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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 study the effect of biological agents 1α, 25-dihydroxyvitamin D3 on JAR of choriocarcinoma cell line and to explore its mechanism. Methods: Calcitriol at different concentrations (10-8,10-7,10-6 mol / L) was detected by MTT assay at different time points (1 day, 3 days and 6 days) On the proliferation of JAR cells; the effect of calcitriol on the cell cycle of JAR was detected by flow cytometry; the expression of vitamin D receptor (VDR) mRNA was detected by RT-PCR; the expression of VDR protein was detected by Western blot. Results: MTT assay showed that Jurum cells treated with 10-6mol / L calcitriol for 3 days had an inhibitory rate of (22.74 ± 12.08)% (P <0.05), (29.64 ± 8.66)% (P <0.01), (49.74 ± 17.42)%, respectively (P <0.01), showing a significant time-dose effect. Compared with the control group, the percentage of cells in G1 phase of calcitriol group increased from 39.78% to 59.81% (P <0.01), and the percentage of cells in S phase decreased from 54.6% to 33.08% (P <0.01). Calcitriol upregulates VDR mRNA and protein expression in JAR cells. CONCLUSION: Calcitriol can significantly inhibit JAR proliferation and induce JAR differentiation in choriocarcinoma cell lines. Its mechanism may be related to the up-regulation of VDR mRNA by calcitriol and the increase of VDR protein expression.