IL-27基因修饰Eca109细胞的体外体内凋亡作用的研究

目的 研究IL-27基因转染人食管癌Eca109细胞株后在体外和体内对细胞凋亡的影响,从而探讨其抗肿瘤作用及其机制.方法 基因转染的方法 建立稳定表达IL-27基因的人食管癌细胞株(Eca109/IL-27),观察Eca109/IL-27细胞生长情况、移植瘤的生长情况及生存期,用流式细胞技术检测体外和体内的细胞凋亡率和Fas的表达,采用Western blot检测Survivin 的表达.结果 成功建立稳定转染的Eca109/IL-27细胞株,RT-PCR示Eca109/IL-27细胞中有mIL-27 p28和EBI3亚基基因表达,而Eca109/LXSN和Eca109细胞中未见表达(P＜0.01),IL-27基因转染不影响人食管癌细胞株的体外生长和细胞凋亡,Fas和Survivin的表达无变化(P＞0.05);但体内Eca109/IL-27移植瘤生长较Eca109和Eca109/LXSN滞后,生存时间延长(P＜0.05),肿瘤组织细胞凋亡率增高(P＜0.05),细胞表面Fas的表达增加(P＜0.05),Survivin表达降低(P＜0.05).结论 IL-27在体外不影响细胞凋亡,体内可能通过降低Survivin的表达,上调Fas的表达,诱导细胞凋亡产生抗肿瘤作用.

In vitro and in vivo apoptosis of IL-27 gene-transduced Eca109 cells

Objective To investigate the effects of interleukin-27 g ene transduction on cell apoptosis of esophageal carcinoma cells (Eca109) in v itro and in vivo, as well as the potential mechanism involved in. Methods The retrovirus vector was used to transduce the IL-27 gene into human Eca109. Stable clones expre ssing IL-27 (Eca109/IL-27) were obtained by selecting with G418. The expressio n of IL-27p28 and IL-27EBI3 genes in Eca109/IL-27 was detected by RT-PCR. MTT colo rimetry was used to assess the in vitro growth of Eca109/IL-27. The growth of t r ansplanted tumor in nude mice was also observed. Flow cytometry was used to ana lyze the cell apoptosis and the expression of Fas in vitro and in vivo. The expr ession of survivin was observed by Western blotting. Results Ec a109/IL-27 cells were set up successfully. The expression of IL-27p28 and EBI3 in Eca109/IL-27 was positive, contrasted to Eca109/LXSN and Eca109 cells (P<0.01). The grow th r ate and cell apoptosis of Eca109/IL-27 were similar to those of parental cell a n d neo-vector transfected cells. The in vitro expression of Fas and survivin wer e also the same (P>0.05). Growth of Eca109/IL-27 tumor was significantly slowed down posttransplantation in nude mice (P<0.05). Cell apoptosis ra te of tissue transplanted by Eca109/IL-27 cells significantly increased. The ex pression of Fas was upregulated but surviving expression was significantly decre ased (P<0.05). Conclusion IL-27 has no effects on cell apo ptosis in vitro. But in vivo, IL-27 can enhance antitumor immunity by decreasing survivin expression, upregu lating Fas expression, and inducing cell apoptosis.