间充质干细胞介导的TRAIL可控性表达及其杀伤人肝癌细胞的活性研究

目的 研究肿瘤坏死因子相关凋亡诱导配体(TRAIL)在间充质干细胞中可控性表达及其对肿瘤细胞的杀伤活性,探索间充质干细胞用于肿瘤治疗的前景.方法 构建TRAIL可控性表达的腺病毒载体Ad-Tet-TRE-TRAIL,包装获得重组腺病毒后,感染小鼠间充质干细胞,免疫印迹法和酶联免疫吸附法测定间充质干细胞TRAIL可控性表达和分泌.四甲基偶氮唑盐(MTT)法测定TRAIL抑制肝癌细胞生长的能力.膜联蛋白(Annexin)-V/碘化丙啶(PI)染色和流式细胞术测定TRAIL对肝癌细胞的杀伤活性.结果 重组腺病毒Ad-Tet-TRE-TRAIL感染的小鼠间充质干细胞能够在强力霉素的控制下表达和分泌TRAIL,并显著抑制SMMC-7402人肝癌细胞生长,其机制是诱导肝癌细胞凋亡,结果显示SMMC-7402细胞在杀伤24、48 h后的存活率分别为66.5%±4.8%和42.9%±6.5%,而对照组(未加强力霉素)的细胞存活率分别为97.3%±2.2%和99.4%±4.7%.结论 间充质干细胞介导TRAIL可控性表达进而有效诱导肝癌细胞凋亡并抑制肝癌细胞生长,为肿瘤的细胞治疗提供了新的策略.

Abstract：

Objective To study the controllabe expression of soluble tumor necrosis factor-related apoptosis-iuducing ligand (TRAIL) in mesenchymal stem cells and evaluate its potential tumoricidal effects in cancer therapy. Methods The controllable TRAIL expression vector of Ad-Tet-TRE-TRAIL was established in an adenovirus vector for transfection into murine mesenchymal stem cells. The controllable expression and secretion of TRAIL were detected by Western blot and enzyme-linked immunosorbent assay.The viability of hepatocellular carcinoma cells was determined by MTT assay. The tumoricidal activity of TRAIL was determined by Annexin-V/PI staining and flow cytometry. Results The murine expression model of TRAIL was successfully established in the presence of doxycycline. The secreted TRAIL in cell culture medium could efficaciously suppress the growth of human hepatocellular carcinoma SMMC-7402 by induced apoptosis. The cell viability of SMMC-7402 was 66. 5% ± 4. 8% and 42.9% ± 6. 5% at post-treatment versus 97.3% ±2. 2% and 99. 4% ±4. 7% in the control group at 24 h and 48 h. Conclusion The controllable TRAIL expression mediated by mesenchymal stem cells kills human hepatocellular carcinoma cells effectively. And it may provide a novel therapeutic strategy for hepatocellular carcinoma.

Killing effects of controllable expression of tumor necrosis factor-related apoptosis-inducing ligand from mesenchymal stem cells on hepatocellular carcinoma cells

Objective To study the controllabe expression of soluble tumor necrosis factor-related apoptosis-iuducing ligand (TRAIL) in mesenchymal stem cells and evaluate its potential tumoricidal effects in cancer therapy. Methods The controllable TRAIL expression vector of Ad-Tet-TRE-TRAIL was established in an adenovirus vector for transfection into murine mesenchymal stem cells. The controllable expression and secretion of TRAIL were detected by Western blot and enzyme-linked immunosorbent assay.The viability of hepatocellular carcinoma cells was determined by MTT assay. The tumoricidal activity of TRAIL was determined by Annexin-V/PI staining and flow cytometry. Results The murine expression model of TRAIL was successfully established in the presence of doxycycline. The secreted TRAIL in cell culture medium could efficaciously suppress the growth of human hepatocellular carcinoma SMMC-7402 by induced apoptosis. The cell viability of SMMC-7402 was 66. 5% ± 4. 8% and 42.9% ± 6. 5% at post-treatment versus 97.3% ±2. 2% and 99. 4% ±4. 7% in the control group at 24 h and 48 h. Conclusion The controllable TRAIL expression mediated by mesenchymal stem cells kills human hepatocellular carcinoma cells effectively. And it may provide a novel therapeutic strategy for hepatocellular carcinoma.