Effect of Gecko peptides mixture on proliferation and autophagy in hepatocellular carcinoma SMMC7721 cells
10.13699/j.cnki.1001-6821.2017.09.008
- VernacularTitle:壁虎多肽混合物对肝癌SMMC7721细胞增殖及其自噬的影响
- Author:
Ying JIN
1
;
Ling LIU
;
Leng-Xin DUAN
;
Meng-Li GUO
;
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
- Keywords:
Gecko peptides mixture;
SMMC7721 cell;
autophagy
- From:
The Chinese Journal of Clinical Pharmacology
2017;33(9):798-801
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the possible molecular mechanisms of Gecko peptides mixture (GPM) and research to human hepatocellular carcinoma SMMC7721 cells on autophagy with GPM.Methods SMMC7721 cells were put into plates in its logarithmic phase,and they were treated with different concentration of GPM (0,0.04,0.06,0.09,0.14,0.20,0.30,0.45 mg · mL-1) for 24 h,and then detected corresponding indicators with respective methods.The viability of SMMC7721 cells was detected with 3-(4,5-dimethyl2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT).The concentration of normal group,GPM low-dose,middle-dose and high-dose experimental groups was separately 0,0.1,0.15,0.22 mg · mL-1,according to the results of MTT.Rapamycin was chosen as the positive control drug,which concentration was 40 μg · mL-1.The autophagy of different concentration GPM on SMMC7721 cells was detected by monodansylcadaverine (MDC) staining.Expression levels of Beclin1 and LC3 in SMMC7721 cells were measured by immunohistochemical method and Western blot assay.Results GPM could significantly inhibit the proliferation of SMMC7721 cells in a dose -dependent manner,and the IC50value was 0.16 mg · mL-1 for 24 h.MDC staining showed that there are plenty of autophagosome in cytoplasm,emerging bright green fluorescent in the fluorescence microscope after treatment with GPM for 6 h.The percentage number of positive cells of total cells number in normal group,control group,GPM low,middle and high-dose experimental groups were (15.70 ± 0.26)％,(63.47 ± 0.54)％,(17.09 ± 0.37)％,(70.66 ±0.56) ％,(78.48 ±0.68) ％,respectively.Compared with the normal group,control group,GPM middle and high-dose experimental groups increased,the differences were statistically significant (P ＜ 0.05).The indicators of LC3 in that five groups were (35.84 ± 0.36) ％,(82.41 ± 0.82) ％,(60.83 ± 0.61) ％,(69.66 ± 0.70) ％,(74.61 ± 0.75) ％,respectively.The differences between each group and normal group were statistically significant (P ＜ 0.05).The grayscale average ratio of Beclin1 and β actin in normal group,control group,GPM low,middle and high-dose experimental groups were (6.87 ± 0.68) ％,(11.26 ± 0.87) ％,(6.46 ± 1.34) ％,(11.58 ± 0.95) ％,(15.82 ± 1.58)％,respectively.Compared with the normal group,control group,GPM middle and high-dose experimental groups increased,the differences were statistically significant (P ＜ 0.05).The indicators of LC3 Ⅱ and βactin in that five groups were (26.70 ± 0.41) ％,(103.17 0.88) ％,(30.29 ± 0.52) ％,(36.32 ± 0.52) ％,(64.34 ± 0.48) ％.The differences between each group and normal group were statistically significant (P ＜ 0.05).Conclusion GPM could have an effect on the autophagy in human hepatocellular carcinoma SMMC7721 cells.The possible mechanism may be GPM induced autophagy of SMMC7721 cells,causing cell death ultimately.
