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Vol.9 No.3-2:Biomolecular evaluation of apoptosis, cell cycle, oxidative stress, and limiting enzymes of the glycolytic pathway in hepatocellular carcinoma cell line HepG2 treated with crude snake venom with or without sorafenib

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By: Maha Y. Abdel Al Shakour1, Emad M. Elzayat2*, Khalid M. Mahmoud3,

Mamdouh I. Nassar4, Abdel Hamid Z. Abdel-Hamid5

1PhD student at Zoology Department, Faculty of Science, Cairo University, Giza, Egypt

2* Biotechnology Department, Faculty of Science, Cairo University, Giza, Egypt

3Pharmacognosy Department, NRC, Giza, Egypt

4Entomology Department, Faculty of Science, Cairo University, Giza, Egypt

5Therapeutic Chemistry Department, NRC, Giza, Egypt

Abstract

Background: Natural venoms have biological activities including anti-inflammatory, antimicrobial, and anticancer effects.  Hepatocellular carcinoma (HCC) is still a worldwide problem and difficult to treat by chemotherapeutic agents especially sorafenib (SOR), as it evokes many harsh side effects and is disable to differentiate between normal and cancer cells. Objective: The present study aimed to test the hypothesis that combining crude venoms of the snake or the bee or the scorpion could synergistically enhance the antiproliferative effects of SOR in hepatocellular carcinoma cell line (HepG2). Experimental design: Separate crude venoms have been applied to HepG2 cells and normal human retinal cells(RBE1) for estimation of IC50. The most effective venom has been combined with sorafenib in five nonconstant ratios and the combination index (CI) was estimated to expose their synergistic or antagonistic action. The best combination was used for downstream analysis. Results: The crude snake venom exhibited the most cytotoxic effect and the least IC50. It has been combined with sorafenib, and the combination index (CI) was calculated. IC25 SV + IC10 SOR was the best combination with CI=0.209 indicating high synergistic cytotoxic activity against HepG2. The underlining molecular mechanisms of action, in terms of the expression level of apoptotic genes (p53, Bax, Caspase 3, and Bcl2), flow cytometric analysis of cell cycle, oxidative stress markers as well as the activity of some limiting enzymes in the glycolytic pathway (ALDOB, PK and LDH) have been investigated.

Conclusion: Our results suggest a novel synergistic, and anti-proliferative effect of snake venom with sorafenib on HepG2 cells.

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Vol.5 No.2 – 9 : Berberine attenuates cancer cell growth via modulating the cell cycle dynamics but not apoptosis in human colorectal HCT-116 3D spheroid model

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By: Ahmed A. Soffar

Division of Molecular Biology, Department of Zoology, Faculty of Science,

Alexandria University, Alexandria, Egypt

Abstract

Colorectal carcinoma is a cosmopolitan type of cancer with poor prognosis, motivating seeking novel strategies to prevent disease development and progression. The poor prognosis is attributed to the severe toxic side effects of the current therapeutic regimes. Hence, novel less toxic treatment strategies are urgently warranted. Berberine is a natural compound with several biological and pharmacological properties, including anti-fungal, anti-diabetic, cardioprotective effects. Some reports showed that berberine inhibits cell growth by inducing cell cycle arrest and promotion of apoptosis in cancer cells. Importantly, the anticancer potential of berberine in colorectal cancer has not been previously investigated. Hence, this work aims to investigate whether berberine possess anticancer properties against colorectal HCT116 cancer cells. The potential effect of berberine on cell cycle regulation and apoptosis will also be deeply investigated. This work was conducted using the more physiological 3D spheroid culture model that mimics better the impact of the tumor microenvironment as well as the cell-cell interaction in the cellular response to therapy. When compared to the previous studies, this work will explain the mode of action of berberine in more physiological conditions that better mimics the in vivo situation. In order to achieve the goal of this work, spheroid growth assay as well as proliferation assay were performed. Spheroid cell suspensions were further investigated using flow cytometry to assess the cell cycle distribution of cells upon berberine application. BrdU immunostaining was performed to elucidate the S-phase fraction of cells. The proliferation potential and the level of apoptosis were also investigated by Ki67 and Annexin V labelling, respectively. The results showed that berberine attenuated tumor spheroid growth and limits the proliferative capacity of HCT116 cells. This could be attributed to the berberine-mediated G1-phase cell cycle delay. The S-phase fraction of cells was significantly decreased upon berberine application. Unexpectedly, berberine did not induce a significant difference in the % of apoptotic cell fraction of cells as compared to the controls. Collectively, these results suggest that berberine possesses an anti-tumor efficacy in 3D culture preparations via modulating the cell cycle progression. Specifically, berberine induces G1-phase cell cycle delay and decreases the S-phase fraction of cells. Thus, it limits the proliferative capacity of cells. Also, berberine did not induce programmed cell death in the HCT116 spheroids.


Berberine attenuates cancer cell growth via modulating the cell cycle dynamics but not apoptosis in human colorectal HCT-116 3D spheroid model-converted

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