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目的:探讨DNA甲基化抑制剂5-氮杂胞苷(5-Aza-2’-deoxycytidine,5-Aza-CdR)在增强人非小细胞肺癌(non-smallcell lung cancer,NSCLC)细胞对吉非替尼敏感性中的作用及其机制。方法:选取EGFR突变型NSCLC细胞株H1650及EGFR野生型NSCLC细胞株H1299,5-Aza-CdR处理后,CCK-8法检测H1650及H1299细胞对吉非替尼敏感性的变化,real-time PCR检测细胞中miR-200c及表皮生长因子受体(epidermal growth factor receptor,EGFR)mRNA的表达水平。结果:EGFR突变型及野生型NSCLC细胞株H1650、H1299对吉非替尼有一定程度的耐药性,5-Aza-CdR处理后H1650及H1299细胞对吉非替尼的敏感性显著增强,表现为吉非替尼对细胞的IC50明显降低[(1.04±0.35)vs(159.37±17.48)μmol/L,(6.28±1.02)vs(223.76±23.63)μmol/L;均P<0.01]。Real-time PCR检测结果显示,5-Aza-CdR处理后EGFR突变型或野生型H1650、H1299细胞中miR-200c[(0.009±0.003)vs(0.002±0.001),(0.004±0.001)vs 0;均P<0.01]和EGFR mRNA[(0.286±0.037)vs(0.015±0.012),(0.057±0.014)vs(0.01±0.01);均P<0.01]的表达水平均显著增高。结论:DNA甲基化抑制剂5-Aza-CdR可上调NSCLC细胞中miR-200c及EGFR mRNA的表达,增强其对吉非替尼的敏感性。
Objective: To investigate the effect of 5-Aza-2-deoxycytidine (5-Aza-CdR), a DNA methylation inhibitor, on the growth of human non-small cell lung cancer (NSCLC) The Role and Mechanism of Fibrinolysin Sensitivity. Methods: The sensitivity of gemcitabine to H1650 and H1299 cells was determined by CCK-8 assay after treatment with EGFR mutant NSCLC cell line H1650 and EGFR wild type NSCLC cell line H1299,5-Aza-CdR. Real-time PCR The expression of miR-200c and epidermal growth factor receptor (EGFR) mRNA in the cells was detected. Results: The EGFR mutant and wild-type NSCLC cell lines H1650 and H1299 showed a certain degree of resistance to gefitinib. The sensitivity of gefitinib to H1650 and H1299 cells after 5-Aza-CdR treatment was significantly enhanced The IC50 of gefitinib was significantly lower than that of the control group [(1.04 ± 0.35) vs (159.37 ± 17.48) μmol / L, (6.28 ± 1.02) vs (223.76 ± 23.63) μmol / L, all P <0.01]. Real-time PCR results showed that miR-200c (0.009 ± 0.003) vs (0.002 ± 0.001), (0.004 ± 0.001) vs 0 in EGFR mutant or wild type H1650 and H1299 cells treated with 5-Aza- (All P <0.01] and EGFR mRNA [(0.286 ± 0.037) vs (0.015 ± 0.012) and (0.057 ± 0.014) vs (0.01 ± 0.01), all P <0.01]. Conclusion: DNA methylation inhibitor 5-Aza-CdR can up-regulate the expression of miR-200c and EGFR mRNA in NSCLC cells and enhance their sensitivity to gefitinib.