长链非编码RNA LOC101928316对胃癌细胞BGC-823细胞周期和靶基因的调控  

Role of long non-coding RNA LOC101928316 in cell cycle and target genes regulation in gastric cancer BGC-823 cells

作　　者：张雯雯 杨圣 姚文卓 马海涛 胡逢静 郭惠珍 曾勇 王俊玲[1] 梁戈玉 李成云[1] ZHANG Wen-wen;YANG Sheng;YAO Wen-zhuo;MA Hai-tao;HU Feng-jing;GUO Hui-zhen;ZENG Yong;WANG Jun-ling;LIANG Ge-yu;LI Cheng-yun(Department of Toxicology,School of Public Health,Lanzhou University,Lanzhou Gansu 730000,China;Key Laboratory of Environmental Medicine Engineering,Ministry of Education,School of Public Health,Southeast University,Nanjing Jiangsu 210009,China)

机构地区：[1]兰州大学公共卫生学院卫生毒理学研究所,甘肃兰州730000 [2]东南大学公共卫生学院环境医学工程教育部重点实验室,江苏南京210009

出　　处：《毒理学杂志》2020年第2期103-108,共6页Journal of Toxicology

基　　金：国家自然科学基金(81803188,81673132);兰州大学中央高校基本科研业务费专项资金。

摘　　要：目的探讨长链非编码RNA(long non-coding RNA, lncRNA)LOC101928316对胃癌BGC-823细胞周期及靶基因的调控作用。方法通过慢病毒对BGC-823细胞稳定转染LOC101928316,RT-qPCR法验证转染效果。运用MTT检测LOC101928316转染对BGC-823细胞增殖的影响;流式细胞术检测LOC101928316转染对BGC-823细胞周期和凋亡的影响;使用KEEG数据库对LOC101928316相关基因进行生物信息学分析,探讨其潜在调控的靶基因;应用RT-qPCR和Western blot检测LOC101928316对靶基因和蛋白表达的调控。结果与阴性对照组相比,LOC101928316过表达稳定转染可引起BGC-823细胞增殖能力降低(P<0.05);细胞周期出现S期阻滞(P<0.05),且细胞凋亡率升高,差异有统计学意义(P<0.05)。LOC101928316主要参与PI3K信号通路的调控,AKT3和PTEN基因为其潜在调控的靶基因。与阴性对照组相比,LOC101928316过表达转染可引起BGC-823细胞p-AKT3蛋白表达降低(P<0.05);PTEN基因mRNA表达和蛋白表达均升高(P<0.05)。结论 LOC101928316可能通过抑制PI3K信号通路而引起胃癌细胞的细胞周期阻滞,进而负向调控胃癌细胞的增殖。研究结果为LOC101928316在胃癌中的功能发挥和调控机制研究提供一定的线索。Objective To investigate the regulation of cell cycle and target genes in gastric cancer BGC-823 cells by long non-coding RNA(lncRNA) LOC101928316. Methods Lentiviral stable transfection was used to up-regulate the lncRNA LOC101928316 expression level in gastric cancer cell line BGC-823. Then, MTT assay and flow cytometry were used to detect the effect of LOC101928316 on cell proliferation, cell cycle and apoptosis of BGC-823 cells. Regulation relationships of LOC101928316 was searched by KEEG tool to predict the possible signaling pathways and potential target genes. RT-qPCR and Western blot were used to detect the LOC101928316 regulation on both mRNA and protein expression levels of its predicted target gene. Results Compared with the negative control group, LOC101928316 stably-transfected group caused BGC-823 cells proliferation decreased(P<0.05), caused BGC-823 cell cycle arrest in S phases(P<0.05), and increased BGC-823 cells apoptosis rate(P<0.05). LOC101928316 is mainly involved in the regulation of PI3 K signaling pathway, and AKT3 and PTEN genes are important potential target genes. Compared with the negative control group, up-regulated LOC101928316 expression in the transfected group decreased p-AKT3 protein expression(P<0.05), and PTEN mRNA and protein expression were both increased(P<0.05). Conclusion lncRNA LOC101928316 may induce the cell cycle arrest of BGC-823 and negatively regulates the proliferation of gastric cancer cells by regulating PI3 K signaling pathway. The result help us to further understanding the role and mechanism of LOC101928316 in gastric cancer cells.

关 键 词：LOC101928316 BGC-823细胞 细胞周期 基因调控 

分 类 号：R735.2[医药卫生—肿瘤]