siRNA介导的MAPKp42沉默诱导HeLa细胞凋亡

目的应用siRNA沉默HeLa细胞MAPKp42，研究其对细胞存活的影响。方法体外合成靶向MAPKp42的siRNA-1和siRNA-2，并用脂质体转染HeLa细胞，Western印迹法和免疫组织化学法检测p42^MPAK的表达；电镜观察，TUNEL法测定细胞凋亡，Annexin-PI法测定不同凋亡时期细胞的分布。结果siRNA-1和siRNA-2均可使HeLa细胞MAPK p42沉默，使p42^MAPK表达下调，与阴性对照组相比分别降低2、5倍和3．2倍。电镜观察证实，siRNA-1和siRNA-2转染组部分细胞丧失表面微绒毛，细胞内空泡增加，细胞核染色质边集。处理组HeLa细胞早期凋亡率显著增加，其中siRNA-1组晚期凋亡率也明显增加。结论siRNA介导的MAPKp42沉默可以诱导HeLa细胞凋亡。

Small interfering RNA-mediated MA PK p42 silencing induces apoptosis of HeLa cells

Objective To observe the effect of small interfering RNA (siRNA)-induced MA PK p42 silencing on the survival of HeLa cells. Methods Two siRNAs targeting at the MA PK p42 gene and one random siRNA were synthesized respectively by Silencer?siRNA Construction Kit and transfected into HeLa cells by Lipofectamin?2000. The expression of p42MAPK in the transfected HeLa cells was analyzed by Western blotting and immunohistochemistry, and the morphology of cells were observed with electron microscope. TUNEL assay and Annsxin V/PI staining were employed for detecting the cell apoptosis. Results The expression of p42MAPK in the HeLa cells was remarkably suppressed after transfection with the two siRNAs, reduced by about 2.5 and 3.2 folds respectively in comparison with the negative control. Chromatin margination in the cell nuclei were observed in the transfected cells, and TUNEL assay and Annexin V/PI staining further confirmed the occurrence of cell apoptosis. Conclusion In vitro MAPK p42 siRNA-1 and siRNA-2 transfection can specifically silence the gene expression and induce apoptosis of HeLa cells.