沉默MACC1基因表达对耐药性非小细胞肺癌A549细胞增殖及侵袭的影响

Effect of silencing MACC1 gene on proliferation and invasion of cisplatin-resistant non-small cell lung cancer cells

目的探讨siRNA沉默结肠癌转移相关基因1(MACC1)表达对耐药性非小细胞肺癌细胞株A549增殖及侵袭的影响。方法培养非耐药性和耐药性非小细胞肺癌A549细胞,应用RT-PCR技术检测MACC1 mRNA的表达水平;通过化学合成特异性siRNA,与阳离子脂质体Lipofectamine 2000形成复合体,转染耐药性A549细胞,RT-PCR检测转染效果;继续顺铂药物处理耐药性A549细胞,MTT法检测细胞增殖抑制率;Hoechst33258染色法观察siMACC1后耐药性A549细胞的凋亡形态;Transwell法检测siMACC1后耐药性A549细胞迁移能力;Western blot法检测siMACC1后耐药性A549细胞周期蛋白E-cadherin和N-cadherin的表达水平。结果与耐药性A549细胞组比较,非耐药性A549细胞组中MACC1 mRNA表达量显著下降(P<0.05)。与siRNA NC耐药性A549细胞组比较,siRNA MACC1耐药性A549细胞组中MACC1 mRNA表达量显著降低(P<0.05),细胞增殖抑制率上升(P<0.05);Hoechst 33258细胞染色结果表明siRNA MACC1耐药性A549细胞组发生细胞皱缩,细胞核聚集,细胞形态学呈现细胞凋亡改变;Transwell实验表明MACC1基因沉默后,细胞迁移能力变弱;Western blot结果表明MACC1基因沉默后,与siRNA NC耐药性A549细胞组比较,siRNA MACC1耐药性A549细胞组中E-cadherin蛋白表达显著增加(P<0.05),N-cadherin蛋白表达显著减少(P<0.05),细胞的侵袭转移能力显著减弱。结论 MACC1在耐药性A549细胞中的异常升高,可能是肺癌肿瘤细胞耐药性产生的分子机制之一。siRNA MACCl能够有效转染耐药性A549细胞,能显著地抑制耐药性A549细胞增殖与侵袭。

Objective To observe the effect of down-regulation of MACC1 on cisplatin resistant non-small cell lung cancer(NSCLC) cell line A549 by silencing metastasis associated in colon cancer 1(MACC1) with siRNA interference.Methods Specific siRNA targeting MACC1 was delivered into A549 cells using cationic liposome Lipofectamine 2000.The expression level of MACC1 after transfection was detected by RT-PCR.Hoechst33258 staining was used to investigate the cell growth morphology,and MTT assay was used to detect the changes in the proliferation activity of this cell line after cisplatin treatment.Transwell assay was used to investigate the ability of cells to migrate in vitro.Western blot was used to detect the expression of E-cadherin and N-cadherin protein in cells.Results After the silencing of MACC1 by siRNA,the expression of MACC1 in drug-resistant NSCLC A549 was significantly lower than that in the control group(P<0.05).Compared with the control group,the expression of E-cadherin and N-cadherin protein was significantly lower(P<0.05),and the cell proliferation and migration capacity were significantly lower.Conclusion Silencing MACC1 with siRNA could enhance the efficacy of cisplatin on A549 cells by inhibiting proliferation and promoting apoptosis.