Effects of G250 promoter controlled conditionally replicative adenovirus expressing Ki67-siRNA on renal cancer cell
Effects of G250 promoter controlled conditionally replicative adenovirus expressing Ki67-siRNA on renal cancer cell
复制标题
G250启动子控制的条件复制腺病毒表达Ki67-siRNA对肾癌细胞的影响
DOI：
10.1111/j.1349-7006.2012.02380.x
复制
发表时间：
2012-10-01
期刊：
影响因子：
5.7
通讯作者：
Zheng, Junnian
中科院分区：
文献类型：
--
作者：
Liu, Junjie;Fang, Lin;Zheng, Junnian
复制型腺病毒（RCAd）已广泛应用于肿瘤基因治疗，肿瘤选择性是使用复制型腺病毒的关键。本实验研究了溶瘤病毒的抗肿瘤特性，溶瘤病毒的E1 A基因受肾细胞癌特异性启动子G250启动子的控制。构建的溶瘤病毒G250-Ki 67携带Ki 67-siRNA转基因，G250-ZD 55-Ki 67也缺失E1 B-55 KD。Western blot和免疫组化染色检测E1 A和Ki 67的表达，结晶紫染色和细胞活力测定检测E1 A和Ki 67的细胞毒活性。G250-Ki 67和G250-ZD 55-Ki 67腺病毒能在786-O和OSRC细胞中表达E1 A蛋白，而在ACHN和HK-2细胞中不表达。腺病毒感染786-O和OSRC细胞后，Ki 67基因的表达受到抑制。G250-ZD 55-Ki 67和G250-Ki 67对786-O细胞的杀伤作用明显高于OSRC细胞。各组腺病毒均能抑制786-O细胞和OSRC细胞的增殖。但G250-ZD 55-Ki 67和G250-Ki 67对786-O细胞的作用强于对OSRC细胞的作用。G250-ZD 55-Ki 67对肾癌细胞的抗肿瘤活性明显高于G250-Ki 67。G250启动子来源的CRAds携带Ki 67-siRNA，可在表达G250抗原的肾癌细胞中高效扩增和表达Ki 67-siRNA，抑制肾癌细胞增殖并诱导凋亡。这些结果表明，表达Ki 67-siRNA的G250特异性溶瘤腺病毒适用于人肾透明细胞癌的治疗。
Replication-competent adenovirus (RCAd) has been used extensively in cancer gene therapy, and tumor-selection is critical for the use of replication-competent adenovirus. Here we investigated the anti-tumor characterization of oncolytic virus, whose E1A gene is under the control of a renal cell carcinoma specific promoter the G250 promoter. The constructed oncolytic virus G250-Ki67 is armed with transgene of Ki67-siRNA, and G250-ZD55-Ki67 also with E1B-55 KD deleted. The tumor-specific expression of E1A and Ki67 was demonstrated by Western blot and immunohistochemistry staining, and the tumor-specific cytotoxicity was assessed by crystal violet staining and cell viability assays. The G250-Ki67 and G250-ZD55-Ki67 adenoviruses could express E1A protein in 786-O and OSRC cell lines but not in ACHN and HK-2 cell lines. The expression of Ki67 gene in 786-O and OSRC cell lines were suppressed by these adenoviruses. The cytotoxic effects induced by G250-ZD55-Ki67 and G250-Ki67 were more obvious on the 786-O cell lines than on the OSRC cell lines. Each group of adenoviruses could inhibit the proliferation of the 786-O cells and OSRC cells. However, the effects induced by G250-ZD55-Ki67 and G250-Ki67 on 786-O cells were stronger than on OSRC cells. Moreover, G250-ZD55-Ki67 had enhanced antitumor activities in these renal cancer cells compared with G250-Ki67. G250 promoter-derived CRAds carrying Ki67-siRNA could highly amplify and express Ki67-siRNA in renal cancer cells with expression of G250 antigen, inhibit renal cancer cells proliferation and induce apoptosis. These results demonstrated that the G250-specific oncolytic adenovirus expressing Ki67-siRNA is applicable for human renal clear cell cancer therapy.