Abstract
Purpose : To evaluate the antiproliferative and apoptotic effects of high-dose vitamin C in cholangiocarcinoma cell line (CCA). Methods : Sulforhodamine B colorimetric assay was used to determine cell proliferation in human K100- CCA cell lines. The expression of anti-apoptotic Bcl-2 protein/ pro-apoptotic Bax protein was evaluated by Western blot analysis. Results : Ascorbic acid inhibited the growth of CCA in a dose-dependent manner. The half-maximal inhibitory concentration (IC 50 ) of ascorbic acid on K100 at 24 h was 55 ± 9.4 mM. It was observed that treatment with ascorbic acid resulted in the reduction of glutathione and increased hydrogen peroxide contents in the cells. Moreover, disturbance of mitochondrial membrane potential occurred in ascorbic acid-treated cells in a dose-dependence manner. The ratio of Bcl2/Bax decreased in CCA treated with ascorbic acid. Conclusion : The results show that the anti-proliferation effect of ascorbic acid in CCA may be attributable to the modulation of expressions of Bcl-2 and Bax proteins and dissipation of the mitochondrial electrochemical potential gradient, which is an early event leading to apoptosis. Keywords : Vitamin C, Cholangiocarcinoma, Apoptosis, Glutathione, Mitochondrial membrane potential
Highlights
Cholangiocarcinoma (CCA) is a rare form of cancerous tumour that originates from the biliary epithelial cells
The effect of ascorbic acid on cell proliferation was evaluated by sulforhodamine B (SRB) assay in K100 cholangiocarcinoma cell line (CCA) cell line (Figure 1)
Treatment with ascorbic acid led to the rapid depolarisation of ∆Ψm, which was marked by green fluorescence of JC-1 monomeric forms presented in the cytosol (Figure 4)
Summary
Cholangiocarcinoma (CCA) is a rare form of cancerous tumour that originates from the biliary epithelial cells It is prevalent in the Southeast Asian region and is associated with symptoms like liver fluke infection (caused due to Opisthorchis viverrini and Chlonorchis sinensis). Several studies have reported using a high dose of ascorbic acid for treating cancer [5,6,7,8,9]. A high dose of pharmacological ascorbic acid concentration was found to induce in vitro cell death and in vivo tumour growth inhibition [12]. The present study aimed at evaluating the antiproliferative and apoptotic effects of highdose vitamin C or ascorbic acid treatment on K100 CCA cell line. The cells were pre-treated with ascorbic acid (0.5 - 100 mM) for 24 h.
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