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Biomed Rep. 2016 Jul;5(1):73-78. Epub 2016 Apr 25.

Mechanism of apoptosis induction in human hepatocellular carcinoma cells following treatment with a gecko peptides mixture.

Author information

1
The Key Laboratory of Pharmacology and Medical Molecular Biology, Medical College, Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China.
2
Department of School Infirmary Pharmacy, South China University of Technology, Guangzhou, Guangdong 510000, P.R. China.
3
Department of Pharmacy, The First Affiliated Hospital of Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China.
4
Department of Pharmacy, The Third Affiliated Hospital of Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China.

Abstract

The aim of the present study was to investigate the apoptotic effect and molecular mechanisms of gecko peptides mixture (GPM) on the human liver carcinoma HepG2 cell line in vitro. The methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay was performed to identify the dose- (0.10, 0.15, 0.20, 0.25 and 0.30 mg/ml) and time-dependent (24, 48 and 72 h) inhibitory effect of GPM on HepG2 cells and their proliferation. Hoechst 33258 staining was carried out to detect the nuclear change coupled with apoptosis induced by GPM. Western blotting was used to evaluate apoptosis-related protein expression changes induced by GPM, including caspase, cytochrome c (Cyt c) and apoptosis-inducing factor (AIF). MTT results showed that GPM significantly inhibited the proliferation of HepG2 cells in a dose- and time-dependent manner. Hoechst 33258 staining demonstrated that GPM induced typical apoptotic morphological changes, while western blotting analysis revealed that GPM increased caspase-3, caspase-9, Cyt c and AIF protein expression levels in HepG2 cells treated with 0.06 or 0.08 mg/ml for 24 h. In conclusion, GPM could induce apoptosis by activating the intrinsic mitochondrial apoptotic pathways.

KEYWORDS:

apoptosis; gecko peptides mixture; human hepatocellular carcinoma; mitochondrial pathway

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