pshRNA-MDRla联合TRAIL基因对胃癌细胞株SGC7901/VCR作用及其作用机制的实验研究  

Experimental study on the effects of pshRNA-MDRla combined with pBUD-TRAIL in inducing SGC7901/VCR cell apoptosis and proliferation and their mechanism

作　　者：袁媛[1] 周洲[1] 周静[1] 姜政[1] 黄爱龙[2] 王丕龙[1] 

机构地区：[1]重庆医科大学附属第一医院消化科,重庆400016 [2]重庆医科大学病毒性肝炎研究所,重庆400010

出　　处：《重庆医科大学学报》2010年第11期1646-1649,共4页Journal of Chongqing Medical University

摘　　要：目的:探讨pshRNA-MDRla联合pBUD-TRAIL,在体外对胃癌细胞SGC7901/VCR增殖和凋亡的作用及机制,为肿瘤的基因治疗奠定基础。方法:以人胃癌SGC7901/VCR细胞株为研究对象,将pshRNA-MDRla与pBUD-TRAIL共同转染胃癌SGC7901/VCR细胞并在VCR中培养,同时与pshRNA-MDRla组、pBUD-TRAIL组、SGC7901/VCR和空质粒组相比较,以MTT法检测细胞增殖抑制率、以FCM检测细胞凋亡率以及TUNEL法检测细胞凋亡等,观察联合应用组对肿瘤细胞增殖的抑制作用。结果:MTT法显示:对照组(pBUD-TRAIL、pshRNA-MDRla、空质粒和SGC7901/VCR组)胃癌细胞的生长抑制率分别为45%、30%、10%和10%,联合实验组抑制率为70%;FCM检测显示:对照组胃癌细胞的凋亡率分别为25.96%、10.27%、5.96%和4.25%,联合实验组凋亡率为41.23%;与对照组相比,联合实验组抑制率和凋亡率都具有显著性差异;TUNEL检测结果显示:联合组与其他对照组相比较,细胞核明显棕黄色深染,且阳性染色细胞数目明显增多,表明pBUD-TRAIL与pshRNA-MDRla联合作用,凋亡细胞明显增加。结论:pshRNA-MDRla联合pBUD-TRAIL可显著抑制肿瘤细胞的增殖和促进肿瘤细胞的凋亡,推测与pshRNA-MDRla阻止TRAIL外排有关,从而增强TRAIL促进肿瘤细胞凋亡的作用,两者具有协同作用。Objective：To investigate the effects of pshRNA-MDRla combined with TRAIL in treating gastric cancer cell lines SGC7901/VCR and understand their mechanism,in order to lay experimental and theoretical foundation for clinical application of gene tumor therapy.Methods：SGC7901/VCR cells transfected with pBUD-TRAIL and pshRNA-MDRla simultaneously were cultivated with 1mg/L concentration of VCR,and compared with SGC7901/VCR cell transfected with pshRNA-MDRla,pBUD-TRAIL,pBUD groups and SGC7901/VCR group,respectively.The inhibition of cell proliferation was examined by methylthiazolyl tetrazolium（MTT） method,and cell apoptosis was detected by TUNEL method and flow cytometry using PI staining analysis.Results：The inhibition rates in pshRNA-MDRla,pBUD-TRAIL,pBUD and SGC7901/VCR groups were respectively 45%,30%,10%,and 10% by MTT.The apoptosis rates in pshRNA-MDRla,pBUD-TRAIL,pBUD and SGC7901/VCR groups were respectively 25.96%,10.27%,5.96%,and 4.25% by FCM.Compared with control group,the inhibition rate of the proliferation and apoptosis rate of SGC7901/VCR cells in pshRNA-MDRla combinated with TRAIL group was higher（70%,41.23% respectively）;Karyopycnosis,nuclear chromosomal condensation and segmentation were observed by TUNEL.pshRNA-MDRla combinated with TRAIL group could significantly induce the apoptosis of tumor.Conclusion：pshRNA-MDRla combined with TRAIL could inhibit the proliferation and induce apoptosis of SGC7901/VCR cells,which could be related with the interfering effects of pshRNA-MDRla by prohibiting TRAIL excretion,so as to intensify the effect of TRAIL in promoting tumor cell apoptosis and have a synergistic action with TRAIL.

关 键 词：pshRNA-MDRla 肿瘤坏死因子相关的凋亡诱导配体 增殖 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]