肿瘤坏死因子相关诱导凋亡配体联合蛋白酶抑制剂MG132对人肺腺癌细胞GLC-82血小板衍生生长因子受体α-磷脂酶Cγ1-蛋白激酶Cα信号转导的研究

Platelet derived growth factor receptor α-phospholipase Cγ1-protein kinase Cα signal transductionin in human lung cancer cell GLC-82 for combination of the drugs tumor necrosis factor-related apoptosis-inducing ligand and proteasome inhibitor MG132

目的 观察肿瘤坏死因子相关诱导凋亡配体（TRAIL）联合蛋白酶抑制剂MG132对人肺腺癌细胞GLC-82增殖的抑制和血小板衍生生长因子受体α（PDGFRα）-磷脂酶Cγ1（PLCγ1）-蛋白激酶Cα（PKCα）信号转导通路的影响.方法 用噻唑蓝（MTT）法检测TRAIL联合蛋白酶抑制剂MG132对GLC-82细胞生长的抑制作用;采用流式细胞仪技术检测TRAIL联合蛋白酶抑制剂MG132对GLC-82细胞凋亡的影响,采用链霉菌抗生物素蛋白-过氧化物酶（SP）免疫组织化学法检测TRAIL联合蛋白酶抑制剂MG132相关蛋白PDGFRα、PLCγ1、PKCα的表达,Western blot测定TRAIL联合蛋白酶抑制剂MG132对GLC-82细胞PDGFRα、PLCγ1、PKCα蛋白表达的影响.结果 MTI法显示TRAIL联合蛋白酶抑制剂MG132作用GLC-82细胞株后,抑制率较单独应用TRAIL和MG132均显著增高,抑制率分别为（34.92±3.95）％、（33.17±1.78）％和（76.96±1.28）％（P＜0.05）;流式细胞仪细胞凋亡检测结果显示,TRAIL联合蛋白酶抑制剂MG132作用GLC-82细胞48 h后TRAIL联合蛋白酶抑制剂MG132能明显诱导GLC-82细胞凋亡（P＜0.05）,免疫组织化学检测结果显示TRAIL联合蛋白酶抑制剂MG132作用GLC-82细胞株后能使PDGFRα、PLCγ1、PKCα蛋白表达降低,Western blot结果显示TRAIL联合蛋白酶抑制剂MG132明显抑制PDGFRα、PLCγ1、PKCα蛋白的表达.结论 TRAIL联合蛋白酶抑制剂MG132对人肺腺癌GLC-82细胞的生长有明显的抑制作用,能降低GLC-82细胞PDGFRα-PLCγ1-PKCα信号通路各蛋白表达的水平,肺腺癌的发病和PDGFRα-PLCγ1-PKCα转导通路信号增强关系密切.

Objective To tumor necrosis factor-related apoptosis-inducing ligand （TRAIL） combined with a protease inhibitor MG132 observed in human lung adenocarcinoma cell line GLC-82 inhibition of proliferation and platelet derived growth factor receptor α （PDGFRα）-phospholipase Cγ1 （PLCγ1）-protein kinase Cα （PKCα） signal transduction pathway.Methods Methyl thiazol tetrazolium （MTT） assay inhibition of TRAIL combined with a protease inhibitor MG132 on GLC-82 cell growth detection; using flow cytometry to detect the effects of TRAIL combined with a protease inhibitor MG132 on GLC-82 cell apoptosis,using SP immunohistochemical detection of protease inhibitors MG132 joint expression of TRAIL-associated protein PDGFRα,PLCγ1,PKCα＇ s,Western blotting to determine the effect of protease inhibitors MG132 TRAIL combined for GLC-82 cells PDGFRα,PLCγ1,PKCα protein expression.Results MTT assay inhibition of TRAIL combined with a protease inhibitor MG132 on GLC-82 cell growth detection.The inhibitory rates were （34.92 ± 3.95） %,（33.17 ± 1.78） % and （76.96 ± 1.28） %,respectively; using flow cytometry to detect the effects of TRAIL combined with a protease inhibitor MG132 on GLC-82 cell apoptosis.Apoptosis rates of the four group were 6.78%,38.72%,45.60% and 91.74%,respectively.Using SP immunohistochemical detection of protease inhibitors MG132 joint expression of TRAIL-associated protein PDGFRα,PLCγ1,PKCα＆#39;s,Western blotting to determine the effect of protease inhibitors MG132 TRAIL combined for GLC-82 cells PDGFRα,PLCγ1,PKCα protein expression.Conclusion TRAIL combined protease inhibitor MG132 on human lung adenocarcinoma GLC-82 cells significantly inhibited the growth,can reduce the level of GLC-82 cells PDGFRα-PLCγ1 -PKCα each protein signaling pathway,the incidence of lung cancer and PDGFRα-PLCγ1-PKCα enhanced signal transduction pathways are closely related.