Objective:To observe the effect of the artesunate(ART) on cellular proliferation in vitro,to search for the possible anti-tumor mechanism of ART on endometrial carcinoma at the molecular level and to provide the exper...Objective:To observe the effect of the artesunate(ART) on cellular proliferation in vitro,to search for the possible anti-tumor mechanism of ART on endometrial carcinoma at the molecular level and to provide the experimental and theoretical foundations for the clinical applications of ART.Methods:The cell proliferation was observed by microscope;MTT was used to examine the effects of ART on proliferation of HEC-1B cells,and flow cytometric analysis was used to detect cell cycle and apoptosis.The human endometrial carcinoma HEC-1B cells were conventionally cultured;ART was administered with a concentration of 40 μg/ml before the total RNA were extracted.mRNA expression of Survivin,Caspase-3,N-Cadherin,E-Cadherin,Fibronectin1 and Cox-2 were detected using RT-PCR.Results:ART reduced proliferation in human endometrial carcinoma cell line HEC-1B in a dose-and time-dependent effect.The cells of G0/G1 stage were significantly increased(P<0.05),but the cells of G2/M stages were significantly decreased(P<0.05),so it has shown that the cell cycle was probably blocked in G0/G1 stage.After intervention with ART at 20 and 80 μg/ml for 48 h,cellular apoptosis rate respectively was(36.42±0.77)% and(11.77±0.58)%,and the difference was statistically significant compared with the control([6.64±0.19]%,P<0.01).The expression of Cox-2 mRNA in the ART group was lower than those of control group,yet the expression of Caspase-3 and E-Cadherin mRNA in the ART group was higher than those of control group.Conclusion:ART can inhibit HEC-1B cell growth and proliferation in a dose-and time-dependent manner.Furthermore,ART can induce apoptosis in a dose-dependent manner.ART is able to downregulate Cox-2 mRNA expression and to upregulate E-Cadherin and Caspase-3 mRNA expression.So we can conclude that ART could induce the endometrial carcinoma HEC-1B cell apoptosis and inhibit tumor cell proliferation.展开更多
目的:探讨在子宫内膜癌细胞中重组人程序性细胞死亡蛋白5(recombinant human programmed cell death protein 5,rhPDCD5)对紫杉醇化疗的促进作用。方法:子宫内膜癌KLE细胞培养完成后,通过重组人rh PDCD5(20μg/ml)处理KLE细胞,再分别以0...目的:探讨在子宫内膜癌细胞中重组人程序性细胞死亡蛋白5(recombinant human programmed cell death protein 5,rhPDCD5)对紫杉醇化疗的促进作用。方法:子宫内膜癌KLE细胞培养完成后,通过重组人rh PDCD5(20μg/ml)处理KLE细胞,再分别以0、1.0、5.0、10.0、50μmol/L紫杉醇(paclitaxel,PTX)处理24 h或以10μmol/L PTX处理0、12、24、48 h,提取细胞总RNA及蛋白后,CCK法检测KLE细胞的增殖情况,流式细胞术检测KLE细胞凋亡情况,实时定量PCR检测KLE细胞中PDCD5mRNA的表达量,实时定量PCR或Western blotting测定凋亡相关基因的Bax、Bcl2、caspase-3 mRNA或蛋白水平的变化。结果:PTX对PDCD5表达的促进作用具有剂量依赖性和时间依赖性;PTX的最佳作用浓度为10μmol/L,最佳作用时间为24 h。rh PDCD5明显增强紫杉醇对KLE细胞的抑制作用。CCK实验、流式细胞术及Western blotting检测显示:PTX+rhPDCD5联合处理组KLE细胞的增殖抑制率和凋亡率均较PTX组明显增加、pro-caspase 3的表达量明显增加(均P<0.01)。促进凋亡蛋白Bax和抑制凋亡蛋白Bcl2的比值亦较明显增加(P<0.01)。结论:rhPDCD5可协同PTX抑制子宫内膜癌KLE细胞的增殖、促进细胞的凋亡,可明显增强KLE细胞对PTX的药物敏感性。展开更多
文摘Objective:To observe the effect of the artesunate(ART) on cellular proliferation in vitro,to search for the possible anti-tumor mechanism of ART on endometrial carcinoma at the molecular level and to provide the experimental and theoretical foundations for the clinical applications of ART.Methods:The cell proliferation was observed by microscope;MTT was used to examine the effects of ART on proliferation of HEC-1B cells,and flow cytometric analysis was used to detect cell cycle and apoptosis.The human endometrial carcinoma HEC-1B cells were conventionally cultured;ART was administered with a concentration of 40 μg/ml before the total RNA were extracted.mRNA expression of Survivin,Caspase-3,N-Cadherin,E-Cadherin,Fibronectin1 and Cox-2 were detected using RT-PCR.Results:ART reduced proliferation in human endometrial carcinoma cell line HEC-1B in a dose-and time-dependent effect.The cells of G0/G1 stage were significantly increased(P<0.05),but the cells of G2/M stages were significantly decreased(P<0.05),so it has shown that the cell cycle was probably blocked in G0/G1 stage.After intervention with ART at 20 and 80 μg/ml for 48 h,cellular apoptosis rate respectively was(36.42±0.77)% and(11.77±0.58)%,and the difference was statistically significant compared with the control([6.64±0.19]%,P<0.01).The expression of Cox-2 mRNA in the ART group was lower than those of control group,yet the expression of Caspase-3 and E-Cadherin mRNA in the ART group was higher than those of control group.Conclusion:ART can inhibit HEC-1B cell growth and proliferation in a dose-and time-dependent manner.Furthermore,ART can induce apoptosis in a dose-dependent manner.ART is able to downregulate Cox-2 mRNA expression and to upregulate E-Cadherin and Caspase-3 mRNA expression.So we can conclude that ART could induce the endometrial carcinoma HEC-1B cell apoptosis and inhibit tumor cell proliferation.
