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该文探讨了TAR RNA结合蛋白2(TAR RNA binding protein 2,TRBP2)基因对肺腺癌A549细胞增殖、迁移及侵袭的影响及可能机制。构建TRBP2慢病毒过表达载体,以不同感染复数(MOI)感染A549细胞,根据绿色荧光强度选择最适MOI值。荧光定量PCR(FQ-PCR)检测TRBP2、MMP-2(matrix metalloproteinase-2)、PKR(double-stranded RNA-dependent protein kinase)m RNA的表达量。免疫蛋白印记(Western blot)法检测TRBP2、MMP-2、PKR、p-PKR的表达。采用MTT法、平板克隆实验检测TRBP2基因对A549细胞增殖的影响,Transwell迁移及侵袭实验检测细胞迁移和侵袭能力,黏附实验检测同种细胞和异种细胞间黏附情况。结果发现,成功构建了TRBP2基因过表达的A549细胞株,与空载体组和对照组比较,TRBP2基因过表达组细胞侵袭、迁移能力明显增强(P<0.05),同种细胞间黏附力减弱(P<0.05)而异种细胞间黏附力增强(P<0.05),增殖速度加快(P<0.05)及克隆形成率增加(P<0.05)。此外,MMP-2、TRBP2蛋白及m RNA表达量明显升高(P<0.05);p-PKR蛋白表达量降低(P<0.05);PKR蛋白及m RNA表达量无差异。对照组与空载体组之间比较,以上各项指标均没有明显差异(P>0.05)。该研究表明,过表达TRBP2基因可能通过促进MMP-2的表达同时抑制PKR磷酸化来促进肺腺癌A549细胞增殖、迁移及侵袭。
This article explores the effect of TAR RNA binding protein 2 (TRBP2) gene on the proliferation, migration and invasion of lung adenocarcinoma A549 cells and its possible mechanism. The TRBP2 lentivirus overexpression vector was constructed and A549 cells were infected with different multiplicity of infection (MOI). The optimal MOI was selected according to the green fluorescent intensity. Fluorescent quantitative PCR (FQ-PCR) was used to detect the expression of TRBP2, matrix metalloproteinase-2 (MMP-2) and double-stranded RNA-dependent protein kinase (mRR) The expressions of TRBP2, MMP-2, PKR and p-PKR were detected by Western blot. The effect of TRBP2 gene on the proliferation of A549 cells was detected by MTT method and plate clone assay. The migration and invasion ability of TRBP2 gene was detected by Transwell migration and invasion assay. The adhesion assay was used to detect the adhesion between the same cells and the dissimilar cells. The results showed that the TRBP2 gene overexpression A549 cell line was successfully constructed. Compared with the empty vector group and the control group, the TRBP2 overexpression group significantly increased the invasion and migration (P <0.05) P <0.05), and the intercellular adhesion was enhanced (P <0.05), the proliferation rate was accelerated (P <0.05) and the colony formation rate was increased (P <0.05). In addition, the expression of MMP-2, TRBP2 protein and m RNA were significantly increased (P <0.05), the expression of p-PKR protein was decreased (P <0.05), and the expression of PKR protein and m RNA was no difference. There was no significant difference between the control group and the empty vector group (P> 0.05). The study shows that overexpression of TRBP2 may promote the proliferation, migration and invasion of lung adenocarcinoma A549 cells by promoting the expression of MMP-2 and inhibiting the phosphorylation of PKR.