长链非编码RNA生长停滞特异性转录子5负调控核仁磷酸蛋白1对胃癌细胞顺铂耐药性的影响  

Effect of negative regulation of nucleophosmin 1 by long non-coding RNA growth arrest specific transcript 5 on cisplatin resistance of gastric cancer cells

作　　者：石飞[1] 霍晓雪[1] 徐建国[2] SHI Fei;HUO Xiaoxue;XU Jianguo(Pharmacy Department of Qinghai Provincial People’s Hospital,Xining 810000,Qinghai Province,China;不详)

机构地区：[1]青海省人民医院药学部,青海西宁810000 [2]青海省人民医院肿瘤外科,青海西宁810000

出　　处：《中国生物制品学杂志》2023年第10期1166-1171,1178,共7页Chinese Journal of Biologicals

基　　金：青海省卫生计生指导性课题(2018-wjzdx-12)。

摘　　要：目的 研究长链非编码RNA(LncRNA)生长停滞特异性转录子5(growth arrest specific transcript 5,GAS5)负调控核仁磷酸蛋白1(nucleophosmin 1,NPM1)对胃癌细胞顺铂(cisplatin,DDP)耐药性的影响。方法 选取正常人胃黏膜细胞系GES-1和人胃癌细胞系BG3-823、MGC-803、AGS作为研究对象,qRT-PCR法检测各细胞中LncRNA GAS5水平。诱导AGS细胞DDP耐药(AGS/DDP)后,将实验分为对照组、空质粒组(BC组)、GAS5无义干扰组(pLJM-GAS5NC组)、GAS5过表达组(pLJM-GAS5组),MTT法检测DDP对AGS、AGS/DDP细胞增殖能力的影响;Western blot法检测AGS、AGS/DDP细胞中NPM1、多药耐药基因1 (multidrug resistance 1,MDR1)、耐药基因切除修复交叉互补基因1(excision repair cross complementation group 1,ERCC1)、多药耐药相关蛋白1 (multidrug resistance-associated protein 1,MRP1)、神经性钙黏附素(N-cadherin)水平。结果 与人正常胃黏膜GES-1细胞相比,人胃癌BG3-823、AGS细胞中LncRNA GAS5水平明显降低,其中AGS细胞中LncRNA GAS5水平最低,因此选择AGS细胞进行后续试验。与对照组相比,BC和pLJM-GAS5 NC组AGS细胞中LncRNA GAS5水平明显降低,NPM1、MDR1、ERCC1、MRP1、N-cadherin水平显著升高;与BC组和pLJM-GAS5 NC组相比,pLJM-GAS5组AGS/DDP细胞中LncRNA GAS5水平显著升高,NPM1、MDR1、ERCC1、MRP1、N-cadherin水平显著降低;同浓度DDP(除0μmol/L外)处理后,与对照组相比,BC、pLJM-GAS5 NC组AGS/DDP细胞增殖抑制率明显降低;与BC、pLJM-GAS5 NC组相比,pLJM-GAS5组AGS/DDP细胞增殖抑制率显著升高。DDP对pLJM-GAS5组AGS/DDP细胞48 h的半抑制浓度(IC_(50))为(65.38±5.04)μmol/L,明显低于BC[(120.74±4.17)μmol/L]、pLJM-GAS5 NC组[(120.24±4.29)μmol/L]。结论 上调AGS/DDP细胞中LncRNA GAS5水平,可逆转AGS/DDP细胞耐药性,可能与下调NPM1的表达有关。Objective To investigate the effects of long non-coding RNA(LncRNA) growth arrest specific transcript 5(GAS5) negatively regulating nucleophosmin 1(NPM1) on cisplatin(DDP) resistance of gastric cancer cells.Methods The normal human gastric mucosa cell line GES-1 and human gastric cancer cell lines BG3-823,MGC-803 and AGS were selected as the research objects,of which the level of LncRNA GAS5 in each cell was measured by qRT-PCR.The drug resistance of AGS cells to DDP(AGS/DDP) was induced,and the experiment was divided into control group,empty plasmid group(BC group),GAS5 nonsense interference group(pLJM-GAS5 NC group) and GAS5 overexpression group(pLJM-GAS5 group).MTT method was used to determine the effect of DDP on the proliferation of AGS and AGS/DDP cells;and the levels of NPM1,multidrug resistance 1(MDR1),excision repair cross complementation group 1(ERCC1),multidrug resistance-associated protein 1(MRP1) and N-cadherin in AGS and AGS/DDP cells were measured by Western blot.Results Compared with the normal gastric mucosa GES-1 cells,the level of LncRNA GAS5 in BG3-823 and AGS cells decreased significantly,and among them,the level of LncRNA GAS5 in AGS cells was the lowest,so AGS cells were used for the follow-up experiments.Compared with the control group,the level of LncRNA GAS5 in AGS cells of BC group and pLJM-GAS5 NC group decreased significantly,while the levels of NPM1,MDRl,ERCC1,MRP1 and N-cadherin increased significantly;compared with BC group and pLJM-GAS5 NC group,the level of LncRNA GAS5 in AGS/DDP cells of pLJM-GAS5 group increased significantly,while the levels of NPM1,MDR1,ERCC1,MRP1 and N-cadherin decreased significantly;after treatment with DDP of the same concentration(except 0 μmol/L),compared with the control group,the inhibition rate of AGS/DDP cell proliferation in BC group and pLJM-GAS5 NC group decreased significantly;compared with BC group and pLJM-GAS5 NC group,the inhibition rate of AGS/DDP cell proliferation in pLJM-GAS5group was significantly higher.The semi inhibitory concentrati

关 键 词：长链非编码RNA 生长停滞特异性转录子5 核仁磷酸蛋白1 胃癌 顺铂 

分 类 号：R735.2[医药卫生—肿瘤]