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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
by
李潮 刘晓溪 付海燕 杜红阳
in
半胱氨酸天冬氨酸蛋白酶3
/ 细胞凋亡
/ 雷公藤内酯
/ 黑色素瘤
2015
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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
by
李潮 刘晓溪 付海燕 杜红阳
in
半胱氨酸天冬氨酸蛋白酶3
/ 细胞凋亡
/ 雷公藤内酯
/ 黑色素瘤
2015
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Journal Article
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
2015
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Overview
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。
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