| 贾冬丽,方丽丽,司晓辉,等.小分子干扰RNA抑制黏着斑激酶基因对子宫内膜癌细胞生物学特征的影响[J].安徽医药,2019,23(1):79-83. |
| 小分子干扰RNA抑制黏着斑激酶基因对子宫内膜癌细胞生物学特征的影响 |
| Effect of small interfering RNA silencing of focal adhesion kinase gene on the biological characteristics of endometrial carcinoma cells |
| 投稿时间：2017-04-28 |
| DOI： |
| 中文关键词: 子宫内膜肿瘤 黏着斑激酶1 黏着斑激酶2 RNA,小分子干扰 流式细胞术 印迹法,蛋白质 细胞运动 肿瘤侵润 |
| 英文关键词: Endometrial neoplasms Focal adhesion kinase 1 Focal adhesion kinase 2 RNA,small Interfering Flow cytometry Blotting,western Cell movement Neoplasm invasiveness |
| 基金项目:吴阶平医学基金(320.6750.14109) 小分子干扰RNA抑制黏着斑激酶基因对子宫内膜癌细胞生物学特征的影响贾冬丽1,方丽丽1,司晓辉2,王远菊2 (1.漯河市中心医院妇产科,河南 漯河 462000;2.焦作煤业集团中央医院妇产科,河南 焦作 454150) |
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| 中文摘要: |
| 目的 探讨小分子干扰RNA(small interference RNA,siRNA)抑制黏着斑激酶(focal adhesion kinase,FAK)基因对子宫内膜癌细胞生物学特征的影响。方法 培养人子宫内膜癌HEC-1A细胞,分为siRNA-FAK组、siRNA-阴性对照组和空白对照组,实时荧光定量PCR检测各组细胞中FAK基因表达,细胞增殖能力、迁移和侵袭能力检测分别采用MTT法和Transwell法,蛋白质印迹法(Western blot)检测各组细胞中FAK、磷酸肌醇3激酶(phosphoinositol 3 kinase,PI3K)、磷酸化-Akt(phosphorylate-Akt,p-Akt)、磷酸化-哺乳动物雷帕霉素(phosphorylation-mammalian rapamycin,p-mTOR)蛋白表达。结果 siRNA-FAK组细胞中FAK mRNA和蛋白相对表达量分别为(1.31±0.15)、(0.26±0.08),均低于siRNA-阴性对照组[(2.06±0.18)、(0.62±0.11)]和空白对照组[(2.11±0.20)、(0.64±0.13)],差异有统计学意义(P<0.05)；siRNA-FAK组24 h、48 h、72 h、96 h时细胞吸光度A值均低于siRNA-阴性对照组和空白对照组,均差异有统计学意义(P<0.05)；与siRNA-阴性对照组和空白对照组比较,siRNA-FAK组迁移细胞数和侵袭细胞数均降低,siRNA-FAK组细胞中FAK、PI3K、p-Akt、p-mTOR蛋白表达相对表达量均降低,而Akt和mTOR蛋白相对表达量均升高,均差异有统计学意义(P<0.05)。结论 沉默FAK基因表达可抑制HEC-1A细胞增殖、迁移及侵袭能力,可能与抑制PI3K/Akt/mTOR信号有关。 |
| 英文摘要: |
| Objective To investigate the effect of small interfering RNA (siRNA) silencing of focal adhesion kinase (FAK) gene on the biological characteristics of endometrial carcinoma cells.Methods The human endometrial carcinoma HEC-1A cells were cultured,and all cells were divided into siRNA-FAK group,siRNA-negative control group and blank control group.The expression of FAK gene in each group was detected by using real-time quantitative PCR.The cell proliferation,migration and invasion of cells were detected by using MTT assay and Transwell method.The expressions of FAK,phosphoinositol 3 kinase (PI3K),phosphorylate-Akt (p-Akt) and phosphorylation-mammalian rapamycin (p-mTOR) proteins in each group were detected by Western blot.Results The relative expression levels of FAK mRNA and protein in siRNA-FAK group were (1.31±0.15) and (0.26±0.08),respectively,which were lower than those in the siRNA-negative control group [(2.06±0.18) and (0.62±0.11)] and the blank control group [(2.11±0.20) and (0.64±0.13)],the differences were statistically significant (P<0.05).The absorbance Avalues at 24h,48h,72h and 96h in siRNA-FAK group were lower than those in the siRNA-negative control group and blank control group,the differences were statistically significant (P<0.05).Transwell test results showed that compared with the siRNA-negative control group and blank control group,the number of migrating cells and the number of invasive cells in siRNA-FAK group were decreased,the relative expression levels of FAK,PI3K,p-Akt and p-mTOR proteins in siRNA-FAK group were decreased,while the relative expression levels of Akt and mTOR proteins were increased,the differences were statistically significant (P<0.05).Conclusion Specific silencing of FAK gene could inhibit proliferation,migration and invasion of HEC-1A cells.It might be related to inhibition of PI3K/Akt/mTOR signaling. |
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