氯化锂对卵巢癌细胞增殖凋亡影响及其机制探讨  

Effect of lithium chloride on proliferation and apoptosis of ovarian cancer cell

作　　者：廖菁[1] 丁冬[1] 杨宗元[1] 胥琴[1] 翁丹卉[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,湖北武汉430030

出　　处：《中华肿瘤防治杂志》2014年第4期241-245,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81000979;81272859);中央高校基本科研业务费资助(2012TS058);湖北省自然科学基金(2011CDB542)

摘　　要：目的:观察氯化锂(lithium chloride,LiCl)对卵巢癌SKOV3细胞增殖活性及凋亡的影响,并探讨其作用机制。方法:使用20(A组)、40(B组)和60(C组)mmol/L LiCl处理SKOV3细胞,正常对照组用等体积的细胞培养基培养。采用CCK8法观察LiCl对卵巢癌SKOV3细胞增殖活性的影响;流式细胞仪检测LiCl作用后细胞周期变化及凋亡情况。蛋白质印迹法检测细胞糖原合成酶激酶-3β(glycogen synthase kinase3β,GSK-3β)、磷酸化糖原合成酶激酶-3β(phosphorglation-glycogen synthase kinase3β,p-GSK-3β)及有丝分裂阻滞缺陷蛋白2(mitotic arrest deficient2,MAD2)蛋白表达的变化。结果:不同浓度LiCl分别作用于SKOV3细胞48h后,A组细胞增殖活性为(80.64±1.06)%,与B组(57.18±1.56)%比较,差异有统计学意义,P=0.001;与C组(43.18±6.12)%比较,差异有统计学意义,P<0.001。B组作用24h细胞增殖活性为(75.12±9.35)%,48h为(57.18±1.56)%,72h为(35.36±1.00)%,与24h组相比,48h组(P=0.158)和72h组(P=0.036)细胞增殖活性降低。LiCl能有效抑制SKOV3细胞增殖,呈时间与剂量依赖性。不同浓度LiCl作用96h后SKOV3细胞的凋亡率相对于对照组升高,A组为(7.82±0.87)%,P=0.037;B组为(21.09±1.57)%,P<0.001;C组为(27.83±0.47)%,P<0.001,差异均有统计学意义。不同浓度LiCl作用48h后,细胞出现了明显的G2/M期阻滞,G2/M期的比例由对照组的(10.23±1.50)%依次升高,A组为(19.9±4.16)%,P=0.352;B组为(30.29±0.51)%,P=0.045;C组为(39.32±1.11)%,P=0.009,差异有统计学意义。蛋白质印迹法检测结果显示,LiCl浓度升高,p-GSK-3β蛋白相对表达量升高,A组为(54.39±2.75)%,P=0.019;B组为(72.39±2.89)%,P=0.009;C组为(86.57±2.52)%,P=0.005,差异有统计学意义。LiCl浓度升高,MAD2蛋白相对表达量升高,A组为(52.29±1.34)%,P=0.041;B组为(58.35±1.36)%,P=0.030;C组为(72.07±2.93)%,P=0.006,差异有统计学意义。结论:LiCl能有效抑制卵巢癌SKOV3细胞的增殖,并诱导其凋亡;上调MAD2增强有丝分裂检测点功能,�OBJECTIVE： To investigate the influence of LiC1 on proliferation and apoptosis in human ovarian cancer cell and the possible mechanism. METHODS： SKOV3 cells were treated with 20 mmol/L（A group） ,40 mmol/L（B group） and 60 mmol/L（C group） LiC1 separately. The growth inhibitory effects were evaluated by CCK8 assay. Cell cycle and ap optosis were observed by flow cytometry. Western blot was used to detect the expressions of proteins GSK-3β, p-GSK-3β and MAD2. RESULTS： The proliferation of SKOV3 cells was inhibited by LiC1. The proliferative activities were decreased to （80.64±1.06）% for A group, （57.18±1.56）%for B group, （43.18±6.12）% for C group respectively after treated with LiC1 for 48 h （compared to the A group,P：0. 001 for B group and P=0. 001 for C group）. The proliferative activi- ties were decreased to （75.12±9.35）0//00 for 24 h group,（57.18±1.56）O//oo for 48 h group,（35.36±1.00）% and for 72 h group after treated with B group （compared to the 24 h group,P values were 0. 158 for 48 h group and 0. 036 for 72 h group）. LiC1 could inhibit the proliferation of SKOV3 cells in a time-and dose-dependent manner. Treatment of LiC1 for 96 h induced apoptosis of SKOV3 cells and the apoptosis rates were increased to （7. 82 ± 0.87）0//oo for A group,（21. 09±1.57）% for B group, （27. 83±0.47）~//00 for C group compared with the control group （P= 0. 037 for A group, P〈0. 001 for B group and C group）. After treated with the different concentrations above for 48 h,the percents of G2/M phase were increased from （10.23±1.50）% of the control to （19. 9±4. 16）% for A group,（30.29±0. 51）%0 for B group, （39.32 ~ 1.11）% for C group （P values were 0. 352 for A group,0. 045 for B group and 0. 009 for C group）. Flow cytometry revealed that the SKOV3 cells were arrested in G2/M phase. LiC1 treatment for 48 h could also up-regulate the expression level of p-GSK-313 as well as MAD2 while the expression of total GSK-3~t and was not change. T

关 键 词：卵巢肿瘤 氯化锂 增殖 凋亡 糖原合成酶激酶-3Β 有丝分裂阻滞缺陷蛋白2 

分 类 号：R737.31[医药卫生—肿瘤]