外源性干预VEGF-C表达对肺癌细胞生物学行为的影响及机制

Effect of exogenous VEGF-C on biological behavior of lung cancer cells and its mechanism

目的探讨外源性血管内皮生长因子-C(VEGF-C)对肺癌细胞生物学行为的影响及机制。方法将A549细胞分为BK组、VC组及SC组,BK组不转染,VC组及SC组均于EP管中加入200μL转染液和4μL脂质体,VC组及SC组分别加入5μg NCmimics、5μg miR-21 mimics。转染72 h后,采用免疫印迹法检测A549细胞中TCAB1蛋白表达;MTT法检测A549细胞生存率;Transwell小室检测A549细胞侵袭情况;流式细胞仪检测A549细胞凋亡率。结果SC组A549细胞中VEGF-C mRNA表达低于BK组、VC组(P均<0.05),转染成功。BK组、VC组中VEGF-C mRNA表达相似(P>0.05)。干预12、24、48、72 h时,BK组与VC组A549细胞生存率比较差异无统计学意义(P均>0.05);SC组A549细胞生存率低于BK组、VC组,组间比较差异有统计学意义(P均<0.05)。BK组与VC组A549细胞侵袭细胞数目比较差异无统计学意义(P>0.05),SC组A549细胞侵袭细胞数目低于BK组、VC组,三组比较差异有统计学意义(F=24.383,P<0.001)。BK组与VC组A549细胞凋亡率比较差异无统计学意义(P>0.05);SC组A549细胞凋亡率高于BK组、VC组,三组比较差异有统计学意义(F=125.326,P<0.05)。BK组、VC组A549细胞中TCAB1蛋白表达分别为2.03±0.21、1.98±0.20,两组比较差异无统计学意义(P>0.05),SC组A549细胞中TCAB1蛋白表达为1.35±0.12,低于BK组、VC组(P均<0.05)。结论外源性降低VEGF-C表达可降低A549细胞存活率及侵袭能力,提高细胞凋亡率;其机制可能与抑制TCAB1蛋白表达有关。

Objective To investigate the effect of exogenous vascular endothelial growth factor-C(VEGF-C)on the biological behavior of lung cancer cells and its mechanism.Methods We divided A549 cells into BK group,VC group,and SC group.A549 cells in the BK group were not transfected;the A549 cells in the VC group and SC group were added with 200μL transfected liquid and 4μL lipids into the EP tubes,and then A549 cells in the VC group and SC group were added with 5μg NC mimics and MIR-21 mimics,respectively.The expression of TCAB1 protein in A549 cells was detected by Western blotting.The survival rate of A549 cells was detected by MTT.Transwell chamber was used to detect the inva⁃sion of A549 cells.The apoptosis rate of A549 cells was detected by flow cytometry.Results The mRNA expression of VEGF-C in lung cancer tissues was lower in the SC group than in the BK group and the VC group(P<0.05),which indi⁃cated that the transfection was successful;the expression levels of VEGF-C mRNA in the BK group and the VC group were similar(P>0.05).There was no significant difference in the survival rate of A549 cells between the BK group and the VC group at 12,24,48,and 72 hours of intervention(P>0.05);the survival rate of A549 cells in the SC group was lower than those of the BK group and the VC group,and there was significant difference between the groups(P<0.05).There was no significant difference in the number of invasion cells between the BK group and the VC group(P>0.05).The num⁃ber of invasion cells in the SC group was significantly lower than those of the BK group and the VC group;there were signif⁃icant differences between the three groups(F=24.383,P<0.001);there was no difference in the apoptosis rate between the BK group and the VC group(P>0.05).The apoptosis rate of A549 cells in the SC group was significantly higher than those of the BK group and the VC group;there were differences among the three groups(F=125.326,P<0.05).No signif⁃icant difference was found in the expression of TCAB1 protein in the lung cancer tissues between the BK group and VC group(2.03±0.21 versus 1.98±0.20)(P>0.05),and the expression level of TCAB1 protein in A549 cells of the SC group(1.35±0.12)was lower than those of the BK group,VC group(both P<0.05).Conclusion Reducing the level of exogenous VEGF-C can reduce the survival rate and invasion ability of A549 cells,and increase the apoptosis rate;its mechanism may be related to the inhibition of TCAB1 protein.