| 摘要: |
| 目的 探讨鞘氨醇激酶2(SphK2)在5-氟尿嘧啶(5-FU)诱导的人结肠癌HCT116细胞凋亡中的作用。方法 采用Hoechst33342染色检测5-FU作用48 h后HCT116细胞凋亡形态学的变化；流式细胞术检测5-FU作用48 h后HCT116细胞的凋亡率,并分析SphK2的干扰或过表达对5-FU诱导的HCT116细胞凋亡率的影响；用蛋白质印迹法观察5-FU诱导HCT116细胞SphK2活性的改变及干扰或过表达SphK2对5-FU诱导凋亡中凋亡标记蛋白的影响。结果 5-FU能诱导HCT116细胞凋亡,细胞出现了明显的核固缩、染色质凝集等凋亡形态变化。细胞SphK2被干扰后,与未被干扰的细胞相比,5-FU诱导后的细胞凋亡率增高(P<0.01),凋亡标记蛋白表达水平升高；而过表达SphK2的细胞在5-FU作用下其凋亡率较对照组(空质粒处理)下降(P<0.01),凋亡标记蛋白表达水平亦下降；5-FU可诱导磷酸化SphK2水平增高。结论 SphK2对5-FU诱导的HCT116细胞凋亡具有负调控作用。 |
| 关键词: 5-氟尿嘧啶 HCT116细胞 鞘氨醇激酶2 细胞凋亡 |
| DOI：10.3724/SP.J.1008.2014.01060 |
| 投稿时间：2014-02-13修订日期：2014-05-26 |
| 基金项目:湛江市科技攻关计划(2013B01124)，广东医学院博士启动课题 (B2012032). |
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| Sphingosine kinase 2 negatively regulates 5-fluorouracil-induced apoptosis in human colon cancer cells |
| XIAO Min*,WU Xian-jin,CAI Mu-fa,YI Wei-lian |
(Department of Clinical Diagnosis, Affiliated Hospital of Guangdong Medical College, Zhanjiang 524001, Guangdong, China
*Corresponding authors) |
| Abstract: |
| Objective To explore the role of sphingosine kinase 2 (SphK2) in 5-fluorouracil (5-FU)-induced apoptosis in human colon cancer HCT116 cells. Methods Hoechst33342 staining was used to examine the apoptosis morphological changes of HCT116 cells 48 h after treatment with 5-FU; flow cytometry was used to detect the apoptotic ratio of HCT116 cells and the effects of SphK2 (down-regulation and over-expression) on 5-FU-induced apoptosis were analyzed. Western blotting analysis was used to examine the changes in the activated forms of SphK2 and apoptotic marker protein in HCT116 cells after exposure to 5-FU. Results 5-FU induced significant apoptosis in HCT116 cells as reflected by apoptosis characteristics-condensation and fragmentation of chromatin. Interference of SphK2 significantly increased 5-FU-induced apoptosis in HCT116 cells compared with non-interfered group (P<0.01), accompanied by increased apoptotic marker protein. On the contrary, the apoptosis ratio of the cells with SphK2 over-expression was significantly decreased compared with vehicle plasmid-transfected group (P<0.01), accompanied by decreased apoptotic marker protein. Moreover, 5-FU greatly increased activated SphK2. Conclusion SphK2 negatively regulates 5-FU-induced apoptosis in HCT116 cells. |
| Key words: 5-fluorouracil HCT116 cell sphingosine kinase 2 apoptosis |
