长链非编码RNA RP11-385J1.2通过靶向微小RNA-370-3p调控甲状腺癌细胞增殖、侵袭及凋亡的作用机制

Mechanism of long non-coding RNA RP11-385J1.2 regulating the proliferation,invasion and apoptosis of thyroid cancer cells by targeting microRNA-370-3p

目的探讨长链非编码RNA(lncRNA)RP11-385J1.2对甲状腺癌细胞增殖、侵袭和凋亡的影响。方法采用实时荧光聚合酶链反应(PCR)检测人甲状腺癌细胞株SW579、FTC-133、TPC-1、K1和正常甲状腺细胞Nthyori 3-1中RP11-385J1.2和微小RNA(miRNA)-370-3p的表达情况。采用抑制剂si-RP11-385J1.2和(或)anti-miRNA-370-3p转染SW579细胞,噻唑蓝(MTT)法检测细胞增殖,Transwell实验检测细胞侵袭,流式细胞术检测细胞凋亡,蛋白质印记法(Western blot)检测凋亡相关蛋白B细胞淋巴瘤/白血病-2(Bcl-2)和Bax的表达情况,双荧光素酶报告系统验证RP11-385J1.2和miRNA-370-3p的靶向关系。结果人甲状腺癌细胞株SW579、FTC-133、TPC-1、K1中RP11-385J1.2的表达水平均高于正常甲状腺细胞Nthy-ori 3-1,miRNA-370-3p的表达水平均低于正常甲状腺细胞Nthy-ori 3-1,差异均有统计学意义(P﹤0.05)。敲减RP11-385J1.2可抑制SW579细胞增殖和侵袭并促进细胞凋亡,RP11-385J1.2能够靶向负调控miRNA-370-3p的表达,下调miRNA-370-3p的表达可逆转敲减RP11-385J1.2对SW579细胞增殖、侵袭和凋亡的影响。结论lncRNA RP11-385J1.2通过靶向下调miRNA-370-3p的表达促进甲状腺癌SW579细胞增殖和侵袭并抑制细胞凋亡。RP11-385J1.2和miRNA-370-3p是甲状腺癌的潜在分子靶点。

Objective To investigate the effect of long non-coding RNA(lncRNA)RP11-385J1.2 on the proliferation,invasion and apoptosis of thyroid cancer cells.Method The expression of RP11-385J1.2 and microRNA(miRNA)-370-3p in thyroid cancer cell lines SW579,FTC-133,TPC-1 and K1 as well as in normal thyroid cells Nthy-ori 3-1 were detected by real-time polymerase chain reaction.The SW579 cells were transfected with inhibitors of si-RP11-385J1.2 and/or anti-miRNA-370-3p,respectively.Thiazolyl blue tetrazolium bromide(MTT)was used to detect cell proliferation,transwell was performed to determine cell invasion,and flow cytometry to assess cell apoptosis;besides,the expression of apoptosis-related proteins B-cell lymphoma/leukemia-2(Bcl-2)and Bax were detected by Western blot;dual-luciferase reporter assay system was applied to investigate the relationship between RP11-385J1.2 and miRNA-370-3p.Result Compared with normal Nthy-ori 3-1 cells,the expression level of RP11-385J1.2 in thyroid cancer cell lines SW579,FTC-133,TPC-1 and K1 were increased significantly,and the expression of miRNA-370-3p was decreased remarkably(P<0.05).Knockdown of RP11-385J1.2 inhibited the proliferation and invasion and promoted cell apoptosis of SW579 cells.RP11-385J1.2 regulated the expression of miRNA-370-3p in a negative and targeted manner.Downregulating miRNA-370-3p reversed the effects of RP11-385J1.2 knockdown on proliferation,invasion and apoptosis of SW579 cells.Conclusion lncRNA RP11-385J1.2 promotes proliferation and invasion while inhibits apoptosis of thyroid cancer SW579 cells by targeted downregulation of miRNA-370-3p.RP11-385J1.2 and miRNA-370-3p are potential molecular targets for thyroid cancer.