Long non-coding RNA CA7-4通过提高细胞自噬水平增强肺癌细胞A594的顺铂耐药

Long non-coding RNA CA7-4 in enhancing cisplatin resistance of lung cancer cell A594 by increasing autophagy levels

目的分析长链非编码RNA(Long non-coding RNA,LncRNA)CA7-4影响非小细胞肺癌(Non-small cell lung cancer,NSCLC)细胞系A549的顺铂(Cisplatin,DDP)耐药的机制。方法将人NSCLC细胞系A549在不同浓度的DDP中培养,检测细胞活力计算半数抑制浓度(Half maximal inhibitory concentration,IC50)。将A549细胞分为对照组、DDP组、CA7-4组及CA7-4+DDP组,其中CA7-4组及CA7-4+DDP组通过转染CA7-4 pcDNA上调LncRNA CA7-4水平;DDP组和CA7-4+DDP组培养基中加入终浓度为12μM的DDP。检测和比较各组细胞的细胞活力、细胞凋亡率和自噬蛋白LC3Ⅰ、LC3Ⅱ水平。结果DDP可剂量依赖性的抑制A549细胞生长,本研究中IC50值为12μM。DDP组的LncRNA CA7-4水平显著低于对照组,CA7-4组的LncRNA CA7-4水平显著高于对照组,并且CA7-4+DDP组的LncRNA CA7-4显著高于DDP组。DDP组细胞活力明显降低而细胞凋亡率显著升高,CA7-4组的胞活力升高、细胞凋亡率降低,并且CA7-4+DDP组的胞活力显著高于DDP组而细胞凋亡率显著低于DDP组。DDP组LC3Ⅱ/LC3Ⅰ水平低于对照组,CA7-4组的LC3Ⅱ/LC3Ⅰ水平高于对照组,且CA7-4+DDP组的LC3Ⅱ/LC3Ⅰ水平显著高于DDP组。结论LncRNA CA7-4可以通过促进NSCLC细胞系A549的自噬促进细胞活力并抑制细胞凋亡,促进细胞对DDP的耐药性。

Objective To analyze the mechanism of long non-coding RNA(LncRNA)CA7-4 in affecting cisplatin(DDP)resistance in non-small cell lung cancer(NSCLC)cell line A549.Methods The human NSCLC cell line A549 was cultured in different concentrations of DDP,and the cell viability was measured to calculate the half maximal inhibitory concentration(IC50).The A549 cells were divided into control group,DDP group,CA7-4 group and CA7-4+DDP group.The CA7-4 group and CA7-4+DDP group were transfected with CA7-4 pcDNA to up-regulate the level of LncRNA CA7-4.DDP group and CA7-4+DDP group were added with DDP at a final concentration of 12μM.The cell viability,apoptosis rate and autophagy protein LC3Ⅰand LC3Ⅱlevels of each group of cells were detected and compared.Results DDP can inhibit the growth of A549 cells in a dose-dependent manner.The IC50 value in this study was 12μM.The level of LncRNA CA7-4 in the DDP group was significantly lower than that in the control group.The level of LncRNA CA7-4 in the CA7-4 group was significantly higher than that in the control group,and the LncRNA CA7-4 in the CA7-4+DDP group was significantly higher than that in the DDP group.The cell viability of the DDP group was significantly decreased and the apoptosis rate was significantly increased.The cell viability of the CA7-4 group was increased,and the apoptosis rate was decreased.The cell viability of the CA7-4+DDP group was significantly higher than that of the DDP group,while the apoptosis rate was significantly lower than that of the DDP group.The level of LC3Ⅱ/LC3Ⅰin the DDP group was lower than that in the control group.The level of LC3Ⅱ/LC3Ⅰin the CA7-4 group was higher than that in the control group.The level of LC3Ⅱ/LC3Ⅰin the CA7-4+DDP group was significantly higher than that in the DDP group.Conclusion LncRNA CA7-4 can promote cell viability and inhibit cell apoptosis by promoting autophagy of NSCLC cell line A549,and promote cell resistance to DDP.