洋椿苦素对人宫颈癌Hela细胞增殖的抑制作用研究  

The study of cedrelone inhibiting human cervix cancer Hela cells proliferation

作　　者：刘怡[1] 陈辉[2] 王凤[1] 阙昌田 杨胜群[1] 曾南[1] 

机构地区：[1]成都中医药大学药学院中药材标准化教育部重点实验室四川省中药资源系统研究与开发利用重点实验室--省部共建国家重点实验室培育基地,成都611137 [2]电子科技大学附属医院.四川省人民医院,成都610072

出　　处：《中药药理与临床》2016年第1期65-69,共5页Pharmacology and Clinics of Chinese Materia Medica

摘　　要：目的:研究洋椿苦素(cedrelone)体外对人宫颈癌Hela细胞增殖的抑制作用,并探讨其诱导细胞凋亡的作用机制。方法:采用CCK-8法计算不同浓度的洋椿苦素作用人宫颈癌Hela细胞后的IC50及增殖抑制率;Hoechst 33258荧光染色法和流式细胞术Annexin V-FITC/PI双染法检测洋椿苦素对Hela细胞凋亡的诱导作用;流式细胞术PI单染法检测洋椿苦素对Hela细胞生长周期的影响;Real-time PCR法测定CDK2、cyclinA mRNA表达。结果:不同浓度的洋椿苦素作用Hela细胞48小时后均能抑制其生长,其IC50值为1.4μg/ml。洋椿苦素作用24小时后,Hela细胞呈典型的凋亡形态学改变,细胞体积缩小,胞核致密浓染,呈半月状或呈碎块状。0.21μg/ml、0.42μg/ml、1.05μg/ml、1.47μg/ml、1.89μg/ml、4.22μg/ml的洋椿苦素作用Hela细胞24小时后的凋亡率分别为3%、4.1%、6.2%、7%、8.4%、58.5%。洋椿苦素作用于Hela细胞24小时后,可使细胞生长停滞在S期,即DNA合成期,并剂量依赖性地下调CDK2、cyclinA mRNA表达水平。结论:洋椿苦素对人宫颈癌Hela细胞增殖具有良好抑制作用,作用发挥与下调CDK2、cyclin A mRNA表达从而抑制Hela细胞S期DNA合成,诱导其凋亡有关。Objective： To investgate the effect of cedrelone on proliferation of human cervix cancer Hela cells in vitro and to explore the molecular mechanism. Methods： The CCK-8 assay was employed to dertermine the IC50 values and inhibiton rate of cedrelone on Hela cells. Cells apoptosis were detected by Hoechst 33258 fluorescent staining and Annexin V-FITC / PI double staining assay via flow cytometric analysis. Cell cycle was detected by PI single staining through flow cytometric analysis. The expression of CDK2 and cyclinA was dectected by Real-time PCR.Results： After the treatment with different concentration of cedrelone for 48 hours,Hela cells growth was inhibited and the IC50 of cedrelone on Hela cells was 3. 345 μg / ml. Marked morphological changes of cell apoptosis such as condensation of chromatin and nuclear fragmentations were found after Hela cells were treated with cedrelone for 24 hours. After treated with 0. 5,1,2. 5,3. 5,4. 5,10μg / ml cedrelone for 24 hours,the inhibition rate of Hela cells was 3%,4. 1%,6. 2%,7%,8. 4%,58. 5%,respectively and the control group was 3. 4%. The cell cycle was arrested in S phase via the treatment with cedrelone for 24 hours,which lead to inhibit DNA synthesis. The expression of CDK2,cyclinA mRNA was down-regulated. Conclusion： It was indicated that cedrelone could influence the proliferation of Hela cells and induce apoptosis via suppressing DNA synthesis in S phase via down-regulating the expression of CDK2 and cyclinA.

关 键 词：洋椿苦素 HELA细胞 细胞凋亡 细胞周期 细胞周期调控因子 流式细胞术 

分 类 号：R285[医药卫生—中药学]