雷公藤内酯醇诱导人脉络膜黑色素瘤株OCM-1凋亡的机制研究  

Mechanism of apoptosis of triptolide induced human choroidal melanoma cell line OCM-1

作　　者：李潮[1] 刘晓溪[2] 付海燕[3] 杜红阳[4] 

机构地区：[1]辽宁锦州辽宁医学院附属第一医院病理科,121001 [2]辽宁锦州辽宁医学院附属第一医院儿科,121001 [3]辽宁锦州辽宁医学院附属第一医院重症医学科,121001 [4]辽宁锦州辽宁医学院附属第一医院皮肤科,121001

出　　处：《解放军医学杂志》2015年第2期117-120,共4页Medical Journal of Chinese People's Liberation Army

基　　金：辽宁省教育厅科学技术研究项目(L2010275);辽宁省自然科学基金(2013022059);辽宁医学院青年科技启动基金(Y2011Z010)~~

摘　　要：目的探讨雷公藤内酯醇(TPL)对人脉络膜黑色素瘤株OCM-1增殖的影响,分析其诱导人OCM-1细胞凋亡的作用机制。方法将OCM-1细胞分为对照组和实验组,实验组加入20μl不同终浓度(5、10、20、40、80、160nmol/L)的TPL,对照组加入等量无血清RPMI 1640培养液,分别作用24、48、72h。采用MTT法检测TPL对OCM-1细胞增殖的影响,瑞氏-吉姆萨染色观察细胞形态学变化,流式细胞术检测不同浓度TPL诱导后OCM-1细胞的凋亡比例,Western blotting检测TPL诱导后凋亡相关蛋白Bcl-2、Bax、survivin及caspase-3的表达。结果 TPL可抑制OCM-1细胞增殖,48h及72h的IC50值分别为56.14±6.72、15.57±4.28nmol/L。TPL诱导OCM-1细胞后,在瑞氏-吉姆萨染色中可观察到凋亡小体。流式细胞术在不同浓度TPL诱导后的OCM-1细胞中均检测到亚二倍体凋亡峰。TPL诱导OCM-1细胞凋亡过程中,Western blotting检测显示Bax蛋白表达上调(P<0.01),Bcl-2、survivin蛋白表达下调(P<0.01),并可检测到活化的caspase-3。结论 TPL可抑制人OCM-1的增殖并诱导其凋亡,Bax、Bcl-2、survivin及caspase-3蛋白在其中可能具有重要作用。Objective To investigate the effect oftriptolide （TPL） on proliferation of human choroidal melanoma cell line OCM-1, and analyze the mechanism of apoptosis induction. Methods OCM-1 cells were divided into control and experimental groups. 201μl TPL （final concentration was 5, 10, 20, 40, 80 and 160 nmol/L respectively） was used for experimental group, and an equal volume of serum-free RPMI 1640 medium for the control group. The culture time was 24, 48 and 72 hours, respectively. The effect of TPL on OCM-1 cell proliferation was detected by MTT assay. The morphological change of apoptotic cells was observed by Switzerland-Giemsa staining. The expressions of TPL-induced OCM-1 apoptosis-associated proteins Bcl-2, Bax, survivin and caspase-3 were determined by Western blotting. Results TPL suppressed OCM-1 cell proliferation, at 48 and 72h, and ICs0 concentration was 56.14±6.72 and 15.57 ± 4.28 nmol/L respectively. After OCM-1 cells were cultured with TPL, apoptotic bodies were observed with Switzerland-Giemsa staining and subdiploid apoptotic peaks by flow cytometry. The expression of protein Bax was up-regulated （P〈0.01）, while Bcl-2 and survivin down-regulated （P〈0.01 ）, and caspase-3 was activated during apoptosis induced by TPL. Conclusion TPL could inhibit proliferation and induce apoptosis of human choroidal melanoma cell line OCM-1, and the proteins survivin, Bcl-2, Bax and caspase-3 play an important role in the induction of OCM-1 apoptosis.

关 键 词：雷公藤内酯 黑色素瘤 细胞凋亡 半胱氨酸天冬氨酸蛋白酶3 

分 类 号：R739.7[医药卫生—肿瘤]