目的 探讨4′-甲醚-黄芩素(4-MS)对人绒毛膜癌JAR细胞的增殖抑制作用及诱导凋亡的相关机制。方法 应用MTT法、激光共聚焦扫描显微镜、流式细胞术和实时定量PCR法,体外观察4-MS对JAR细胞的影响。结果 不同质量浓度(10、20、40mg/L)的4-MS对JAR细胞均有增殖抑制作用,呈剂量依赖,作用72h后其增殖抑制率分别为(14.14±0.75)%、(34.34±2.99)%、(61.11±2.99)%(P<0.01);4-MS作用72h后细胞内Ca²⁺浓度分别为81.3±6.7、103.3±5.9、120.7±11.0,与对照组相比差异有显著性(P<0.05)。40mg/L4-MS作用12、24、36h后其早期凋亡率分别为(2.36±0.19)%、(4.22±2.44)%、(7.34±0.56)%,明显高于对照组(P<0.01)。20、40mg/L4-MS作用48h后hTERT mRNA的表达量为0.05±0.01和0.02±0.01,明显低于对照组(P<0.01)。结论 4-MS能抑制人绒毛膜癌JAR细胞的增殖,诱导凋亡,其机制与提高细胞内Ca²⁺浓度、降低hTERT mRNA的表达有关。

Objective To investigate the inhibition and the mechanism of apoptosis inducement to the cell proliferation by 4'-methyl ether-scutellarein(4-MS)on human choriocarcinoma JAR cell line.Methods The techniques of MTT assay,flow cytometry,laser scanning confocal microscopy(LSCM),and real-time quantitative PCR were used to study the effect of 4-MS on JAR cells in vitro.Results 4-MS had does-and time-dependent inhibitory effects on proliferation of JAR cells.After the JAR cells were treated by 4-MS in different concentrations(10,20,and 40 mg/L)for 72 h,the inhibitory rates were(14.14±0.75)%,(34.34±2.99)%,and(61.11±2.99)%(P<0.01),and the intracellular Ca²⁺ concentrations were 81.3±6.7,103.3±5.9,and 120.7±11.0,with the significant difference compared to the control group(P<0.05),respectively.After the JAR cells were treated by 4-MS(40 mg/L)for 12,24,and 36 h,the early apoptosis rates were(2.36±0.19)%,(4.22±2.44)%,and(7.34±0.56)%,much higher than that in the control group(P<0.01),respectively.After the JAR cells were treated by 4-MS(20 and 40 mg/L)for 48 h,the expressions of hTERT mRNA were 0.05±0.01 and 0.02±0.01,much lower than that in the control group(P<0.01).Conclusion The proliferation can be inhibited and the apoptosis of human choriocarcinoma JAR cell line can be induced by 4-MS.Its mechanism is related to the increase of intracellular Ca2+ concentration and the decrease of hTERT mRNA expression.

江苏省自然科学基金资助项目(BK2003104)