高压氧提高非小细胞肺癌A549细胞对长春瑞滨敏感性的研究

Effect of hyperbaric oxygen on increasing the sensitivity of non-small cell lung cancer A549 cells to vinorelbine

目的探讨高压氧(HBO)联合长春瑞滨(NVB)对非小细胞肺癌A549细胞敏感性的影响及其作用机制。方法人非小细胞肺癌A549细胞培养完成后,按照实验设计分为空白组、HBO组、NVB组及HBO+NVB组,分别不作处理、0.2 MPa HBO处理、IC50浓度NVB处理及0.2 MPa HBO+IC50浓度NVB处理。利用CCK-8实验检测A549细胞增殖能力,流式细胞术检测细胞凋亡情况,Transwell小室实验检测细胞迁移情况,Western blotting实验检测A549细胞相关蛋白的表达水平。结果NVB对A549细胞的IC50值为13.68μmol/L;空白组、HBO组、NVB组及HBO+NVB组的抑制率分别为0、4.10%、46.60%及68.40%,HBO+NVB组抑制率明显高于其他3组(P<0.05或P<0.01);各组凋亡细胞比例分别为3.99%、7.63%、36.92%及56.49%,HBO+NVB组的凋亡率明显高于其他3组(P<0.05或P<0.01)。与其他3组相比,HBO+NVB组A549细胞Bcl-2表达水平明显降低,Bad及Caspase-3的表达水平明显升高(P<0.05或P<0.01);各组抑制迁移细胞比例分别为0、8.90%、42.20%及59.40%,与其他3组比较,HBO+NVB组细胞迁移率最低(P<0.05或P<0.01);与其他3组相比,HBO+NVB组E-Cadherin表达水平明显升高,N-Cadherin表达水平明显降低,P-gp、MRP蛋白的表达水平明显下降(P<0.01)。结论HBO能够提高A549细胞对NVB的敏感性,其作用机制是通过调节耐药、迁移及凋亡相关蛋白来实现的。

Objective To study the effect of hyperbaric oxygen(HBO)combined with vinorelbine(NVB)on the sensitivity of non-small cell lung cancer A549 cells,and to explore its mechanism.Methods After the completion of culture,A549 cells were divided into four groups:blank group,HBO group,NVB group,and HBO+NVB group,with no treatment,0.2 MPa HBO treatment,IC50 NVB treatment,and 0.2 MPa HBO+IC50 NVB treatment,respectively.Cell Counting Kit-8(CCK-8)assay was used to detect the cell prolife-ration.Flow cytometry was adopted to detect cell apoptosis.Transwell experiment was used to detect cell migration.And Western blotting was used to detect the expression changes of A549 cell-related proteins.Results The IC50 value of NVB on A549 cells was 13.68μmol/L.The inhibitory rates were 0,4.1%,46.60%,and 68.40%in the blank,HBO,NVB,and HBO+NVB groups,respectively,and the rate in the HBO+NVB group was significantly higher than those in the other three groups(P<0.05 or P<0.01).The apoptotic rates were 3.99%,7.63%,36.92%,and 56.49%in the blank,HBO,NVB,and HBO+NVB groups,respectively,and the rate in the HBO+NVB group was significantly higher than those in the other three groups(P<0.05 or P<0.01).Compared with the other three groups,the expression level of Bcl-2 in A549 cells in the HBO+NVB group was decreased,while the expression levels of Bad and Caspase-3 were increased(P<0.01).The percentages of cells inhibited from migrating in each group were 0,8.90%,42.20%,and 59.40%,respectively,and the HBO+NVB group had the lowest cell migration rate compared with the other three groups(P<0.05 or P<0.01).Compared with the other three groups,the expression level of E-Cadherin in the HBO+NVB group was increased,while the expression levels of N-Cadherin,P-glycoprotein(P-gp),and Multidrug resistance protein(MRP)were significantly decreased(P<0.01).Conclusion HBO can increase the sensitivity of A549 cells to NVB by regulating the proteins related to drug resistance,cell migration,and cell apoptosis.