凝血酶敏感蛋白-1对人肝癌细胞株HCCLM3凋亡的作用

目的 研究凝血酶敏感蛋白-1(TSP-1)对人肝癌细胞株HCCLM3凋亡的体外诱导作用及机制.方法 采用流式细胞术检测TSP-1及其受体CD36、CD47诱导HCCLM3的细胞凋亡率,应用电镜分析作用后的形态变化,逆转录聚合酶链反应(RT-PCR)分析作用后HCCLM3细胞Caspase-3 mRNA表达的变化.结果 TSP-1组凋亡率[(12.44±0.72)%]显著高于对照组[(4.31±0.29)%]和CD47阻断组[(4.99±0.12)%],P<0.01.CD36阻断组[(9.99±0.57)%]高于对照组或CD47阻断组,低于TSP-1组(P<0.01).电镜观察:对照组和C1D47阻断组细胞生长旺盛.TSP-1组和CD36阻断组细胞凋亡率增加,细胞呈现各种凋亡的表现.TSP-1组Caspase-3 mRNA的表达TSP-1组(0.652±0.024)和CD36阻断组(0.615±0.020)显著高于对照组(0.398±0.033)和CD47阻断组(0.432±0.019),P<0.01.结论 TSP-1可诱导人肝癌细胞株HCCLM3的凋亡,TSP-1与受体CD47结合后上调Caspase-3的表达可能是作用途径之一.Abstract：Objective To study the effect and mechanism of thrombospondin- 1 on the apoptosis of human hepatocarcinoma cell HCCLM3. Methods Detecting the role of TSP-1, CD36, CD47 on apoptosis rate of HCCLM3 by the flow cytometry and the change of cell ultractructure by transmission electron microscope,then detecting the influence of TSP-1, CD36, CD47 on the expression of Caspase-3 in HCCLM3. Results The apoptosis rate was respectively significantly higher in TSP-1 group [(12. 44 ±0. 72)%] than that in control group [(4. 31 ±0. 29)%] or CD47-blocked group[(4. 99 ±0. 12)%],P < 0. 01. The apoptosis rate was was respectively significantly higher in CD36 -blocked group [(9. 99 ±0. 57)%]than that in control group and CD47 -blocked group,was lower than in the TSP-1 group(P <0. 01). The morphological changes were: grow vigorously and many divided nuleus in control group and CD47- blocked group. Apoptosis cell increased in TSP- 1 group and CD36 -blocked group: less cellular module in cytoplasm, many nucleus pieces and concentrate. The expression of Caspase-3 were higher in TSP-1 group (0. 652 ± 0. 024) and CD36-blocked group (0. 615 ± 0. 020) than that in control group (0. 398 ±0. 033)and CD47-blocked group (0. 432 ± 0. 019) (P < 0. 01). Conclusions TSP-1 can induce apoptosis of HCCLM3 cell line through combined with CD47 which can increase the expression Caspase -3.

Apoptosis of human hepatic carcinoma cell line HCCLM3 induced by thrombospondin -1

Hepatocellur carcinoma Thrombospondin-1 HCCLM3 Apoptosis Caspase-3