Effect of Withaferin A on epithelial-mesenchymal transition and apoptosis in cells of lung squamous cell carcinoma
10.3760/cma.j.issn.1006-9801.2018.12.002
- VernacularTitle:醉茄素A对肺鳞状细胞癌细胞上皮间质转化及凋亡的影响
- Author:
Pengfei WANG
1
;
Fengjuan LIU
;
Xiao WU
;
Rongli SUN
Author Information
1. 266042,青岛大学第二临床医学院呼吸与危重症医学科
- Keywords:
Withaferin A;
Squamous cell carcinoma,lung;
Apoptosis;
Epithelial-mesenchymal transition
- From:
Cancer Research and Clinic
2018;30(12):825-829
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the regulation of Withaferin A on apoptosis-related proteins in lung squamous cell carcinoma and its effect on cell epithelial-mesenchymal transition.Methods Lung squamous cell carcinoma cell line SK-MES-1 were cultured in vitro.The SK-MES-1 cells were treated with the final mass concentration of 0 (control),5,10,20,and 40 μg/ml for 28 h,and the general morphology and shedding of the cells were observed under phase contrast microscope;MTT assay was used for detection of cell viability;flow cytometry was used for detection of apoptosis;immunofluorescence and real-time fluorescent polymerase chain reaction (RT-PCR) was used for detection of apoptosis-related proteins and genes bcl-2 and bax,and expression of the epithelial marker E-cadherin and the interstitial marker Vimentin.Results Different concentrations (0,5,10,20,40 μg/ml) of Withaferin A inhibited the activity of SK-MES-1 cells with cell viability of 0.62±0.05,0.42±0.04,and 0.33±0.06,0.21±0.03,0.17±0.04,respectively,which suggesting that this inhibition was related to the concentration of Withaferin A (F =386.505,P =0.005).After treatment with SK-MES-1 cells for 24 h at different concentrations (0,5,10,20,40 μg/ml) of Withaferin A,the apoptosis rates of each group were (0.180±0.011) ％,(0.310±0.013) ％,(0.500±0.021) ％,(0.540±0.018) ％,and (0.410± 0.027) ％,which suggesting that Withaferin A rarely caused apoptosis,mostly necrotic cells (F =1 065.78,P =0.124).In SK-MES-1 cells treated with different concentrations of Withaferin A for 24 h,the results of immunofluorescence showed that the expression of Vimentin was decreased in the experimental group at a concentration of 20 μg/ml compared with the control group,and the fluorescence intensity was lower than that of the control group,but the fluorescence intensity of E-cadherin was higher than that of the control group;the intensity did not change significantly in the experimental group with the other concentrations.While the expression levels of bax and bcl-2 proteins in the control group and the experimental group did not change significantly.RT-PCR results showed that the mRNA expression of E-cadherin (6.7±0.6 and 6.4±0.9) in the experimental group at a concentration of 20 and 40 μg/ml was significandy higher than that in the control group (4.2±1.0),and the difference was statistically significant (both P ＜ 0.05),and the mRNA expression of Vimentin (4.7±0.5 and 4.7±0.5) was significantly lower than that in the control group (7.2±0.7),and the difference was statistically significant (both P ＜ 0.05).Conclusion Withaferin A can inhibit the growth of lung squamous cell carcinoma cells and inhibit the process of epithelial-mesenchymal transition,but it has no obvious relationship with apoptosis and apoptosis-related proteins.
