长链非编码RNA UCA1在TGF-β介导的胃癌细胞EMT中的作用机制

Mechanism of long non-coding RNA UCA1 in TGF-β-mediated epithelial mesenchymal transformation of gastric cancer cells

目的探讨长链非编码RNA(lncRNA)尿路上皮癌相关基因1(UCA1)在转化生长因子β(TGF-β)介导的胃癌细胞上皮间充质转化(EMT)中的作用机制。方法用10 ng/mL人重组TGF-β1处理胃癌细胞系MNK-45及SGC-7901,Transwell实验检测细胞侵袭能力,荧光定量PCR检测Slug、锌指E盒结合同源框1(ZEB1)、波形蛋白和上皮型钙黏蛋白(E-cadherin)的表达水平;用TGF-β处理细胞后转染lncRNA UCA1小干扰RNA(siUCA1),检测上述指标。在MNK-45细胞中转染微小RNA-204(miR-204)模拟物、miR-204模拟物+lncRNA UCA1过表达质粒5 ng、miR-204模拟物+lncRNA UCA1过表达质粒10 ng后,检测ZEB1的表达水平。用TGF-β、TGF-β+miR-204模拟物、TGF-β+miR-204模拟物+lncRNA UCA1过表达质粒处理MNK-45细胞,检测Slug、ZEB1、波形蛋白和E-cadherin的表达水平。结果与对照组比较,TGF-β处理组穿膜细胞数增多,Slug、ZEB1、波形蛋白和lncRNA UCA1表达增加,E-cadherin表达减少(P<0.01)。与TGF-β处理组比较,TGF-β+siUCA1组穿膜细胞数减少,Slug、ZEB1、波形蛋白表达减少,E-cadherin表达增加(P<0.01)。转染miR-204模拟物后可抑制ZEB1的表达(P<0.01),同时转染lncRNA UCA1过表达质粒(5 ng和10 ng)后,对ZEB1的抑制作用消失。与TGF-β处理组比较,TGF-β+miR-204模拟物组Slug、ZEB1和波形蛋白表达减少,E-cadherin表达增加(P<0.01),而同时转染lncRNA UCA1过表达质粒(5 ng)后,可阻断TGF-β+miR-204模拟物的作用(P<0.01)。结论LncRNA UCA1能够通过miR-204/ZEB1影响TGF-β对胃癌细胞EMT的促进作用。

Objective To investigate the mechanism of long non-coding RNA UCA1 in transforming growth factor(TGF)-β-mediated epithelial mesenchymal transformation(EMT)of gastric cancer cells.Methods Gastric cancer cell lines MNK-45 and SGC-7901 were treated with restructured TGF-β110 ng/mL.Cell invasion was measured by Transwell assay and the expressions of Slug,Zinc finger E-box binding homeobox1(ZEB1),vimentin and E-cadherin were detected by fluorescent quantitative PCR.The above indicators were detected after transfected with siUCA1.MNK-45 cells were transfected with miR-204 mimics,miR-204 mimics+lncRNA UCA1 overexpressed plasmids 5 ng and miR-204 mimics+lncRNA UCA1 overexpressed plasmids 10 ng.The expression of ZEB1 was detected.MNK-45 cells were treated with TGF-β,TGF-β+miR-204 mimics and TGF-β+miR-204 mimics+lncRNA UCA1 overexpressed plasmids.The expressions of Slug,ZEB1,vimentin and E-cadherin were detected.Results The number of transmembrane cells was increased,the expressions of Slug,ZEB1,vimentin and lncRNA UCA1 were higher and E-cadherin was lower after treated with TGF-β(P<0.01).The number of transmembrane cells was decreased,the expressions of Slug,ZEB1 and vimentin were lower and E-cadherin was higher after treated with TGF-β+siUCA1 than treated with TGF-β(P<0.01).Transfection with miR-204 mimics inhibited the expression of ZEB1,and the inhibitory effect disapeared after transfected with miR-204 mimics+lncRNA UCA1 overexpressed plasmids(5 ng and 10 ng)(P<0.01).The expressions of Slug,ZEB1 and vimentin were lower and E-cadherin was higher after treated with TGF-β+miR-204 mimics than treated with TGF-β,and the effect was blocked after treated with TGF-β+miR-204 mimics+lncRNA UCA1 overexpressed plasmids(5 ng)(P<0.01).Conclusion Long non-coding RNA UCA1 can influence the effect of TGF-βon EMT in gastric cancer cells through miR-204/ZEB1.