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目的探讨黑色素瘤分化相关基因-7/白介素-24基因(mda-7/IL-24)对不同种类肝癌细胞的选择性杀伤作用。方法将携带人 mda-7/IL-24基因的复制缺陷型腺病毒(Ad.mda-7)感染人正常肝细胞和各种肝癌细胞,通过逆转录聚合酶链反应(RT-PCR)和酶联免疫吸附(ELISA)实验观察mda-7/IL-24基因的表达,MTT 法观察肝癌细胞的生长抑制,Hoechst 染色及 Annexin-V 和 PI 双染后流式细胞仪检测细胞的凋亡情况及用流式细胞仪检测细胞周期。结果 RT-PCR 和 ELISA 提示Ad.mda-7能介导外源基因 mda-7/IL-24在各种肝癌细胞株和正常肝细胞中高效表达。MTT 实验结果提示 mda-7/IL-24能明显抑制各种肝癌细胞的生长,Hoechst 染色和流式细胞仪检测提示 mda-7/IL-24能选择性杀伤肝癌细胞,细胞周期分析提示 mda-7/IL-24阻滞肝癌细胞在 G2/M 期,同时对正常的肝细胞没有促凋亡和增殖阻滞作用。结论 mda-7/IL-24基因能选择性杀伤各种肝癌细胞,促进细胞增殖阻滞及诱导细胞凋亡。
Objective To investigate the selective killing effect of mda-7 / IL-24 gene on different kinds of hepatocellular carcinoma cells. Methods Ad-mda-7 cells harboring mda-7 / IL-24 gene were infected with human hepatocellular carcinoma cells and normal liver cells by reverse transcription polymerase chain reaction (RT-PCR) and enzyme The expression of mda-7 / IL-24 gene was detected by ELISA. The growth inhibition of HCC cells was observed by MTT assay. The apoptosis of HCC cells was detected by flow cytometry with Hoechst staining and Annexin-V and PI double staining. Cell cycle was detected by flow cytometry. Results RT-PCR and ELISA suggested that Ad.mda-7 could efficiently express mda-7 / IL-24 in various hepatocellular carcinoma cell lines and normal liver cells. MTT assay indicated that mda-7 / IL-24 could significantly inhibit the growth of various hepatocarcinoma cells. Hoechst staining and flow cytometry suggested that mda-7 / IL-24 could selectively kill hepatoma cells. Cell cycle analysis suggested that mda- 7 / IL-24 block hepatoma cells in G2 / M phase, while no normal apoptosis on the liver cells and proliferation arrest. Conclusion The mda-7 / IL-24 gene can selectively kill various hepatocarcinoma cells, promote cell proliferation arrest and induce apoptosis.