马鞭草C部位对人绒癌JAR细胞hCG分泌的影响和作用机制

Effect of Verbena officinals C on Inhibiting Secretion of hCG and Intriguring Apoptosis of JAR Cell

目的研究马鞭草C部位对绒毛膜癌JAR细胞绒毛膜促性腺激素(hCG)分泌的影响及其作用机制。方法不同浓度马鞭草C部位(马鞭草C部位终浓度10,2040mg/ml)作用于体外培养的JAR细胞,分别于第12,24,48,72h取上清液,采用βhCG单克隆抗体酶免定量法检测hCG含量;琼脂糖凝胶电泳观察用药后细胞DNA断裂情况;流式细胞仪检测细胞周期分布及细胞凋亡率;透射电镜观察细胞超微结构的变化。结果马鞭草C部位对绒毛膜癌JAR细胞分泌hCG产生明显的抑制作用,呈时间和剂量相关性。在该药物作用下,细胞DNA琼脂糖凝胶电泳出现“阶梯状条带”;流式细胞仪显示细胞周期分布改变,G2/M期细胞比例增高,并出现不同程度的凋亡峰;超微结构显示典型的凋亡形态学改变。结论一定浓度的马鞭草C部位对JAR细胞hCG分泌有明显的抑制作用,其作用机制是诱导细胞凋亡。

AIM:To study the inhibition effect of Verbena officinals C on secretion of hCG and the mechanism of the human chorionic carcinoma cell line JAR in vitro.METHOD:The human chorionic carcinoma cell line JAR was used for research.The suspension for each well containing 10,20,40 mg/ml Verbena officinals C was added to 96 well microtitre culture plate,cultured for 12,24,48 h and 72 h.The supernatants were collected for detecting hCGβ-subunit.DNA agarose gel electrophoresis was used to observe the evidence of apoptosis;Flow cytometry was used for the cell cycle distribution and estimating apoptosis;the morphology of apoptosis cells was observed by transmission electron microscopy.RESULT:Verbena officinals C significantly inhibited the secretion of hCGβ-subunit by JAR cells in vitro,and showed the correlationship between concentration and time.Verbena officinals C increased the proportion of cells in the G_~2 /M phase and induced apoptosis of JAR cells.“Ladder strait” was observed by agarose gel electrophoresis,and the morphological changes of apoptosis appeared following being treated with Verbena officinals C. CONCLUSION: Verbena officinals C can inhibit the secretion of hCGβ-subunit by JAR cells,and induce apoptosis of JAR cells.