摘要
目的探讨矿物粉尘诱导基因(mdig)对肺癌细胞顺铂(DDP)敏感性的影响。方法运用实时荧光定量聚合酶链反应和蛋白质印迹法检测A549细胞和DDP耐药的A549细胞(A549/DDP)中mdig mRNA和蛋白的表达水平,进一步分别沉默和过表达A549细胞中的mdig,然后采用CCK-8法检测经不同浓度DDP处理的各组细胞的增殖情况,进而评价其对DDP的药物敏感性变化,并计算DDP的半数抑制浓度(IC50)。结果A549/DDP细胞mdig mRNA和蛋白的表达分别明显高于A549细胞(t=4.8、9.7,P值均<0.01),而且A549/DDP细胞的IC50值明显高于A549细胞[(93.8±8.7)mg/L比(29.9±1.4)mg/L,t=12.6,P<0.01],耐药系数为3.14;A549细胞mdig过表达后,其DDP的IC50值明显增高[LV-mdig组比Vector组为(176.9±13.4)mg/L比(37.6±2.0)mg/L,F=309.0,P<0.01],而mdig沉默后,其IC50值明显降低[LV-mdig RNAi组比LV-con组为(10.8±1.4)mg/L比(26.7±1.1)mg/L,F=146.7,P<0.01]。结论mdig能够降低肺癌细胞对DDP的敏感性,从而导致对DDP的耐药。
Objective To investigate the effect of mineral dust-induced gene (mdig) on the sensitivity of lung cancer cells to cisplatin (DDP). Methods The expression of mdig mRNA and protein in A549 and DDP-resistant A549 cells (A549/DDP) were detected by real-time quantitative PCR and Western blot, then the mdig was further silenced and over-expressed in A549 cells.The CCK-8 assay was used to detect the proliferation of cells treated with different concentrations of DDP, and then to evaluate the sensitivity changes of DDP to calculate the half inhibitory concentration (IC50) of DDP. Results The mRNA and protein expression of mdig in A549/DDP cells were significantly higher than those in A549 cells (t=4.8, 9.7, both P<0.01), and the IC50 value of cisplatin in A549/DDP cells was significantly higher than that in A549 cells [(93.8±8.7) mg/L vs (29.9±1.4) mg/L, t=12.6, P<0.01], drug resistance coefficient was 3.14.Compared with the lentivirus control group, the IC50 value of the mdig-overexpressing A549 cells (LV-mdig) was significantly increased [the LV-mdig group (176.9±13.4) mg/L vs the Vector group (37.5±2.0) mg/L, F=309.0, P<0.01], while the IC50 value of mdig-knockdown A549 cell group (LV-mdig RNAi) was significantly decreased [the LV-mdig RNAi group (10.8±1.4) mg/L vs the LV-con group (26.7±1.1) mg/L, F=146.7, P<0.01]. Conclusions Mdig can reduce the sensitivity of lung cancer cells to cisplatin, resulting in resistance to DDP.
作者
汪庆
耿锋
周昊旻
陈业成
赵洪文
Wang Qing;Geng Feng;Zhou Haomin;Chen Yecheng;Zhao Hongwen(Department of Respiratory and Critical Care Medicine,the First Hospital of China Medical University,Shenyang 110001,China)
出处
《国际呼吸杂志》
2019年第9期649-655,共7页
International Journal of Respiration
基金
国家自然科学资金(81472194)
辽宁特聘教授项目(辽教发[2013]204号).
关键词
肺肿瘤
顺铂
抗药性
肿瘤
矿物粉尘诱导基因
Lung neoplasms
Cisplatin
Drug resistance, neoplas
Mineral dust-induced gene
