miR-223-3p通过靶向TGFBR3促进LncRNA ADAMTS9-AS2表达上调抑制肺癌细胞的增殖和迁移作用

miR-223-3p promotes the up-regulation of LncRNA ADAMTS9-AS2 expression by targeting TGFBR3 to inhibit the proliferation and migration of lung cancer cells

目的:探讨miR-223-3p通过靶向转化生长因子-βⅢ型受体(TGFBR3)促进长链非编码RNA(LncRNA)ADAMTS9-AS2表达上调抑制肺癌细胞增殖和迁移的作用及可能机制。方法:采用RT-PCR检测肺癌H1299细胞中miR-223-3p和TGFBR3 mRNA表达水平,Western blotting检测TGFBR3的蛋白表达水平,RT-qPCR检测LncRNA ADAMTS9-AS2的表达水平,CCK-8检测细胞增殖能力,Transwell细胞迁移实验和划痕实验检测细胞迁移能力。采用脂质体转染miR-223-3p模拟物(过表达组)、抑制剂(抑制组)、对照质粒(对照组)于H1299细胞中,比较各组转染前后miR-223-3p、TGFBR3、LncRNA ADAMTS9-AS2表达水平、细胞增殖能力、细胞迁移能力。结果:与转染前比较,过表达组转染后的H1299细胞中miR-223-3p、LncRNA ADAMTS9-AS2表达水平均升高(P<0.05),TGFBR3蛋白和mRNA表达水平均降低(P<0.01和P<0.05),细胞增殖和迁移能力下降(P<0.05);抑制组转染后的细胞中miR-223-3p和LncRNA ADAMTS9-AS2表达水平均降低(P<0.05),TGFBR3蛋白和mRNA表达水平均升高(P<0.01和P<0.05),细胞增殖和迁移能力均增加(P<0.05)。转染72 h后,TGFBR3蛋白表达水平及mRNA的表达水平细胞增殖与迁移能力:抑制组>观察组>过表达组(P<0.01)。3组miR-223-3p、LncRNA ADAMTS9-AS2 mRNA表达水平逐渐降低,抑制组<对照组<过表达组(P<0.01)。结论:miR-223-3p抑制肺癌H1299细胞的增殖和迁移,靶向下调TGFBR3和上调LncRNA ADAMTS9-AS2表达可能为其作用机制。

Objective:To investigate the effect of miR-223-3p on the up-regulation of long non-coding RNA(LncRNA)ADAMTS9-AS2 by targeting transforming growth factor-βtypeⅢreceptor(TGFBR3)to inhibit the proliferation and migration of lung cancer cells.Methods:The mRNA expression levels of miR-223-3p and TGFBR3 in lung cancer H1299 cells were detected by RT-PCR,the protein expression level of TGFBR3 was detected by Western blotting,the expression level of LncRNA ADAMTS9-AS2 was detected by RT-qPCR,the cell proliferation ability was detected by CCK-8,and the cell migration ability was detected by Transwell migration assay and scratch assay.MiR-223-3p mimic(overexpression group),inhibitor(inhibition group)and control plasmid(control group)were transfected into H1299 cells by liposome.The expression levels of miR-223-3p,TGFBR3,LncRNA ADAMTS9-AS2,cell proliferation and cell migration were compared between before and after transfection.Results:Compared with before transfection,the expression levels of miR-223-3p mRNA and LncRNA ADAMTS9-AS2 in H1299 cells after transfection in the overexpression group increased(P<0.05),the expression levels of TGFBR3 protein and mRNA decreased(P<0.01 and P<0.05),and the cell proliferation and migration ability decreased(P<0.05);the expression levels of miR-223-3p mRNA and LncRNA ADAMTS9-AS2 in the inhibition group decreased(P<0.05),the expression levels of TGFBR3 protein and mRNA increased(P<0.01 and P<0.05),and the cell proliferation and migration ability increased(P<0.05).After 72 h of transfection,the expression level of TGFBR3 protein and mRNA,and the cell proliferation and migration ability were as follows:inhibition group>observation group>overexpression group(P<0.01).The expression levels of miR-233-3p and LncRNA ADAMTS9-AS2 mRNA gradually decreased,Which were as follows:inhibition group<control group<overexpression group(P<0.01).Conclusions:MiR-223-3p inhibits the proliferation and migration of lung cancer H1299 cells,the mechanism of which may involve the targeted down-regulation of TGFBR3 and up-regulation of LncRNA ADAMTS9-AS2 expression.