Study on the mechnism of Gecko crude peptides induced apoptosis in KYSE450 cells
10.13699/j.cnki.1001-6821.2017.24.014
- VernacularTitle:壁虎粗多肽诱导食管癌KYSE450细胞凋亡机制研究
- Author:
Meng-Li GUO
1
;
Leng-Xin DUAN
;
Ling LIU
;
Xue-Jiao YANG
;
Ying JIN
;
Yi-Meng DUAN
;
Jian-Gang WANG
Author Information
1. 河南科技大学医学院药理学与分子生物学实验室,河南洛阳471000
- Keywords:
Gecko crude peptide;
KYSE450 cell;
apoptosis;
Caspase-dependent pathway
- From:
The Chinese Journal of Clinical Pharmacology
2017;33(24):2606-2609
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the inhibit proliferation effect of Gecko crude peptide (GCP)on human esophageal carcinoma KYSE450 cells in vitro and to explore its mechanism.Methods KYSE450 cells were put into plates in its logarithmic phase,and they were treated with different concentration (0.10,0.15,0.20,0.25,0.30,0.35,0.4,0.45,0.50,0.55,0.6 mg · mL-1) of GCP for 24,48,72 h,and then detected corresponding indicators with respective methods.The viability of KYSE450 cells was detected with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT').The concentration of blank group(GCP:0),low-dose,middle-dose and high-dose experimental groups(GCP:0.10,0.15,0.22 mg · mL-1)groups were separately,according to the results of MTT.Adriamycin was chosen as the control drug,which concentration was 10 μg · mL-1.After treatment with GCP,the expression of B-cell lymphoma-2(Bcl-2),Bel-2 associated X protein (Bax),cleaved cysteinyl aspartate specific proteinase-3 (Caspase-3),cleaved cysteinyl aspartate specific proteinase-9 (Caspase-9) proteins in KYSE450 cells were observed by Western blot.Results The GCP significantly inhibited proliferation of KYSF450 cells in a dose-and time-dependent manners.After treatment of GCP for 24,48,72 h,the 50％ inhibitory dose (IC50) values were 0.32,0.28,0.24 mg · mL-1,respectively.The Bax and Bcl-2 grayscale average ratio in blank group,low-dose,middle-dose and high-dose experimental groups,control group were (23.83 ± 0.07)％,(51.08 ± 0.78)％,(148.26 ± 6.19) ％,(329.33 ± 4.44) ％,(115.84 ± 0.26) ％.The Caspase-9 and glyceraldehyde phosphate dehydrogenase (GAPDH) grayscale average ratio in above five groups were (14.81 ± 1.05) ％,(20.91 ± 1.24) ％,(39.61 ± 1.89) ％,(43.68 ± 1.13) ％,(39.82 ± 0.69) ％.The Caspase-3 and GAPDH grayscale average ratio in above five groups were (6.37 ±0.51)％,(10.97 ± 0.50)％,(13.44 ± 0.49)％,(17.03 ±0.49)％,(8.63 ± 0.42) ％.Compared with blank group,the differences in drugs groups were significantly (P ＜ 0.05,P ＜ 0.01) Conclusion GCP can inhibit proliferation and induce apoptosis of KYSE450 cells,which may be associated with a Caspase-dependent manner in KYSE450 cells.
