人绒毛膜促性腺激素和胸苷合成酶在氟尿嘧啶核苷连续诱导耐药绒癌细胞系构建过程中的动态表达  

The role of β-human chorionic gonadotropin secretion and expression level of thymidylate synthase gene in fluorodeoxyuridine-resistance prediction of choriocarcinoma

作　　者：韩冰[1,2] 向阳[1] 王铮[3,4] 王云[3,4] 张浩[1] 黄尚志[3,4] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院妇产科,北京100730 [2]苏州大学附属第一医院妇产科 [3]中国医学科学院基础医学研究所医学遗传学系 [4]北京协和医学院基础学院

出　　处：《现代妇产科进展》2011年第11期886-889,共4页Progress in Obstetrics and Gynecology

基　　金：国家自然基金(No:30973178)

摘　　要：目的:通过检测氟尿嘧啶脱氧核苷(fluorodeoxyuridine,FUDR)连续诱导的耐药绒癌细胞构建过程中β-人绒毛膜促性腺激素(β-human chorionic gonadotropin,β-HCG)和胸苷合成酶(thymidylate synthase,TS)的动态表达,检验β-HCG分泌水平和TS基因的表达量是否适用于筛查绒癌细胞对该药耐药的指标。方法:采用连续诱导法,利用FUDR诱导人绒癌细胞系JeG-3,建立耐药细胞株JeG-3/FUDRC2;用化学发光法检测培养液上清中激素分泌水平,用荧光定量PCR检测TS mRNA在诱导过程中的动态表达。结果:(1)历时12月成功建立了绒癌耐药细胞系JeG-3/FUDRC2,其耐药指数为12.46±2.22;(2)随着药物浓度和肿瘤细胞耐药性的逐渐增加,绒癌细胞的β-HCG分泌最初表现为上调,诱导药物达到一定浓度时其分泌达到高峰,而后骤然下降,并有随药物浓度和耐药性的继续增加而逐渐下降的趋势;(3)低浓度药物诱导后,TS基因表达明显上调,但随着药物浓度和耐药性增加,其表达不升反降,甚至低于亲本细胞TS基因的表达量。结论:成功建立了连续诱导耐药的绒癌细胞株JeG-3/FUDRC2。绒癌细胞β-HCG分泌和TSmRNA表达与FUDR作用浓度以及耐药性无明显相关性。就目前的研究结果而言,β-HCG分泌和TS mRNA的表达水平不适宜用作绒癌细胞是否对FUDR耐药的指标。Objective：To assess dynamic profiling of β-human chorionic gonadotropin（β-HCG）and thymidylate synthase（TS）transcript level in fluorodeoxyuridine（FUDR）-resistant JeG-3 sub-lines by using consecutive feeding method.Methods：Consecutive feeding methods were used to establish FUDR-resistance JeG-3 sub-line.The resistant index（RI）was measured by cell counting kit-8（CCK-8）assay.The chemo-luminescence assay was used to detect the hormone secretion.qRT-PCR was used to detect the mRNA expression level of thymidylate synthase（TS）in the sub-lines during the different concentration of FUDR-exposure.Results：The RI of JeG-3/FUDRC2 were 12.46±2.22.When exposed to low concentration of FUDR,the upward β-HCG secretion and mRNA level of TS gene were detected.However,upon increase of FUDR concentration,both of them decreased sharply.Conclusions：We established the FUDR-resistant sub-line of JeG-3 by using consecutive inducing method.The β-HCG secretion and TS mRNA expression level is not in accordance with the concentration of FUDR-exposure and the increasing RI.The data suggests that β-HCG secretion and TS mRNA expression level may not be used as a biomarker to predict the chemo-resistant in FUDR-based chemotherapy

关 键 词：绒癌 耐药 氟尿嘧啶核苷 胸苷合成酶 绒毛膜促性腺激素 

分 类 号：R737.33[医药卫生—肿瘤]