氯化钴诱导HIF-1α增高拮抗过氧化氢引起的人骨肉瘤MG-63细胞凋亡

Antagonism effects of cobalt chloride with increasing HIF-1α on apoptosis of human osteosarcoma MG-63 cells induced by H_2O_2

[目的]验证氯化钴抑制过氧化氢诱导的人骨肉瘤MG-63细胞的凋亡作用并探讨其可能的分子机制.[方法]将人骨肉瘤MG-63细胞分为正常对照组、过氧化氢组、过氧化氢+氯化钴预处理组、过氧化氢+氯化钴+YC-1组.应用MTT法测定细胞存活率;流式细胞术测定细胞凋亡;免疫荧光染色、蛋白印迹法检测HIF-1α表达.[结果]与过氧化氢组比较,氯化钴预处理组细胞存活率明显增加(P<0.05);凋亡细胞比率降低.缺氧可激活HIF-1α的表达并诱导其转位,100μmol/L氯化钴可明显增加HIF-1α蛋白的表达(P<0.01).[结论]氯化钴可抑制过氧化氢诱导的人骨肉瘤MG-63细胞的凋亡,其分子机制可能与激活HIF-1α的表达有关.

OBJECTIVE To study the antagonism effects and its possible molecular mechanism of cobalt chloride（CoC12） on apoptosis of human osteosarcoma MG-63 cells induced by H202. METHODS MG-63 cells were divided into 4 groups of control group, H202 group, H202 ＋ COC12 and H202 ＋ COC12 ＋ YC-1 group, and the survival rate of MG-63 cells was measured by MTT, the apoptosis of MG-63 cells was detected by flow cytometry, and the expression of HIF-la was determined by immunofluorescence and Western Blot. RESULTS As compared with H202 group, the survival rate of MG-63 cells increased significantly in pretreatment group with COC12 （P 〈0.05） and apoptosis rate of MG-63 cells decreased. Hypoxia could activate the expression of HIF-la and induced its translocation. 100 μmol/L of COC12 were significantly up-regulated the expression of HIF-la protein in MG-63 cells. CONCLUSION The COC12 inhibits the apoptosis of human osteosarcoma MG-63 cells induced by H202, and its molecular mechanism is related to activating the expression of HIF-la.