肝细胞生长因子基因转染对人淋巴瘤细胞凋亡的影响  

The Effect of Hepatocyte Growth Factor Gene Transfection on the Apoptosis of Human Lymphoma Cell Line Raji Cells

作　　者：沈蓉蓉[1] 郑筱娇[1] 岑东[1,2] 赵行[1] 滑世轩[1] 裴仁治 吕建新[1] 涂植光[4] 

机构地区：[1]温州医学院浙江省医学遗传学重点实验室,温州325027 [2]浙江省鄞州疾病预防控制中心,宁波315100 [3]浙江省鄞州人民医院血液科,宁波315040 [4]重庆医科大学医学检验系教育部"临床检验诊断学"重点实验室,重庆400016

出　　处：《中国细胞生物学学报》2012年第4期325-331,共7页Chinese Journal of Cell Biology

基　　金：浙江省医药卫生科技项目(No.2007A175);宁波市科技计划项目(No.2007C10065;No.2010A610031)资助项目~~

摘　　要：探讨肝细胞生长因子(HGF)基因转染人淋巴瘤细胞系Raji细胞后,拮抗足叶乙甙(VP-16)诱导细胞凋亡的研究。将三种细胞:未转染Raji细胞、空载体pVITRO2-mcs转染细胞和HGF基因转染细胞,分成正常对照组和经VP-16处理的药物组。采用Western blot法验证HGF蛋白的表达;CCK-8法检测诱导Raji细胞凋亡的药物浓度;通过透射电镜、流式细胞术、吖啶橙(AO)染色、苏木精–伊红(HE)染色等方法观察Raji细胞的凋亡情况,并进行相关分析。结果显示:Western blot法验证了HGF蛋白质的表达;CCK-8法显示100μg/mL足叶乙甙可明显抑制Raji细胞增殖;透射电镜下可发现典型的凋亡细胞;流式检测结果表明:给药组与正常组相比,三组细胞的凋亡率明显升高(P<0.01),提示VP-16具有诱导细胞凋亡的作用;但给药组间:HGF基因转染组凋亡率明显低于未转染组(P<0.05)和空载体pVITRO2-mcs转染组(P<0.05),提示HGF基因转染可明显抑制VP-16诱导的Raji细胞的凋亡,AO染色和HE染色结果也同样提示HGF具有拮抗VP-16诱导的细胞凋亡效应。To investigate hepatocyte growth factor （HGF） gene transfection and its inhibitory effect on the apoptosis of human lymphoma cell line Raji cells induced by VP-16, three cell lines （non-transfected Raji cells, empty vector transfected Raji cells and pVITRO2-mcs-HGF gene transfected Raji cells） were designed into normal groups and drug groups. Western blot was used to estimate the HGF protein level. CCK-8 assay was used to mea- sure proliferated inhibition on Raji cells induced by VP-16. Quantitative and qualitative analyses on the apoptosis of Raji cells were performed through transmission electron microscopy, flow cytometry, acridine orange （AO） fluorescent staining and HE staining. The results showed that HGF protein expression was evaluated. CCK-8 assay revealed that VP-16 can inhibit the proliferation of Raji significantly at the concentration of 100 μg/mL. Typical morpho- logic changes of cell apoptosis were observed under transmission electronic microscope. Flow cytometry results showed that the apoptotic rates of drug groups were significantly higher than the control groups （P〈0.01）, indicated that the apoptotic rate was enhanced after treated with VP-16. The apoptotic rates of HGF gene transfection groups were significantly lower than the non-transfected group （P〈0.05） and empty vector transfected group （P〈0.05）, in- dicated that HGF gene transfection can significantly protect Raji cells from apoptosis induced by VP-16. The results of acridine orange （AO） fluorescent staining and HE staining indicated that HGF can decrease the apoptotic rate of Raji cells induced by VP-16.

关 键 词：肝细胞生长因子 基因转染 凋亡 

分 类 号：R733.1[医药卫生—肿瘤]