【摘要】 目的观察RNA结合基序蛋白5-反义链1(RBM5-AS1)在甲状腺癌(TC)细胞中的表达变化,及其对细胞增殖、凋亡、侵袭、迁移能力的影响,并探讨可能机制。方法采用qRT-PCR法检测TC细胞C643、B-CPAP、TPC-1、BHT101、KMH-2、8305C和正常甲状腺细胞Nthy-ori 3-1中的RBM5-AS1,选择表达水平最高的细胞进行后续实验。将TC细胞分别转染NC siRNA(对照组)和RBM5-AS1 siRNA(实验组),通过CCK-8实验检测细胞增殖能力,流式细胞术测算细胞凋亡率,Transwell实验分别检测细胞侵袭、迁移能力,Western blotting法检测雷帕霉素靶蛋白(mTOR)-p70核糖体蛋白S6激酶(p70S6K)/真核翻译起始因子4E结合蛋白1(4E-BP1)信号通路蛋白表达水平。结果 C643、B-CPAP、TPC-1、BHT101、KMH-2、8305C细胞中RBM5-AS1相对表达量均高于Nthy-ori 3-1细胞,其中8305C细胞RBM5-AS1表达量最高(P均<0.05)。与对照组相比,实验组细胞增殖能力减弱,细胞凋亡率升高,侵袭及迁移穿膜细胞数减少,p-mTOR、p70S6K、4E-BP1蛋白相对表达量降低(P均<0.05)。结论 TC细胞中RBM5-AS1表达上调;干扰RBM5-AS1表达抑制了TC细胞的增殖、迁移、侵袭能力,并促进其凋亡,其机制可能与调控mTOR/p70S6K/4E-BP1信号通路有关。更多还原

【Abstract】 Objective To investigate the expression changes of lncRNA RNA-binding motif protein 5 antisense RNA 1(RBM5-AS1) in thyroid carcinoma(TC) cells and its effects on cell proliferation, apoptosis, migration, and invasion of TC cells, and to explore the possible mechanism.Methods The qRT-PCR was used to detect the expression levels of RBM5-AS1 in C643, B-CPAP, TPC-1, BHT101, KMH-2, 8305C and normal thyroid cell line Nthy-ori 3-1. The TC cell line with the highest RBM5-AS1 expression was selected for the subsequent experiments. TC cells were transfected with NC siRNA(control group) and RBM5-AS1 siRNA(experimental group), respectively. CCK-8 was used to detect the proliferation ability of cells. Flow cytometry was used to measure apoptosis rate. Transwell test was used to detect the migration and invasion abilities. Western blotting was used to detect the mTOR/p70S6K/4E-BP1 signaling pathway-related protein.Results Compared with Nthy-ori 3-1 cell line, the expression of RBM5-AS1 was higher in C643, B-CPAP, TPC-1,BHT101, KMH-2, 8305C cell lines, with the highest RBM5-AS1 expression in 8305C cell line(all P< 0. 05). Compared with the control group, the cell proliferation, the number of migration and invasion transmembrane cells, p-mTOR,p70S6K, 4E-BP1 protein expression in the experimental group decreased, and the apoptosis ability was enhanced(all P<0. 05).Conclusions The expression of RBM5-AS1 is up-regulated in TC cells. Down-regulation of RBM5-AS1 expression can inhibit cell proliferation and metastasis but enhance apoptosis, and the mechanism may be related to the regulation of mTOR/p70S6K/4E-BP1 signaling pathway.更多还原