miR-143-3p靶向调控HMGA2影响三阴性乳腺癌细胞体外转移活性

Effect of miR-143-3p on the metastatic activity of triple-negative breast cancer cells in vitro by targeting HMGA2

[目的]探究miR-143-3p与HMGA2在对三阴性乳腺癌细胞的生物学过程的影响。[方法]采用实时荧光定量PCR方法比较不同组织中miR-143-3p的表达水平,采用免疫组化法检测不同组织中HMGA2的表达情况。将MDA-MB-231细胞株分为4组:miR NC组、miR-143-3p mimic组、inhibitor NC组和HMGA2 inhibitor组。蛋白免疫印迹检测HMGA2蛋白的表达水平;采用细胞集落形成实验研究细胞的生长能力;Transwell法研究细胞的侵袭能力;细胞划痕实验分析细胞迁移能力;流式细胞术检测细胞凋亡率;应用生物信息学方法预测miR-143-3p与HMGA2的结合位点;双荧光素酶报告基因实验分析miR-143-3p与HMGA2的靶向关系。[结果]三阴性乳腺癌组织中miR-143-3p表达水平降低(0.98±0.02 vs 0.46±0.03),HMGA2增加(P<0.05);miR-143-3p能够靶向抑制HMGA2的活性(1.09±0.02 vs 0.32±0.01,P<0.05);miR-143-3p mimic或HMGA2 inhibitor均能抑制MDA-MB-231细胞的生长、迁移和侵袭能力,并能促进MDA-MB-231细胞凋亡(4.69%±0.02%vs 16.32%±1.01%vs 4.17%±0.05%vs 15.22%±0.03%;P<0.05)。[结论]HMGA2是miR-143-3p的直接靶点,miR-143-3p可能通过靶向调控HMGA2抑制三阴性乳腺癌细胞的体外转移活性,miR-143-3p与HMGA2的关系有望成为三阴性乳腺癌治疗的关键靶点。

[Objective]To investigate the effect of miR-143-3p and HMGA2 on the biological process of triple-negative breast cancer cells.[Method]The expression levels of miR-143-3p in different tissues were compared by real-time fluorescence quantitative PCR,and the expression of HMGA2 in different tissues was detected by immunohistochemistry.The MDA-MB-231 cell line was divided into four groups:miR NC group,miR-143-3p mimic group,inhibitor NC group and HMGA2 inhibitor group.Protein immunoblotting was used to detect the protein expression level of HMGA2;cell colony formation assay was used to study the growth ability of cells;Transwell assay was used to study the invasion ability of cells;cell scoring assay was used to analyze the migration ability of cells;flow cytometry was used to detect the apoptosis rate;bioinformatics method was applied to predict the binding site of miR-143-3p and HMGA2;dual luciferase reporter gene assay was performed to analyze the targeting relationship between miR-143-3p and HMGA2.[Result] The expression level of miR-143-3p was decreased(0.98±0.02 vs 0.46±0.03)and HMGA2 was increased(P<0.05)in triple-negative breast cancer tissues.miR-143-3p could inhibit the activity of HMGA2(1.09±0.02 us 0.32±0.01,P<0.05).miR-143-3p mimic or HMGA2 inhibitor could inhibit the growth,migration and invasion of MDA-MB-231 cells,the apoptosis of MDA-MB-231 cells(4.69%±0.02%vs 16.32%±1.01%vs 4.17%±0.05%us 15.22%±0.03%;P<0.05).[Conclusion]HMGA2 is a direct target of miR-143-3p.miR-143-3p may inhibit the metastatic activity of triple-negative breast cancer cells in vitro by targeting and regulating HMGA2,and the relationship between miR-143-3p and HMGA2 is expected to be a key target for the treatment of triple-negative breast cancer.