TERT过表达及RNA干扰慢病毒载体对缺氧肺动脉平滑肌细胞增殖影响  

Effects of TERT over-expression and RNA interference lentiviral vector on proliferation of pulmonary artery smooth muscle cells under hypoxia

作　　者：宋景春[1] 徐瑾 刘慧强 曾庆波 钟林翠 胡艳晶 林青伟 余甜 Jing-chun SONG;Jin XU;Hui-qiang LIU;Qing-bo ZENG;Lin-cui ZHONG;Yan-jing HU;Qing-wei LIN;Tian YU(Department of Intensive Care Unit, PLA 94th Hospital, Nanchang 330002, Jiangxi, China)

机构地区：[1]中国人民解放军第九四医院重症医学科

出　　处：《心脏杂志》2019年第4期397-402,共6页Chinese Heart Journal

基　　金：江西省自然科学基金项目资助(20142BAB205009)

摘　　要：目的探讨端粒酶反转录酶(telomerase reverse transcriptase,TERT)过表达及RNA干扰慢病毒载体对缺氧肺动脉平滑肌细胞(pulmonary artery smooth muscle cells,PASMCs)增殖的影响。方法分别构建Plvx-IRES-ZsGreen1-TERT过表达载体和Plvx-shRNA2-TERT干扰载体,培养PASMCs后分为正常对照组、缺氧对照组、空白载体组、TERT过表达组和TERT干扰组,采用CCK-8法进行细胞增殖能力检测,采用流式细胞仪法进行细胞凋亡和细胞周期检测。结果 Plvx-IRES-ZsGreen1-TERT过表达载体和Plvx-shRNA2-TERT干扰载体构建成功后均经测序验证序列正确。缺氧对照组在24 h和48 h时吸光度值较正常对照组明显增加,低氧48 h时G1期细胞比例明显下降,S期细胞比例明显升高,细胞凋亡比例减少(P<0.05);TERT过表达组在低氧24 h和48 h时的吸光度值较缺氧对照组明显升高,低氧48 h时G1期细胞比例较缺氧对照组明显减少,S期细胞比例明显升高(P<0.05);TERT干扰组在低氧24 h和48 h时的吸光度值较缺氧对照组明显减少,低氧48 h时G1期细胞比例较缺氧对照组明显升高,S期细胞比例明显下降,细胞凋亡比率明显升高(P<0.05)。结论低氧时TERT RNA干扰能促进细胞凋亡,减少PASMCs核酸合成,从而抑制PASMCs增殖。AIM To investigate the effects of telomerase reverse transcriptase (TERT) over-expression and RNA interference lentiviral vector on the proliferation of hypoxic pulmonary artery smooth muscle cells (PASMCs). METHODS Plvx-IRES-ZsGreen 1-TERT over-expression vector and Plvx-shRNA 2-TERT RNA interference vector were constructed, respectively. PASMCs were cultured and divided into a normal control group, a hypoxia control group, a blank vector group, a TERT over-expression group, and a TERT RNA interference group. Cell proliferation was detected by CCK-8 method, and apoptosis and cell proliferation were detected by flow cytometry. RESULTS Plvx-IRES-ZsGreen1-TERT over-expression vector and Plvx-shRNA2-TERT RNA interference vector were successfully constructed and sequenced. Compared with those in the normal control group, the absorbance of the hypoxic control group increased significantly, and the proportion of S phase cells increased (P < 0.05). The absorbance of the TERT over-expression group at 24 h and 48 h after hypoxia was significantly higher than that in the hypoxic control group, and G1 phase cells numbers at 48 h after hypoxia were much more than those in the hypoxic control group (P < 0.05). The absorbance value of the TERT interference group significantly decreased at 24 h and 48 h after hypoxia, the proportion of G1-phase cells was significantly increased at 48 h hypoxia, the proportion of S-phase cells significantly decreased, and the apoptosis rate significantly increased (P < 0.05). CONCLUSION TERT RNA interference promotes apoptosis and decreases the synthesis of nucleic acid in PASMCs during hypoxia, thus inhibiting the proliferation of PASMCs.

关 键 词：端粒酶反转录酶 细胞增殖 RNA干扰 肺动脉 平滑肌 

分 类 号：R972.4[医药卫生—药品]