2-甲氧雌二醇对子宫颈癌细胞及其移植瘤的作用

目的探讨2-甲氧雌二醇（2-ME2）对HeLaS3细胞的作用,以及对鼠荷人宫颈癌移植瘤的作用。方法应用细胞增殖检测和细胞周期分析,检测加入不同浓度2-ME2的HeLaS3细胞及二甲基亚砜（实验组）的生长情况和细胞周期变化,以仅加入二甲基亚砜的HeLaS3细胞作为对照（对照组）;采用荧光显微镜观察HeLaS3细胞的形态变化,以DNA电泳检测细胞凋亡情况;采用蛋白印迹法检测2-ME,对诱生型一氧化氮合酶（iNOS）的表达。此外,建立鼠荷人宫颈癌移植瘤模型,口饲2-ME2（75mg／kg）14d,检测肿瘤体积的变化,并通过原位细胞凋亡检测,分析肿瘤中的细胞凋亡情况。结果1μmol／L2-ME2作用48h后,HeLaS3新生细胞数是对照组的81％（P〈0．05）,作用96h,是对照组的19％（P〈0．01）。5μmol／L2-ME2作用20h后,凋亡细胞由对照组的4％增至16％,G2／M期细胞由对照组的16％增至55％（P〈0．01）;荧光显微镜发现典型的核固缩及形态异常的有丝分裂中期细胞,DNA电泳发现典型的梯状DNA条带。iNOS的表达随2-ME：作用时间延长和浓度增加而变化,与凋亡变化相似,而且iNOS抑制剂1400W抑制了2-ME2诱导的细胞凋亡。在动物实验中,与对照组相比,2-ME2治疗组宫颈癌移植瘤体积下降34％;原位细胞检测发现,2-ME2治疗组肿瘤切片中凋亡和坏死细胞增多。结论2-ME2具有抑制人类宫颈癌细胞HeLaS3及鼠荷人宫颈癌移植瘤生长的作用,有可能成为宫颈癌抑制剂,

Effect of 2-methoxyestradiol on human cervical cancer cells HeLaS3 and xenografts

OBJECTIVE: To evaluate the effects of 2-methoxyestradiol (2-ME(2)) on human cervical cancer HeLaS3 cells and cervical cancer xenografts. METHODS: Cell proliferation assay and cell cycle analysis were used to measure HeLaS3 cell growth and cell cycle progression after 2-ME(2) treatment. Fluorescent microscopy to observe the cell morphology and DNA electrophoresis to measure apoptosis. In addition, the effect of 2-ME(2) on the expression of inducible nitric oxide synthase (iNOS) was measured by Western blot. Moreover, human cervical cancer model was set up using HeLaS3 cells and 2-ME(2) (75 mg/kg) was orally given for 14 d. Tumor volume was determined and apoptosis was detected by in situ cell death. RESULTS: Newly-formed cell amount in treated group was 81% of that in control group after 1 micromol/L 2-ME(2) treatment for 48 h (P < 0.05), and was 19% of that in control group after 2-ME(2) treatment for 96 h (P < 0.01). G(2)/M phase cells were increased to 55% from 16% of the control (P < 0.01), and apoptotic cells were increased to 16% from 4% of the control, after 5 micromol/L 2-ME(2) treatment for 20 h. Nuclear condensation and abnormal metaphase cells were found by fluorescent microscopy. Typical DNA ladder was found by DNA electrophoresis. And the expression of iNOS was increased by 2-ME(2) in a time- and concentration-dependent manner, in parallel with apoptosis. Moreover, apoptosis was prevented by the iNOS inhibitor 1400W. In vivo, tumor volume was reduced 34% while compared with the control group. In situ cell death detection found more apoptotic and necrotic cells in 2-ME(2)-treated group. CONCLUSIONS: 2-ME(2) inhibits human cervical cancer HeLaS3 cells and tumor growth in cervical cancer xenografts. Thus 2-ME(2) has the therapeutic potential for cervical carcinoma.