| Summary: | 博士 === 國立陽明大學 === 傳統醫藥研究所 === 106 === Background and Purpose: In clinical practice, the esophageal cancer and hepatocellular carcinoma (HCC) are difficult in to be diagnosed in early stage and have poor prognosis in unresectable stages. Resistant to treatment and prone to metastasis are the major causes of dismal survival. Cancer stem cells (CSCs) are regarded as the basis for tumor initiation, development, treatment resistance, recurrence and metastasis. Antrodia cinnamomea, also known as niu-chang-chih, is a fungus indigenous to Taiwan which grows on decayed Cinnamomum kanehirae. In folk medicine, it is well known and widely used for antidotal and anti-tumor therapy. To avoid contamination in the wild source and to expand the production of this rare fungus for medical use, A. cinnamomea mycelial fermentation broth (AC-MFB) produced by a novel biotechnology has been developed by our group. The purpose of this study was to investigate the effects of AC-MFB on cancer and clinical characteristics of CSCs.
Materials and Methods: HA22T/VGH human hepatic cancer cells (HCCs), 1MEA.7R.1 mice HCCs, CE81T/VGH human esophageal cancer cells (ESCs) cells, BE3 ESCs, EA. hy926 and SVEC4-10 endothelial cells (ECs) were used in this study. The CSCs prosperity of the HCCs and ESCs cells and the effects of AC-MFB on CSCs characters including sphere formation, angiogenesis, DNA repair, and epithelial mesenchymal transition (EMT) were evaluated.
Results: The HA22T HCCs, CE81T/VGH and BE3 ESCs cells possessed spheres-forming activity, indicative of properties of CSCs. The in vitro study showed AC-MFB could inhibit cellular viability of HCCs and ESCs in both dose-dependent and time-dependent manners. The colony formation assay showed that pretreatment with AC-MFB decreased the survival of irradiated esophageal cancer cells, with a maximum sensitizer enhancement ratio of 1.91 at 37% survival. As for morphological alteration, TGF-β triggered BE3 ESCs cells from polygonal to spindle shape which was reversed by AC-MFB. Migration of BE3 ESCs cells, a hallmark of EMT for invasiveness, was augmented by TGF-β, and it could be inhibited by AC-MFB treatment. The in vivo study showed AC-MFB could inhibit the growth of HCC and enhance the radiation effect on esophageal tumor growth delay in mice without liver function abnormality. In anti-angiogenesis assay, AC-MFB could inhibit the cellular viability, migration, and tube formation activity of EA. hy926 and SVEC4-10 ECs. The production of extracellular vascular endothelial growth factor (VEGF) and intracellular hypoxia-inducible factor-1 alpha (HIF-1α) from HCC cells was also suppressed by AC-MFB. The immunofluorescence assay and a Western blotting assay showed that AC-MFB could delay the abrogation of γ-H2AX, an indicator of DNA double-strand breaks. The radiosensitizing effect of AC-MFB was accompanied by the upregulation of p21 expression and downregulation of the radiation-induced phosphorylation of ataxia telangiectasia-mutated kinase and checkpoint kinase 2. The results also showed AC-MFB treatment could inhibit the TGF-β-induced overexpression of Vimentin; reverse the E-cadherin to N-cadherin switch in esophageal cancer BE3 cells. The anti-EMT effect of AC-MFB was accompanied by the downregulation of Twist.
Conclusion: The modern biotechnological product of indigenous medicinal fungus Antrodia cinnamomea possesses bioactivity against CSCs characters of esophageal cancer and HCC by multiple molecular pathway modulation.
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