论文部分内容阅读
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 the effect of hepatocyte growth factor (HGF) gene on the apoptosis of human lymphoma cell line Raji induced by etoposide (VP-16). The three kinds of cells were transfected into the untransfected Raji cells, empty vector pVITRO2-mcs transfected cells and HGF gene and divided into normal control group and VP-16 treated group. The expression of HGF protein was detected by Western blot. The concentration of drug induced apoptosis of Raji cells was detected by CCK-8. The apoptosis of Raji cells was detected by transmission electron microscopy, flow cytometry, acridine orange (AO) staining, hematoxylin-eosin (HE) Staining and other methods to observe the apoptosis of Raji cells and make correlation analysis. The results showed that the expression of HGF protein was verified by Western blot. The CCK-8 assay showed that 100μg / mL etoposide could inhibit the proliferation of Raji cells. The apoptotic cells were found by transmission electron microscopy. The results of flow cytometry showed that Compared with the normal group, the apoptotic rate of the three groups of cells was significantly increased (P <0.01), suggesting that VP-16 can induce apoptosis; however, the apoptosis rate of the HGF gene transfected group (P <0.05) and pVITRO2-mcs transfected group (P <0.05), which indicated that HGF gene transfection could obviously inhibit the apoptosis of Raji cells induced by VP-16. AO staining and HE The staining results also suggest that HGF antagonizes VP-16-induced apoptosis.