摘要
目的:探究慢病毒介导MicroRNA-376b-3p对垂体腺瘤增殖和凋亡的影响及其可能的机制。方法:以体外培养的人垂体腺瘤细胞系为研究对象,分别设立为MicroRNA-376b-3p mimics组(对照组)和慢病毒lenti-sh MicroRNA-376b-3p组(药物组),采用MTT法检测MicroRNA-376b-3p对垂体腺瘤细胞增殖的影响,AnnexinV-FITC/PI双染法检测MicroRNA-376b-3p对垂体腺瘤细胞凋亡率的影响,Western blot法检测MicroRNA-376b-3p对高迁移率蛋白A2(HMGA2)、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白表达的影响。结果:(1)慢病毒介导MicroRNA-376b-3p能够显著抑制垂体腺瘤细胞的增殖(P<0.05);(2)在MicroRNA-376b-3p干预后12 h、24 h以及48 h,垂体腺细胞的凋亡率均明显升高(P<0.05);(3)经MicroRNA-376b-3p转染后,Bax蛋白表达升高,Bcl-2、Caspase-3、Survivin以及HMGA2蛋白表达降低(P<0.05)。结论:慢病毒介导MicroRNA-376b-3p能够明显抑制垂体腺瘤细胞增殖,诱导其凋亡,分析其作用机制可能与下调HMGA2和Survivin表达及上调Bax蛋白表达有关。
Objective:To investigate the effect of lentivirus-mediated microrna-376 b-3 p on the proliferation and apoptosis of pituitary adenomas and its possible mechanism.Methods:Taking human pituitary adenoma cell lines cultured in vitro as the research object,the microRNA-376 b-3 p mimics group(control group)and lenti-sh MicroRNA-376 b-3 p group(drug group)were established respectively,and the MTT method was used to detect MicroRNA-376 b-3 p on the proliferation of pituitary adenoma cells,the AnnexinV-FITC/PI double staining method was used to detect the effect of MicroRNA-376 b-3 p on the apoptosis rate of pituitary adenoma cells,and the Western blot method was used to detect the effect of MicroRNA-376 b-3 p on HMGA2,Bax,Caspase-3 protein expression.Results:(1)Lentivirus-mediated Microrna-376 b-3 p could significantly inhibit the proliferation of pituitary adenoma cells(P<0.05).(2)At 12,24 and48 h after microrna-376 b-3 p intervention,the apoptosis rate of pituitary gland cells in the control group was significantly lower than that in the drug group(P<0.05).(3)After transfection with microrna-376 b-3 p transformation,the expression of Bax protein increased,and the expression of Bcl-2,Caspase-3,Survivin and HMGA2 protein decreased(P<0.05).Conclusion:Lentivirus-mediated Microrna-376 b-3 p could significantly inhibit the proliferation of pituitary adenoma cells and induce apoptosis.The mechanism of microrna-376 b-3 p may be related to its targeted down-regulation of HMGA2 expression,up-regulation of Bax protein expression and regulation of survivin protein expression.
作者
孙凯颉
褚冬
王敏磊
仇诚
李英斌
SUN Kai-jie;CHU Dong;WANG Min-lei;QIU Cheng;LI Ying-bin(Department of Neurosurgery,the Second Affiliated Hospital of Nanjing Medical University,Nanjing,Jiangsu,210000,China)
出处
《现代生物医学进展》
CAS
2020年第22期4225-4228,4217,共5页
Progress in Modern Biomedicine
基金
江苏省中医药科技项目(YB2017093)。
