摘要
目的:研究生物制剂1α,25-二羟基维生素D3(calcitriol)对绒毛膜癌细胞株JAR的影响,探讨其作用机制。方法:采用四甲基偶氮唑蓝(MTT)比色法检测不同浓度(10-8、10-7、10-6mol/L)calcitriol在不同的时间点(1天、3天、6天)对JAR细胞增殖的影响;流式细胞仪检测calcitriol对JAR细胞周期的影响;RT-PCR检测维生素D受体(VDR)mRNA水平的表达;Western blot检测VDR蛋白表达。结果:MTT比色法检测10-6mol/Lcalcitriol作用JAR细胞3天,10-7、10-6mol/L calcitriol作用6天抑制率分别为(22.74±12.08)%(P<0.05),(29.64±8.66)%(P<0.01),(49.74±17.42)%(P<0.01),呈明显的时间-剂量效应关系。calcitriol作用JAR细胞的半数抑制浓度(IC50)组与对照组比较,细胞周期G1期细胞比例从39.78%增至59.81%(P<0.01),S期细胞数百分比从54.6%减至33.08%(P<0.01)。calcitriol上调JAR细胞的VDR mRNA及蛋白表达水平。结论:calcitriol能明显抑制绒毛膜癌细胞株JAR增殖,并诱导JAR分化,其作用机制与calcitriol上调VDR mRNA,进而增加VDR蛋白的表达有关。
Objective：To observe the effects of 1α,25-Dihydroxyvitamin D3 on JAR cells,and to evaluate its mechanisms.Methods：The human choriocarcinoma cells（JAR）cultured in vitro were treated by 1α,25-Dihydroxyvitamin D3 with concentrations 10-8,10-7,10-6mol/L for periods of 1d,3d,6d.Cell proliferation was evaluated by MTT assay.Distribution of cell cycle was determined by flow cytometry.The expression of VDR mRNA and protein was respectively examined by RT-PCR assay and Western blot assay.Results：The results of MTT showed that calcitriol could inhibit the proliferative activity of JAR cells significantly,inhabiting growth in a dose-dependent and time-dependent manner.These phenomena were observed after calcitriol of 10-6mol/L treatment for 3d（P0.05）,10-6 or 10-7mol/L for 6d（P0.01）.Distribution of cell cycle was determined by flow cytometry.After cultured with calcitriol of 10-7mol/L for 6d（IC50 group）,the percentage of cells in G1 phase was elevated from 39.78% to 59.81%,and the percentage of cells in S phase was obviously decreased from 54.6% to 33.08%,comparing with those of control group.Calcitriol may up-regulate expression of VDR mRNA and protein.Conclusion：Calcitriol can both inhibit proliferation and induce differentiation of human choriocarcinoma cell line JAR by up-regulating expression of VDR mRNA and protein.
出处
《现代妇产科进展》
CSCD
北大核心
2011年第8期614-617,共4页
Progress in Obstetrics and Gynecology
基金
广东省卫生厅"五个一"科教兴医工程重点课题资助(No:9603)
