miR21表达与胃癌细胞阿帕替尼敏感性关系的实验研究#br#
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摘要/Abstract

摘要： 目的探讨miR21表达与胃癌细胞对阿帕替尼敏感性的关系以及可能作用机制。
方法体外培养人胃癌AGS细胞和耐药AGSAR细胞，分别转染miR21 inhibitor或miR21 mimics。采用荧光定量PCR（QPCR）检测AGS和AGSAR细胞miR21表达水平；QPCR检测不同浓度（0、2、5、10 μmol／L）阿帕替尼对AGS和AGSAR细胞pVEGFR2和pAkt mRNA表达水平的影响，以及不同miR21表达对AGS和AGSAR细胞pVEGFR2、pAkt、PTEN mRNA表达水平的影响；采用MTT法和划痕实验检测阿帕替尼对AGS、AGSAR细胞增殖和迁移的影响。
结果AGS细胞中miR21、pVEGFR2和pAkt表达量分别为082±009、073±009和068±008，均高于AGSAR的035±010、033±007和025±004，差异有统计学意义（P＜005）；经阿帕替尼处理后，AGSAR细胞的增殖和迁移活性明显高于AGS细胞（P＜005）。10 μmol／L阿帕替尼处理24 h后，阴性对照组AGS细胞的存活率和迁移距离分别为（3948±745）％和（7044±1157）μm，明显低于miR21 inhibitor转染AGS细胞的（8063±827）％和（10846±1039）μm，差异有统计学意义（P＜005）；阴性对照组AGSAR细胞存活率和迁移距离（7882±814）％和（9315±986）μm，明显高于miR21 mimics转染AGSAR细胞的（2995±674）％和（6143±985）μm，差异有统计学意义（P＜005）。转染miR21 inhibitor后，AGS细胞中pVEGFR2和pAkt mRNA相对表达水平分别为112±013和082±010，明显高于阴性对照组的073±011和064±011，而PTEN mRNA表达水平则下调（018±004 vs. 051±008），差异均有统计学意义（P＜005）。转染miR21 mimics后，AGSAR细胞pVEGFR2和pAkt mRNA相对表达水平分别为017±004和012±003，明显低于阴性对照组的030±004和025±002，而PTEN mRNA表达水平则上调（053±006 vs. 044±003），差异均有统计学意义（P＜005）。
结论上调miR21表达可增加VEGFR2和Akt磷酸化水平，从而提高胃癌细胞对阿帕替尼的敏感性。

关键词: 胃癌, microRNA, 阿帕替尼, 敏感性, 血管生成

Abstract: ObjectiveTo investigate the relationship between miR21 expression and sensitivity of gastric cancer cells to apatinib and its possible mechanism.
MethodsAGS cells and drugresistant AGSAR cells were cultured in vitro and transfected with miR21 inhibitor or miR21 mimics respectively. QPCR was used to detect the expression level of miR21 in AGS and AGSAR cells. QPCR was used to detect the effects of apatinib on the expression of pVEGFR2 and pAkt mRNA in AGS and AGSAR cells with different concentrations （0， 2， 5， 10 μmol／L）， as well as the effects of different miR21 expressions on the pVEGFR2， pAkt and pAkt expression levels of AGS and AGSAR cells. The effects of apatinib on proliferation and migration of AGS and AGSAR cells were detected by MTT and scratch test.
ResultsThe expression of miR21， pVEGFR2 and pAkt in AGS cells were 082±009， 073±009 and 068±008 respectively， which were higher than 035±010， 033±007 and 025±004 of AGSAR， and the difference was statistically significant （P＜005）. After the treatment of apatinib， the proliferation and migration activity of AGSAR cells was significantly higher than that of AGS cells （P＜005）. After 10 μmol／L apatinib treatment for 24 h， the survival rate and migration distance of AGS cells in negative control group were （3948±745）％ and （7044±1157） μm respectively， significantly lower than that of miR21 inhibitor transfected AGS cells [（8063±827）％ and （10846±1039） μm]， and the difference was statistically significant （P＜005）. The survival rate and migration distance of AGSAR cells in negative control group （7882±814）％ and （9315±986） μm were significantly higher than （2995±674）％ and （6143±985） μm of AGSAR cells transfected by miR21 mimics， and the difference was statistically significant （P＜005）. After transfection of miR21 inhibitor， the relative expression levels of pVEGFR2 and pAkt mRNA in AGS cells were 112±013 and 082±010 respectively， which were significantly higher than 073±011 and 064±011 in the negative control group， while the expression level of PTEN mRNA was down （018±004 vs. 051±008）， and the differences were statistically significant （P＜005）. After transfection of miR21 mimics， the relative expression levels of pVEGFR2 and pAkt mRNA in AGSAR cells were 017±004 and 012±003 respectively， which were significantly lower than 030±004 and 025±002 in the negative control group， while the expression level of PTEN mRNA was up （053±006 vs. 044±003）， and the difference was statistically significant （P＜005）.
ConclusionUp regulation of miR21 expression can increase the phosphorylation level of VEGFR2 and Akt， thereby enhancing the sensitivity of gastric cancer cells to apatinib.

Key words: Gastric cancer, microRNAs, Apatinib, Sensitivity, Angiogenesis

谭亭昭, 吴超, 王媛, 申振涛. miR21表达与胃癌细胞阿帕替尼敏感性关系的实验研究#br# #br# #br# #br#[J]. 临床肿瘤学杂志, 2018, 23(7): 598-603.

TAN Tingzhao, WU Chao, WANG Yuan, SHEN Zhentao. . Relationship between miR21 expression and sensitivity of gastric cancer cells to apatinib[J]. Chinese Clinical Oncology, 2018, 23(7): 598-603.

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