微RNA-152靶向调控内质网脂质Raft关联蛋白1对甲状腺乳头状癌细胞增殖和迁移的影响

Effect of microRNA-152 on proliferation and migration of papillary thyroid carcinoma cells by targeting endoplasmic reticulum lipid raft associated protein 1

目的探讨微RNA-152(miR-152)对甲状腺乳头状癌(PTC)细胞增殖、侵袭和迁移的影响及其作用机制。方法将对数生长期PTC TPC-1细胞随机分为空白对照组、miR-con组(转染miRNA-mimics阴性对照)和miR-152组(转染miR-152-mimics),另取对数生长期正常甲状腺细胞株Nthy-ori 3-1作为Nthy-ori 3-1组。采用实时荧光定量聚合酶链反应法检测各组细胞中miR-152、内质网脂质Raft关联蛋白1(ERLIN1)mRNA表达,四唑盐比色法及平板克隆形成实验检测各组TPC-1细胞增殖能力,流式细胞仪检测各组TPC-1细胞凋亡情况,Transwell实验检测各组TPC-1细胞迁移、侵袭能力,Western blot法检测各组TPC-1细胞中ERLIN1蛋白表达。结果空白对照组与miR-con组TPC-1细胞中miR-152、ERLIN1 mRNA和蛋白相对表达量、TPC-1细胞增殖抑制率、细胞克隆数、细胞凋亡率、细胞迁移数及侵袭数比较差异均无统计学意义(P>0.05)。空白对照组、miR-con组、miR-152组TPC-1细胞中miR-152相对表达量显著低于Nthy-ori 3-1组(P<0.05);miR-152组TPC-1细胞中miR-152相对表达量显著高于空白对照组和miR-con组(P<0.05)。与空白对照组、miR-con组比较,miR-152组TPC-1细胞增殖抑制率、细胞凋亡率显著升高,细胞克隆数、细胞迁移数及侵袭数显著降低(P<0.05)。miR-152组TPC-1细胞中ERLIN1 mRNA及蛋白相对表达量均显著低于空白对照组和miR-con组(P<0.05)。结论过表达miR-152可下调TPC-1细胞中ERLIN1的表达,抑制TPC-1细胞增殖、侵袭和迁移,miR-152可能通过靶向调控ERLIN1参与PTC进展和转移。

Objective To investigate the effect of microRNA(miR)-152 on the proliferation,invasion and migration of papillary thyroid carcinoma(PTC)cells and its mechanism.Methods PTC TPC-1 cells in logarithmic growth phase were randomly divided into blank control group,miR-con group(transfected with miRNA-mimics negative control)and miR-152 group(transfected with miR-152-mimics),and the normal thyroid cells Nthy-ori 3-1 were taken as the Nthy-ori 3-1 group.The expressions of miR-152 and endoplasmic reticulum lipid raft associated protein 1(ERLIN1)mRNA in each group were detected by real-time fluorescence quantitative polymerase chain reaction,the proliferation ability of TPC-1 cells in each group was detected by methyl thiazolyl tetrazolium method and plate colony forming test,the apoptosis of TPC-1 cells in each group was detected by flow cytometry,the cell migration and invasion ability of TPC-1 cells in each group were detected by Transwell test,the expression of ERLIN1 protein of TPC-1 cells in each group was detected by Western blot.Results There was no significant difference in the relative expression of miR-152 and ERLIN1 mRNA and protein,the inhibition rate of TPC-1 cells proliferation,the cell clone number,the rate of apoptosis,the number of cell migration and invasion between the blank con trol group and the miR-con group(P>0.05).The relative expression of miR-152 in the blank control group,miR-con group and miR-152 group was significantly lower than that in the Nthy-ori 3-1 group(P<0.05);the relative expression of miR-152 in the miR-152 group was significantly higher than that in the blank control group and the miR-con group(P<0.05).Compared with the blank control group and miR-con group,the inhibition rate of cell proliferation,apoptosis rate of TPC-1 cells in miR-152 group were significantly increased(P<0.05);and the cell clone number,migration and invasion ability of TPC-1 cells in miR-152 group were significantly decreased(P<0.05).The relative expressions of ERLIN1 mRNA and protein of TPC-1 cells in the miR-152 group were significantly lower than those in the blank control group and the miR-con group(P<0.05).Conclusion Overexpression of miR-152 can down-regulate the expression of ERLIN1 in TPC-1 cells,inhibit the proliferation,invasion and migration of TPC-1 cells.It is suggested that miR-152 may be involved in the progression and metastasis of PTC by targeting ERLIN1.