摘要
目的观察黑色素瘤分化相关基因7(MDA-7/IL-24)基因对人肝癌细胞Hep3B和正常的肝细胞L02的作用,并且探讨其该作用机制。方法将携带人MDA-7/IL-24基因的腺病毒Ad.mda-7感染人正常肝细胞L02和肝癌细胞HeP3B,逆转录-聚合酶链式反应(RT-PCR)和ELISA方法观察MDA-7/IL-24基因的表达,噻唑蓝染色法(MTT)观察MDA-7/IL-24对肝癌细胞的生长抑制,Hoechst染色和Annexin-V和PI双染后流式细胞仪检测二种细胞的凋亡,利用PI染色后流式细胞仪检测细胞周期,RT-PCR方法检测bcl-2的表达变化。结果Ad.mda-7能介导外源基因MDA-7/IL-24在肝癌细胞株Hep3B和正常细胞L02中高效表达,细胞培养上清液中MDA-7/IL-24蛋白的表达(Hep3B:L02分别为790:810ng/L)。MDA-7/IL-24能明显抑制肝癌细胞的生长(抑制率分别是83%和1.2%),能促进肝癌细胞的凋亡(58%:2.2%),阻滞肝癌细胞在G_2/M期(48.29%:7.95%)。而对正常的肝细胞没有促凋亡和增殖阻滞作用;能明显的抑制Hep3B的凋亡抑制基因bcl-2的表达。结论Ad.mda-7能介导MDA-7/IL-24基因在人肝癌细胞中高效表达,选择性的杀伤肝癌细胞Hep3B,促进细胞增殖阻滞,其机制是通过抑制bcl-2的表达诱导肿瘤细胞凋亡。
Objective To investigate the effect of MDA/IL-24 on the human hepatocullular carcinoma line Hep3B in vitro. Methods The MDA-7/IL-24 gene was transfected into human hepatocullular carcinoma cell line Hep3B and normal liver cell line L02 with an replication-incompetent adenovirus vector. The expression of MDA7/IL-24 and bcl-2 in Hep3B and L02 cells was detected by RT-PCR and ELISA assay. MTT assay and flow cytometry were used to study tumor cell proliferation and cell cycle. Hoechst staining and Annexin-V and PI staining were performed to study MDA-7/IL-24 gene expressed in Hep3B and L02 cells. Results The protein concentration of MDA-7 in supernatant of Hep3B and L02 cells was 790 and 810 pg/mL respectively. MDA-7/IL-24 could significantly inhibit the human hepatocellular carcinoma cell Hep3B growth （inhibition rate was 83% and 1.2% ） , promote the apoptosis （58% and 2.2% ） and block the cells in GJM in vitro （48.29% ,7.95% ） ,but couldnt promote the apoptosis and inhibit the proliferation of L02 cells. The gene expression of bcl-2 was significantly deceased in Hep3B but not in L02 cells. Conclusion MDA-7/IL-24 selectively induces growth suppression and apoptosls of hepatocellular carcinoma lines Hep3B in vitro but not normal liver cell L02 by inhibiting the expression of bcl-2.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2007年第7期792-794,共3页
Chinese Journal of Experimental Surgery
基金
湖北省科技攻关重点项目(2006AA304B05)
