上调及下调MicroRNA-195对人脑胶质瘤影响的初步研究  

Effect of microRNA-195 on proliferation of human glioma SHG-44 cells in vivo

作　　者：李晋虎 苗旺[1] 刘晓东[1] 王宏勤[1] 慕伟[1] 王新星[1] 范益民[1] 

机构地区：[1]山西医科大学第一医院神经外科,太原030001

出　　处：《中华临床医师杂志（电子版）》2014年第9期59-62,共4页Chinese Journal of Clinicians(Electronic Edition)

基　　金：山西省基础研究计划(青年科技研究基金)项目(2012021035-4);山西省科技攻关项目(20080311059-1)

摘　　要：目的观察上调及下调microRNA-195(miR-195)对裸鼠皮下荷SHG-44人脑胶质瘤生长的影响并探讨其机制。方法使用脂质体瞬时转染法将miR-195 mimics和inhibitor分别转入人脑胶质瘤细胞系SHG-44,同时设立空白对照组和阴性对照组;实时荧光定量PCR(qRT-PCR)检测转染前后miR-195的含量变化;流式细胞法检测各组细胞周期分布;裸鼠成瘤实验观察miR-195对体内胶质瘤增殖的影响;肿瘤组织HE染色观察病理学改变;Western blot、免疫组织化学染色检测肿瘤组织周期相关蛋白P21、Cyclin D1的表达变化。结果 qRT-PCR测得转染mimics后miR-195的表达水平提高19倍左右,而转染inhibitor后miR-195的表达水平降低为空白对照组的42%;与空白对照和阴性对照组相比,miR-195mimics组细胞阻滞于G0/G1期(P<0.05);而miR-195 inhibitor组则相反(P<0.05);裸鼠成瘤实验显示,miR-195 mimics组与空白对照组和阴性对照组相比,肿瘤生长速度减慢,体积减小(P<0.05),而miR-195 inhibitor组则结果相反(P<0.05);HE染色结果显示miR-195mimics组肿瘤组织异型性下降,新生血管数减少,而miR-195 inhibitor组则相反;Western blot检测示与空白对照组和阴性对照组比较,miR-195 mimics组P21表达上调,而Cyclin D1表达下调,miR-195inhibitor组结果相反(P均<0.05);免疫组化染色检测显示,与空白对照组和阴性对照组比较,miR-195mimics组中P21表达阳性率较高,而Cyclin D1的表达阳性率较低(P均<0.05),miR-195 inhibitor组情况则相反(P均<0.05)。结论 MiR-195可使P21表达升高,下调Cyclin D1的表达,阻滞G0/G1期向S期转换,从而抑制人脑胶质瘤细胞SHG-44的增殖能力。Objective To observe the effects of up-regulated expression and down-regulated expression of miR-195 on the SHG-44 human glioma xenograft growth, and explore the possible mechanism. Methods MiR-195 mimics（Group A） and inhibitor（Group B） were transfected into SHG-44 cell by Lipofectamine RNAiMAX. At the same time, the blank control group（Group C） and negative control group（Group D） were established. MiR-195 expression level was observed by Real-time PCR. Flow cytometry was used to monitor changes in cell cycle. Study the effects of miR-195 on glioma proliferation in vivo by xenograft experiment. Pathological changes of the glioma tissues were observed by HE staining. Western blot and immunohistochemistry were used to detect the expression changes of P21 and Cyclin D1 in removed tumor specimens. Results After transfection of miR-195 mimics, Real-time PCR showed that miR-195 expression level increased about 19 times. However, miR-195 expression level of inhibitor group were decreased to 42.3%of blank control group. Compared with Group C and Group D, cells of Group A were blocked in the G0/G1 phase （P〈0.05）. Group B, however, the result was opposite＆amp;nbsp;（P〈0.05）. Xenograft experiment showed that the volume in Group A was significantly smaller than of those in Group C and Group D （P〈0.05） and the result of Group B was to the contrary （P〈0.05）. HE staining indicated that decreased heteromorphism and reduced number of new vessels in Group A were noted as compared with those in Group C and Group D and the result of Group B was opposite. Western blot showed that the expression levels of P21 in Group A was significantly up-regulated as compared with that in Group C and Group D, while the expression of Cyclin D1 was down-regulated. Group B, however, the situation was opposite. Immunohistochemistry revealed that expression of P21 in Group A was overexpression, while the expression levels of Cyclin D1 was down-regulated （P〈0.05）. However, the result of Group B was opposite

关 键 词：神经胶质瘤 细胞周期 MiR-195 P21 CYCLIN D1 

分 类 号：R739.4[医药卫生—肿瘤]