Department of General Surgery, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China.
Hepatobiliary Pancreat Dis Int. 2010 Dec;9(6):615-21.
Melanoma differentiation-associated gene-7 (MDA-7)/interleukin-24 (IL-24) is a novel tumor suppressor gene, which has suppressor activity in a broad spectrum of human cancer cells. We investigated the effect of the replication-competent oncolytic adenovirus SG600-IL24 and replication-incompetent adenovirus Ad.IL-24, both expressing human MDA-7/IL-24 on the hepatocellular carcinoma cell lines HepG2, Hep3B, SMMC-7721, HCCLM3, and the normal liver cell line L02.
Hepatocellular carcinoma cell lines and the normal liver cell line were infected with SG600-IL24 and Ad.IL-24. The mRNA and protein expression of MDA-7/IL-24 in infected cells was confirmed by RT-PCR, ELISA, and Western blotting. MTT assay was used to investigate the proliferation effect. Hoechst staining and Annexin-V and PI staining were performed to study the MDA-7/IL-24 gene expressed in HCC cell lines and the normal liver cell line. Flow cytometry was used to analyse the cell cycle.
RT-PCR, ELISA and Western blotting confirmed that the exogenous MDA-7/IL-24 gene was highly expressed in cells infected with SG600-IL24. MTT and apoptosis detection indicated that SG600-IL24 induced growth suppression, promoted apoptosis, and blocked cancer cell lines in the G2/M phase in hepatocellular carcinoma cell lines but not in the normal liver cell line.
SG600-IL24 selectively induces growth suppression and apoptosis in hepatocellular carcinoma cell lines in vitro but not in the normal liver cell line L02. Compared with Ad.IL-24, SG600-IL24 dramatically enhances antitumor activity in hepatocellular carcinoma cell lines.
黑色素瘤分化相关基因-7（MDA-7）/白细胞介素-24（IL-24）是一种新型肿瘤抑制基因，对广泛的人类癌细胞具有抑制活性。我们研究了复制型溶瘤腺病毒 SG600-IL24 和复制缺陷型腺病毒 Ad.IL-24 对肝癌细胞系 HepG2、Hep3B、SMMC-7721、HCCLM3 和正常肝细胞系 L02 的影响，这两种腺病毒都表达了人 MDA-7/IL-24。
用 SG600-IL24 和 Ad.IL-24 感染肝癌细胞系和正常肝细胞系。通过 RT-PCR、ELISA 和 Western blot 验证感染细胞中 MDA-7/IL-24 的 mRNA 和蛋白表达。MTT 法检测增殖效果。Hoechst 染色、Annexin-V 和 PI 染色研究 MDA-7/IL-24 基因在 HCC 细胞系和正常肝细胞系中的表达。流式细胞术分析细胞周期。
RT-PCR、ELISA 和 Western blot 证实 SG600-IL24 感染的细胞中外源 MDA-7/IL-24 基因高表达。MTT 和凋亡检测表明，SG600-IL24 诱导生长抑制、促进凋亡，并阻断肝癌细胞系在 G2/M 期，但不阻断正常肝细胞系。
SG600-IL24 体外选择性诱导肝癌细胞系生长抑制和凋亡，但不诱导正常肝细胞系 L02。与 Ad.IL-24 相比，SG600-IL24 显著增强了肝癌细胞系的抗肿瘤活性。
