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

Jin, Ying; Duan, Leng‐Xin; Xu, Xinli; Ge, Wenjing; Li, Ruifang; Qiu, Xiangjun; Song, Ying; Cao, Shanshan; Wang, Jiangang

doi:10.3892/br.2016.664

Cited by 12 publications

(7 citation statements)

References 29 publications

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“…The cells were treated with five different concentrations of each peptide ranging from 0.0015 µM to 15 µM for a total of 24 h before viability measurements were performed. The results show that the peptides did not have any major inhibitory effect on the growth or viability of HEK-293 cells over the majority of concentrations tested, however the full length peptides induced 20–40% loss in viability at the highest concentration tested (Figure 6) [42,43,44], with AP3K showing the highest extent of cytotoxicity (~40% dead cells) at 15 µM peptide, while both AP3 and AP3K exhibited some cytotoxic activity (~18% dead cells) at 1.5 µM peptide.…”

Section: Resultsmentioning

confidence: 99%

Characterization and Antimicrobial Activity of Amphiphilic Peptide AP3 and Derivative Sequences

2019

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The continued emergence of new antibiotic resistant bacterial strains has resulted in great interest in the development of new antimicrobial treatments. Antimicrobial peptides (AMPs) are one of many potential classes of molecules to help meet this emerging need. AMPs are naturally derived sequences, which act as part of the innate immune system of organisms ranging from insects through humans. We investigated the antimicrobial peptide AP3, which is originally isolated from the winter flounder Pleuronectes americanus. This peptide is of specific interest because it does not exhibit the canonical facially amphiphilic orientation of side chains when in a helical orientation. Different analogs of AP3 were synthesized in which length, charge identity, and Trp position were varied to investigate the sequence-structure and activity relationship. We performed biophysical and microbiological characterization using fluorescence spectroscopy, CD spectroscopy, vesicle leakage assays, bacterial membrane permeabilization assays, and minimal inhibitory concentration (MIC) assays. Fluorescence spectroscopy showed that the peptides bind to lipid bilayers to similar extents, while CD spectra show the peptides adopt helical conformations. All five peptides tested in this study exhibited binding to model lipid membranes, while the truncated peptides showed no measurable antimicrobial activity. The most active peptide proved to be the parent peptide AP3 with the highest degree of leakage and bacterial membrane permeabilization. Moreover, it was found that the ability to permeabilize model and bacterial membranes correlated most closely with the ability to predict antimicrobial activity.

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Section: Resultsmentioning

confidence: 99%

Characterization and Antimicrobial Activity of Amphiphilic Peptide AP3 and Derivative Sequences

2019

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“…In the past decade, many traditional Chinese medicines have been applied as anti-tumor agents, and have been characterized by low toxicity and promising therapeutic effects 7 . For instance, the gecko ethanol extract (GEE) has been proven to inhibit the growth of H22 xenograft tumors in vivo and induce apoptosis in vitro 8 . Other studies have demonstrated that GEE can inhibit SiHa cell growth, which may be through the inhibition of cell proliferation, angiogenesis and survival.…”

Section: Discussionmentioning

confidence: 99%

“…Consequently, the high efficiency and low toxicity of natural anti-tumor medicines extracted from plants and animals may be a new strategy to treat cancer 7 . Gecko extracts, has been shown to inhibit the growth of various tumor cells, such as SiHa, EC109, SMMC7721 cells 8 , 9 . With the development of genomics, transcriptomics, proteomics, and metabolomics technologies, RNA-seq technology has been widely used in recent years 10 .…”

Section: Introductionmentioning

confidence: 99%

Differentially expressed genes of HepG2 cells treated with gecko polypeptide mixture

et al. 2018

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Gecko (Gekko japonicus) extracts have been used in traditional Chinese medicine for many years. It has been proven that the gecko polypeptide mixture (GPM) extracted from gecko can inhibit the growth of multiple types of tumor cells. In order to investigate the possible anti-tumor molecular mechanisms of GPM, we used RNA-seq technology to identify the differentially expressed genes (DEGs) of human hepatocellular carcinoma (HCC) HepG2 cells treated with or without GPM. MTT assay was used to detect the viability of HepG2 cells. DAPI fluorescence staining was performed to observe morphological changes in the nuclei of HepG2 cells. Western blot analysis was applied to observe the expressions of apoptosis-related and endoplasmic reticulum stress (ERS)-related proteins in HepG2 cells. Flow cytometry assay was performed to detect the apoptosis and reactive oxygen species (ROS) in HepG2 cells. Our results showed that GPM inhibited HepG2 cells proliferation and induced the apoptosis of HepG2 cells. RNA-seq analysis suggested that the ER-nucleus signaling pathway involved in the anti-cancer molecular mechanism of GPM. Therefore, GPM may induce apoptosis in HepG2 cells via the ERs pathway.

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“…The results of both real-time PCR and immunohistochemistry were the same for p53 expression. On the other side, caspase-3 gene expression was not detected by real-time PCR, and this may be explained by the late expression of caspase-3 in tumor cells (Jin et al, 2016) that could by detected immunohistochemically using a speci ic antibody to the expressed protein in the localized tissue.…”

Section: Immunohistochemical Analysismentioning

confidence: 99%

Effect of Interferon-Beta (IFN-β) on tumor suppressor and apoptotic markers in hepatocellular carcinoma cell line

Okasha

Hassan

Aboushousha

et al. 2019

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IFN-β (Rebif) is a drug with valuable cancer therapeutic potential as it mediates anti-proliferation and apoptosis induction. Therefore, this study aimed to evaluate the anti-proliferation of IFN-β through assessing gene expression of p53 and caspase-3 in hepatocellular carcinoma cell (HCC) line (HepG2). The 50% inhibition viability concentration (IC50) of IFN-β was calculated by cytotoxicity assay, and it was tested on normal cell line (Vero). After treating HepG2 cells with IFN-β, p53 and caspase-3 expression was analyzed, at time intervals of 2, 4, 6, and 24 hrs, by real-time PCR, histopathology and immunohistochemistry. IC50 of IFN-β was 420 µg/ml, which wasn't cytotoxic on normal cells. P53 expression was gradually up-regulated by time, and then, it was decreased after 24 hrs incubation. However, no expression of caspase-3 was detected compared to cell control. Histopathologically, cells degeneration was remarkable after 24 hrs while no change was noticed in control. Immunohistochemical analysis revealed a decline of p53 and caspase-3 expression in treated cells with IFN-β, after 24 hrs, to 30% compared to cell control (50% and 60%, respectively). IFN-β has the potential to be utilized as a suppressor of HCC proliferation and inducer of malignant cells apoptosis.

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

Cited by 12 publications

References 29 publications

Characterization and Antimicrobial Activity of Amphiphilic Peptide AP3 and Derivative Sequences

Characterization and Antimicrobial Activity of Amphiphilic Peptide AP3 and Derivative Sequences

Differentially expressed genes of HepG2 cells treated with gecko polypeptide mixture

Effect of Interferon-Beta (IFN-β) on tumor suppressor and apoptotic markers in hepatocellular carcinoma cell line

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