过表达α1,3-岩藻糖基转移酶Ⅶ通过增强p38MAPK信号通路抑制UVC照射诱导的人肝癌SMMC-7721细胞的凋亡

Overexpression of α1,3-fucosyltransferaseⅦ attenuates UVC-induced apoptosis of SMMC-7721 cells through enhancing p38MAPK-signaling pathway

目的初步探讨α1,3-岩藻糖基转移酶Ⅶ(简称FUT7)在UVC照射诱导的人肝癌SMMC-7721(简称7721)细胞凋亡过程中的作用及机制。方法用RT-PCR检测转染pcDNA3-FUT7质粒的7721细胞及转染空质粒pcDNA3的7721细胞中FUT7基因的转录水平;用流式细胞仪检测细胞表面FUT7产物SLex的表达水平;利用DAPI染核计算UVC照射后细胞的凋亡比率;用结晶紫染色法测定细胞的存活率;利用Western blot检测Caspase3的剪切情况及p38MAPK、JNK1/2和ERK1/2的磷酸化水平。结果过表达FUT7能抑制UVC照射诱导的7721细胞凋亡,同时还增强了细胞中p38MAPK信号通路的活性,而用p38MAPK的特异性抑制剂处理则可削弱FUT7的抗凋亡作用。结论在UVC照射诱导的7721细胞凋亡过程中FUT7具有抗凋亡的作用,这种作用可能部分通过增强p38MAPK信号通路活性而实现。

Objective To study the role of α1 ,3-fucosyltransferase Ⅶ （FUT7）in UVC-induced apoptosis of human hepatocarcinoma cell line SMMC-7721 （7721）. Methods The expression of FUT7 in both pcDNA3-FUT7 transfectant and mock pcDNA3 transfectant cells were detected by RT- PCR. The level of SLe^x was detected by flow cytometry. Apoptotic percentage was detected by DAPI staining. Survival percentage was detected by crystal violet staining. The levels of cleaved caspase3 and phosphorylation of p38MAPK, JNK1/2 and ERK1/2 were detected by Western blot. Results Overexpression of FUT7 attenuated UVC-induced apoptosis of 7721 and enhanced p38MAPK-signaling pathway. In addition,pretreatment with specific p38MAPK inhibitor reduced the anti-apoptotic effect of FUT7. Conclusions FUT7 functions as an anti-apoptotic molecule during UVC-induced apoptosis,at least in part,through enhancing p38MAPK-signaling pathway.