌BT549细胞增殖能力的影响。方法使用GEPIA2工具分析TCGA数据库中LncRNA SLC16A1-AS1在三阴型乳腺癌(triple negative breast cancer,TNBC)中的差异性表达,应用Kaplan-MeieRPlotter数据库进行生存分析。利用RegRNA2和miRanda数据库预测LncRNA SLC16A1-AS1的潜在靶标miRNAs。用qRT-PCR检测LncRNA SLC16A1-AS1在TNBC组织及细胞系(BT549、BT20、MDA-MB-231及MDA-MB-468)中的表达。利用双荧光素酶活性测定及RNA免疫沉淀实验验证LncRNA SLC16A1-AS1与miR-182的结合力。用CCK-8及集落形成实验检测不同处理组BT549细胞增殖能力的变化。结果生物信息学分析结果显示,TNBC组织中LncRNA SLC16A1-AS1的表达水平显著低于正常组织(P<0.001),与预后良好相关(P<0.001),LncRNA SLC16A1-AS1与miR-182之间存在结合位点。LncRNA SLC16A1-AS1在63例TNBC组织中的表达水平显著低于瘤旁组织(7.94%vs 63.49%,P<0.001),在TNBC细胞系中的表达水平亦显著低于正常乳腺上皮细胞系(P<0.01)。在BT549细胞中过表达miR-182可显著降低LncRNA SLC16A1-AS1-WT载体的荧光酶素活性,RNA免疫沉淀实验显示,AGO2同时富集LncRNA SLC16A1-AS1及miR-182(P<0.01)。与对照组相比,过表达LncRNA SLC16A1-AS1显著降低BT549细胞的增殖活性并减少集落形成数量,共表达LncRNA SLC16A1-AS1与miR-182可恢复BT549细胞的增殖能力(P<0.01)。结论LncRNA SLC16A1-AS1在TNBC中低表达,且通过靶向调控miR-182在体外抑制TNBC细胞的增殖能力,LncRNA SLC16A1-AS1/miR-182轴有望成为TNBC治疗的潜在靶点。Purpose To investigate the targeting regulation of LncRNA SLC16A1-AS1 on miR-182 and its effect on the proliferation of breast canceRBT549 cells.Methods GEPIA2 tool was used to analyze the differential expression of LncRNA SLC16A1-AS1 in triple negative breast canceR(TNBC)and Kaplan-MeieRPlotteRdatabase was used foRsurvival analysis of TCGA data.RegRNA2 and miRanda databases were used to predict the potential target miRNAs of LncRNA SLC16A1-AS1.The expression of LncRNA SLC16A1-AS1 in triple negative breast canceRtissues and cell lines(BT549,BT20,MDA-MB-231,MDA-MB-468)was detected by qRT-PCR.The binding ability of LncRNA SLC16A1-AS1 and miR-182 was verified by double luciferase activity assay and RNA immunoprecipitation test.Cell counting Kit-8 and colony formation test were used to detect the changes of BT549 cells proliferation ability in different treatment groups.Results The results of bioinformatics analysis showed that the expression level of LncRNA SLC16A1-AS1 in TNBC tissues was significantly loweRthan that in normal tissues(P<0.001),which was correlated with good prognosis(P<0.001).There were biding sites between LncRNA SLC16A1-AS1 and miR-182.The expression level of LncRNA SLC16A1-AS1 in 63 TNBC tissues was significantly loweRthan that in adjacent tissues(7.94%vs 63.49%,P<0.001),and the expression level in TNBC cell lines was also significantly loweRthan that in normal breast epithelial cell line(P<0.01).In BT549 cells,overexpression of miR-182 significantly reduced the fluorescein activity of LncRNA SLC16A1-AS1-WT.RNA immunoprecipitation test showed that AGO2 enriched both of LncRNA SLC16A1-AS1 and miR-182(P<0.01).Compared with the control group,overexpression of LncRNA SLC16A1-AS1 significantly decreased the proliferative activity and the numbeRof colony formation of BT549 cells,and co-expression of LncRNA SLC16A1-AS1 and miR-182 restored the proliferative ability of BT549 cells(P<0.01).Conclusion LncRNA SLC16A1-AS1 is lowly expressed in TNBC and inhibits the proliferation of TNBC cells in vitro by tar

关 键 词：乳腺肿瘤 三阴型乳腺癌 LncRNA SLC16A1-AS1 miR-182 

分 类 号：R737.9[医药卫生—肿瘤]