摘要
目的观察Pou class 6同源框2(Pou6f2)基因对泌乳素腺瘤MMQ细胞增殖和分泌能力的影响。方法敲低Pou6f2实验:将细胞分为对照组和实验1、2组,按Lipofectamine~? 3000转染试剂盒说明书分别转染control-siRNA、Pou6f2-siRNA-1、Pou6f2-siRNA-2,72 h后收集细胞。过表达Pou6f2实验:将细胞分为对照组和实验组,按Lipofectamine~? 3000转染试剂盒说明书分别转染空载质粒、野生型Pou6f2质粒,72 h后收集细胞。采用Western blotting法检测细胞中Pou6f2表达量,MTS法测定细胞增殖活性(A_(490)值),ELISA法测定细胞泌乳素(PRL)分泌水平。结果敲低Pou6f2实验中,与对照组比较,实验1、2组MMQ细胞Pou6f2表达量降低、增殖的A_(490)值增加、分泌PRL水平升高(P均<0. 01),实验1、2组各指标差异均无统计学意义。过表达Pou6f2实验中,与对照组比较,实验组MMQ细胞Pou6f2表达量增加、增殖的A_(490)值降低、分泌PRL水平下降(P均<0. 01)。结论 Pou6f2可以抑制泌乳素腺瘤MMQ细胞的增殖及PRL分泌能力,有望成为泌乳素腺瘤治疗的新靶点。
Objective To observe the effects of Pou class 6 homeobox 2 (Pou6f2) on the proliferation and secretion of prolactin of rat prolactinoma MMQ cells. Methods The experiment of knocking down Pou6f2:The MMQ cells were divided into the control group and the experimental groups 1 and 2,and the control-siRNA,Pou6f2-siRNA-1,Pou6f2-siRNA-2 were transfected into cells respectively according to the instructions of Lipofectamine 3000 transfection kit,and the cells were collected 72 hours later.The experiment of overexpressing Pou6f2:The MMQ cells were divided into the control group and experimental group,and the empty plasmid and wild-type Pou6f2 plasmid were transfected into cells respectively according to the instructions of Lipofectamine 3000 transfection kit,and the cells were collected 72 hours later.Western blotting was used to detect the expression of Pou6f2.MTS was used to determine the cell proliferation activity (A490 value),and ELISA was used to determine the level of prolactin (PRL) secretion. Results In the experiment of knocking down Pou6f2,compared with the control group,the expression of Pou6f2 decreased ( P <0.01),the value of A490 increased ( P <0.01),and the level of prolactin secretion increased ( P <0.01) in the experimental group 1 and group 2.There were no significant differences between the experimental group 1 and experimental group 2.In the experiment of overexpressing Pou6f2,compared with the control group,the expression of Pou6f2 increased ( P <0.01),the value of A490 decreased ( P <0.01),and the level of prolactin secretion decreased in the experimental group ( P <0.01). Conclusion Pou6f2 can inhibit the proliferation of MMQ cells and reduce the ability of PRL secretion,which provides a new target for the treatment of prolactinoma.
作者
苗亚洲
谢微嫣
李储忠
张亚卓
MIAO Yazhou;XIE Weiyan;LI Chuzhong;ZHANG Yazhuo(Capital Medical University,Beijing Neurosurgical Institute,Beijing 100070,China)
出处
《山东医药》
CAS
2019年第14期19-22,共4页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81771489)
北京市科学技术委员会资助项目(Z171100000117002)
