论文部分内容阅读
该研究旨在探讨乳酸脱氢酶A(lactate dehydrogenase A,LDHA)基因对脑胶质瘤细胞上皮细胞–间充质转化(epithelial-mesenchymal transition,EMT)以及细胞生长的影响。用sh-LDHA质粒转染脑胶质瘤细胞,比色法检测细胞内LDHA活性,流式细胞术检测细胞凋亡率,CCK-8法和克隆形成实验分析细胞增殖能力,划痕实验和Transwell实验检测细胞迁移能力,q-PCR检测LDHAm RNA水平,以及Western blot检测LDHA和EMT相关蛋白质水平。结果表明,转染sh-LDHA质粒能显著降低LDHA m RNA和蛋白质水平。同时,敲低LDHA诱导U87MG细胞发生凋亡,LDHA活性降至原来水平的50%U87MG细胞的增殖和迁移能力低于sh-EGFP组(P<0.05)。另外,敲低LDHA抑制U87MG细胞的EMT过程。而在LDHA表达水平相对较低的U251MG和SW1783细胞中,干扰LDHA对细胞生长和EMT没有显著影响。该研究结果说明,LDHA在脑胶质瘤细胞的EMT过程和生长中发挥了重要作用。
This study aimed to investigate the effect of lactate dehydrogenase A (LDHA) gene on epithelial-mesenchymal transition (EMT) and cell growth in glioma cells. Glioma cells were transfected with sh-LDHA plasmid, LDHA activity was detected by colorimetric assay, apoptosis rate was detected by flow cytometry, CCK-8 assay and clonogenic assay were used to analyze cell proliferation, scratch test and Transwell The migration ability of cells was detected by qRT-PCR and LDHAm RNA was detected by q-PCR. Western blot was used to detect the protein levels of LDHA and EMT. The results showed that transfection with sh-LDHA plasmid can significantly reduce LDHA m RNA and protein levels. Meanwhile, the knockdown of LDHA induced apoptosis in U87MG cells, and the LDHA activity was reduced to 50% of the original level. The proliferation and migration of U87MG cells were lower than that of sh-EGFP group (P <0.05). In addition, knockdown of LDHA inhibited the EMT process in U87MG cells. However, in U251MG and SW1783 cells with relatively low levels of LDHA, interference with LDHA had no significant effect on cell growth and EMT. The results of this study indicate that LDHA plays an important role in the EMT process and growth of glioma cells.