长链非编码RNA NEAT1对胃癌细胞增殖和凋亡的影响及其机制  

The effects and mechanism of long non - coding RNA NEAT1 on cell proliferation and apoptosis in gastric carcinoma cells

作　　者：帅勇锋[1] 王小军[1] 张一中[1] 占大钱[2] Shuai Yongfeng;Wang Xiaojun;Zhang Yizhong;Zhan Daqian(Department of Gastrointestinal Surgery, the Affiliated Hospital of School of Medicine of Ningbo University, Ningbo 315000, China （ Shuai YF, Wang XJ, Zhang YZ;Department of Intensive Care Unit, Tongfi Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China （ Zhan DQ)

机构地区：[1]宁波大学医学院附属医院胃肠外科,315000 [2]华中科技大学同济医学院附属同济医院急危重症科,武汉430030

出　　处：《中华实验外科杂志》2018年第7期1259-1261,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨长链非编码RNAN EAT1（lncRNA NEAT1）对胃癌细胞增殖和凋亡的影响及机制。方法采用实时荧光定量反转录聚合酶链反应（RT-qPCR）测定正常人胃黏膜上皮细胞株GES-1及胃癌细胞株AGS、BSG823、Hs746T和MGC-803中lncRNAN EAT1的相对表达量。将AGS细胞株分为3组，下调表达组（si-NEAT1组）、阴性对照组（si-Ctrl组）及空白对照组（Blank组），细胞毒性活性检测（CCK-8）和克隆形成实验测定细胞增殖，流式细胞术测定细胞凋亡，蛋白印迹测定磷酸化蛋白激酶B（p-Akt）、B细胞淋巴瘤／白血病-2（bcl-2）及bcl-2相关X蛋白（bax）蛋白表达。结果lncRNA NEAT1在胃癌细胞株AGS、BSG823、Hs746T及MGC-803中表达量高于GES-1（P=0．000）。在细胞培养72h及96h后，si-NEAT1组吸光度（A450nm）值低于si-Ctrl组和Blank组（P=0．001）。si-NEAT1组克隆形成率低于si-Ctrl组[（31．43±3．25）％比（67．27±5．81）％，P=0．000]。si-NEAT1组细胞凋亡率高于si-Ctrl组[（19．5±1．2）％比（3．7±0．3）％，P=0．000]。si-NEAT1组p-Akt及bcl-2表达下调，bax表达上调。结论lncRNA NEAT1在胃癌细胞株中高表达，lneRNA NEAT1沉默表达抑制胃癌增殖并诱导凋亡，可能与p-Akt、bel-2下调及bax上调有关。Objective To investigate the effect and mechanism of long non - coding RNA NEATI （IncRNA NEAT1 ） on proliferation and apoptosis of gastric carcinoma cells. Methods The expression level of lneRNA NEAT1 and normal gastric cell line GES - 1 was measured by real - time quantitative reverse transcriptase - polymerase chain reaction （ RT - qPCR）. The AGS cell line was divided into three groups, si - NEATI group, si -Ctrl group and Blank group. The proliferation ability was measured by methyhhiazolyldipheuyl- tetrazolium bromide （MTF） assay and clone formation assay. Apoptosis ability was tested by flow cytometry. The expression level of Phosphor - protein KinaseB （ p - Akt） , B cell lym- phoma/leukemia- 2 （bcl- 2 ） and bcl -2 associated X protein （bax） protein was measured by Western blotting. Results The expression level of IncRNA NEAT1 in AGS, BSG823, Hs746T and MGC - 803 cell line was significantly higher than that in GES- 1 （P = 0. 000）. CCK- 8 demonstrated that the A450 .,, of si - NEAT1 group was significantly lower than that in si -Ctrl and Blank group alter culturing for 72 and 96 h （ P = 0. 001 ）. The number of clone formation cells in si - NEATI group was significantly lower than that in si-Ctrl group, （31.43＋3.25）% vs. （67.27＋5.81）%, P=0.000. The apoptosisrateofsi- NEAT1 group was significantly higher than that in si - Ctrl group, （ 19. 5 ± 1.2） % vs. （ 3.7 ± 0. 3 ） %, P = 0. 000. The expression level of p - Akt and bcl - 2 protein in si - NEAT1 group was significantly up - regulated, while bax down - regulated. Conclusion LncRNA NEAT1 was over - expressed in gastric carcinoma cell lines. Knockdown of lncRNA NEAT1 could inhibit the proliferation and induce apoptosis, possibly through down - regulation of p - Akt and bcl - 2 protien, and up - regulation of bax.

关 键 词：长链非编码RNA NEAT1 胃癌 增殖 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]