prospero同源框1下调对人胶质瘤细胞增殖、侵袭的影响

Effect of down-regulation of prospero homeobox 1 on the proliferation and invasion of human glioma cells

目的研究prospero同源框1(PROX1)对人胶质瘤细胞体外增殖、侵袭的影响。方法用定量逆转录聚合酶链反应(qRT-PCR)和Westernblot法筛选高表达PROX1基因的胶质瘤细胞株,用RNA干扰沉默PROX1,将细胞分为对照组、scramble siRNA组、PROX1 siRNA组,通过噻唑蓝(MTT)实验和Transwell细胞侵袭实验,观察PROX1下调对胶质瘤生物学效应的影响。用Westernblot法检测上皮-间充质转化(EMT)标志蛋白兔抗人E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)的表达。结果PROX1在人胶质瘤细胞A172、U87MG、U118MG、SHG-44和U251中蛋白和mRNA的表达水平均高于正常人胶质细胞HEB,且U118MG和U87MG细胞中PROX1蛋白和mRNA表达水平均高于其他人胶质瘤细胞,差异均有统计学意义(P﹤0.05)。在72、96 h,PROX1 siRNA组U118MG和U87MG细胞增殖能力低于scramble siRNA组和对照组,差异均有统计学意义(P﹤0.05)。PROX1 siRNA组U118MG和U87MG细胞侵袭数目分别少于对照组和scramble siRNA组,差异均有统计学意义(P﹤0.05)。PROX1 siRNA组U118MG和U87MG细胞中E-cadherin表达水平均明显高于对照组,N-cadherin和Vimentin表达水平均明显低于对照组,差异均有统计学意义(P﹤0.01)。结论PROX1在胶质瘤细胞中高表达,可促进胶质瘤细胞的增殖,有望成为胶质瘤基因治疗的潜在靶点。

Objective To investigate the effects of prospero homeobox 1(PROX1)on the proliferation and invasion of human glioma cells in vitro.Method quantitative reverse transcription-polymerase chain reaction(qRT-PCR)and Western blot were used to screen glioma cell lines with up-regulation of PROX1 gene,and PROX1 was silenced by RNA interference.The cells were divided into the control group,scramble siRNA group,and PROX1 siRNA group.Through the methyl thiazolyl tetrazolium(MTT)experiment and Transwell assay,the effect of down-regulation of PROX1 on the biological effects of glioma was observed.Western blot was used to detect the expression of rabbit anti-human E-cad-herin,N-cadherin,and Vimentin,which were the biomarkers of the epithelial-mesenchymal transition(EMT)process.Re-sult Protein and mRNA expression levels of PROX1 in human glioma cells A172,U87MG,U118MG,SHG-44,and U251 were higher than the normal human glial cell line HEB,and the expression levels in U118MG and U87MG cells were higher than the other human glioma cell lines,the differences were statistically significant(P<0.05).At 72 and 96 h,the proliferation competence of U118MG and U87MG cells in the PROX1 siRNA group were lower than those of the scramble siRNA group and the control group,the differences were statistically significant(P<0.05).The number of U118MG and U87MG invasive cells in the PROX1 siRNA group were less than those of the control and scramble siRNA groups,and the differences were statistically significant(P<0.05).The expression levels of E-cadherin in U118MG and U87MG cells in the PROX1 siRNA group were significantly higher than those in the control group,and the expression levels of N-cadherin and Vimentin were significantly lower than those in the control group,the differences were statisti-cally significant(P<0.01).Conclusion PROX1 is up-regulation in glioma cells,which could promote glioma cells’pro-liferation and is expected to become a promising and potential target for glioma gene therapy.