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中国癌症防治杂志

中国癌症防治杂志 ›› 2021, Vol. 13 ›› Issue (3): 242-247.doi: 10.3969/j.issn.1674-5671.2021.03.03

• 基础研究 • 上一篇    下一篇

LncRNA SNHG1靶向miR-101-3p调控胃癌细胞奥沙利铂耐药性

  

  1. 徐州医科大学盐城临床学院，盐城市第一人民医院肿瘤科；上海市公共卫生临床中心病理科 
  • 出版日期:2021-06-25 发布日期:2021-07-07
  • 通讯作者: 宋曙， E-mail：ycss1971@163.com
  • 基金资助:
    2018年盐城市医学科技发展计划项目（YK2018019）

LncRNA SNHG1 targets miR-101-3p to regulate oxaliplatin resistance in gastric cancer cells

  • Online:2021-06-25 Published:2021-07-07

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摘要： 目的 探讨长链非编码RNA(lncRNA)核内小RNA宿主基因1(small nucleolar RNA host gene l，SNHG1)靶向微小RNA-101-3p（miR-101-3p）在胃癌细胞BGC-823中对奥沙利铂(OXA)耐药性的影响及其可能的机制。方法 体外培养BGC-823细胞和耐奥沙利铂细胞BGC-823/OXA，将SNHG1干扰RNA（si-SNHG1组）和阴性对照si-NC（si-NC组）转染耐药细胞BGC-823/OXA，同时设空白对照组（Control组）。转染后再用10 μmol/L奥沙利铂处理，分别记为si-NC+OXA、si-SNHG1+OXA组。采用RT-qPCR检测lncRNA SNHG1、miR-101-3p及MDR1 mRNA的表达；噻唑蓝（MTT）法检测细胞增殖；流式细胞术检测细胞凋亡；Western blot检测P-gp、MRP蛋白表达水平；双荧光素酶报告基因实验验证lncRNA SNHG1与miR-101-3p的靶向关系。结果 与BGC-823细胞相比，BGC-823/OXA细胞中lncRNA SNHG1高表达，miR-101-3p低表达（均P<0.001）。starBase数据库预测显示，lncRNA SNHG1与miR-101-3p 3'UTR区有结合位点。SNHG1-wt+miR-101-3p-mimics组荧光素酶活性低于SNHG1-wt+miR-101-3p-NC组（P<0.001）。与si-NC组相比，si-SNHG1组细胞中lncRNA SNHG1、MDR1及P-gp、MRP蛋白的表达水平及细胞存活率均降低（均P<0.05），细胞凋亡率和miR-101-3p表达水平升高（均P<0.05）；与si-NC+OXA组相比，si-SNHG1+OXA组细胞存活率、P-gp及MRP蛋白表达水平降低，细胞凋亡率升高（均P<0.05）。结论 LncRNA SNHG1通过靶向miR-101-3p抑制胃癌细胞BGC-823增殖和诱导细胞凋亡，增强奥沙利铂耐药性，lncRNA SNHG1可能是胃癌潜在的分子靶点。

关键词: 胃癌, LncRNA SNHG1, miR-101-3p, 奥沙利铂, 耐药

Abstract: Objective To investigate the effect of long non-coding RNA small nucleolar RNA host gene l (SNHG1) targeting microRNA-101-3p (miR-101-3p) on the drug-resistance of oxaliplatin (OXA) in gastric cancer cells (BGC-823) and its possible mechanism. Methods The gastric cancer cells BGC-823 cells and oxaliplatin-resistant BGC-823/OXA cells were cultured in vitro, the SNHG1 interference RNA (si-SNHG1 group) and negative control (si-NC group) were transfected into BGC-823/OXA drug-resistant cells, and the blank control group  was set up at the same time. After transfection, BGC-823/OXA cells were treated with 10 μmol/L oxaliplatin, and classified as the si-NC+OXA group and si-SNHG1+OXA group, respectively. The expressions of lncRNA SNHG1, miR-101-3p and MDR1 mRNA were detected by RT-qPCR; the cell proliferation was detected by MTT assay; the cell apoptosis was detected by flow cytometry; the protein expressions of P-gp and MRP proteins were detected by Western blot; the targeting relationship between lncRNA SNHG1 and miR-101-3p was verified by the double luciferase reporter assay. Results Compared with BGC-823 cells, lncRNA SNHG1 was highly expressed and miR-101-3p was low expressed in BGC-823/OXA cells (both P<0.001). The prediction of starBase database showed that lncRNA SNHG1 had binding sites with miR-101-3p 3'UTR region. The luciferase activity of SNHG1-wt+ miR-101-3p-mimics group was lower than that of SNHG1-wt +miR-101-3p-NC group (P<0.001). Compared with si-NC group, the expression levels of lncRNA SNHG1 mRNA, MDR1 mRNA, P-gp protein, MRP protein and cell survival rate in si-SNHG1 group were decreased ( all P<0.05), and the apoptosis rate and expression of miR-101-3p were increased (both P<0.05). Compared with si-NC+ OXA group, the cell survival rate, protein expression levels of P-gp and MRP in si-SNHG1+OXA group were decreased, and the apoptosis rate was significantly increased (all P<0.05). Conclusions LncRNA SNHG1 inhibits the proliferation of gastric cancer BGC-823 cells and induces apoptosis by targeting miR-101-3p, and enhances oxaliplatin resistance. LncRNA SNHG1 may be a potential molecular target for gastric cancer.

Key words: Gastric cancer, LncRNA SNHG1, miRNA-101-3p, Oxaliplatin, Drug-resistance

中图分类号: 

  • R735.2