人质膜型唾液酸酶对丁酸钠诱导细胞凋亡的影响及其机制  

Effect of hmSD on apoptosis induced by NaBT and its mechanism

作　　者：禹彬[1] 许莉[1] 孙连坤[1] 李晓洁[1] 李扬[1] 赵雪俭[1] 

机构地区：[1]吉林大学基础医学院病理生理学教研室

出　　处：《吉林大学学报（医学版）》2008年第1期49-52,F0002,共5页Journal of Jilin University：Medicine Edition

基　　金：教育部留学基金资助课题(2003);吉林省科技厅基金资助课题(20070728-1)

摘　　要：目的:研究人质膜型唾液酸酶(hmSD)对丁酸钠(NaBT)诱导人雄激素非依赖型前列腺癌细胞(PC3细胞)凋亡的影响及其可能机制。方法:以前列腺癌细胞PC3和稳定转染hmSD的前列腺癌细胞(PC3-hmSD)为靶细胞,分别设对照组、NaBT2.5mmol·L-1组、NaBT5.0mmol·L-1组、NaBT10.0mmol·L-1组,采用MTT法检测细胞存活率,吖啶橙/溴化乙啶(AO/EB)染色方法检测细胞凋亡情况,DCFH-DA激光共聚焦法检测细胞内ROS,Western blotting方法检测Bcl-XL、P65和Caspase9蛋白表达。结果:①NaBT作用48和72h时,PC3-hmSD细胞的生存率明显高于PC3细胞(P<0.05);②NaBT5.0mmol·L-1作用4h,PC3-hmSD组细胞凋亡数明显少于PC3细胞(P<0.05);③NaBT诱导细胞内活性氧产生,PC3-hmSD细胞与PC3细胞比较差异无显著性;④NaBT作用下,与PC3细胞比较,PC3-hmSD细胞Bcl-XL、P65蛋白表达水平增强,而Caspase9蛋白表达水平减低(P<0.05)。结论:①hmSD具有抵抗NaBT诱导细胞凋亡的作用;②hmSD对NaBT诱导活性氧产生无抑制作用;③hmSD抗凋亡作用可能与Bcl-XL和P65表达上调、Caspase9表达下调有关。Objective To study the effect of hmSD on apoptosis of PC3 ceils induced by NaBT and its mechanism. Methods PC3 cells and PC3-hmSD stablely transfected with hmSD were used as target cells. Four groups were set up： control, NaBT 2.5 mmol·L^-1, NaBT 5.0 mmol·L^-1 and NaBT 10.0 mmol·L^-1. The survival rate of cells was detected by MTT assay, apoptotic rate was determined with AO/EB assay, intracellular ROS was examined with DCFH-DA staining, the expressions of Bcl-XL, P65 and Caspase 9 were analyzed by Western blotting method. Results ①The survival rates of PC3-hmSD cells treated with NaBT for 48 and 72h were higher than that of PC3 cells （P〈0.05）. ② The apoptotic rate of PC3-hmSD cells treated with NaBT 5.0 mmol· L^-1 for 4 h was lower than that of PC3 cells （P〈0.05）. ③ There was no difference of ROS between both cell lines treated with NaBT.④ The result of Western blotting showed that the expression of Bcl-XL decreased in NaBT-treated PC3 cells, but it increased in PC3-hmSD cells. Compared with control group, the expressions of P65 and Caspase 9 increased in NaBT-treated cells, but the expression of P65 increased and the expression of Caspase 9 decreased in hmSD overexpressed cells （P〈0.05）. Conclusion ① NaBT can induce the apoptosis of PC3 cells, and hmSD can reverse it. ②The pathway of hmSD against apoptosis may be no relation with the suppressed ROS synthesis. ③ The antiapoptotic mechanism of hmSD may be involved in down-regulation of Caspase 9 and up-regulation of P65 and Bcl-XL expressions.

关 键 词：人质膜型唾液酸酶 前列腺癌细胞 丁酸钠 细胞凋亡 

分 类 号：Q255[生物学—细胞生物学]