| Summary: | 碩士 === 中國醫藥大學 === 中醫學系碩士班 === 99 === Hepatocellular carcinoma (HCC) is ranked fifth in cancer incidence worldwide. In Taiwan, HCC is one of the top two leading causes of cancer death. There is as yet no proven effective medical therapy for treating HCC. Traditional Chinese medicine has been used in cancer treatment or as an adjuvant therapy for cancer treatment for centuries, but there is little scientific evidence on its effectiveness. The aim of this study was to investigate the anticancer effects and molecular mechanisms of a famous formula for patterns of damp heat in liver and gallbladder commonly seen in liver cancer patients - Zuo-Jin-Wan, its components (Coptis chinensis and Evodia rutaecarpa), and its major alkaloidal ingredients (berberine and evodiamine) in human hepatoma HepG2 cells and tumor-bearing mice. In order to use bioluminescence imaging to predict the effect of Zuo-Jin-Wan on hepatic tumor burden in mice. HepG2 cells were stably transfected with a NF-κB luciferase reporter construct to obtain HepG2/NF-κB/luc cells. For the in vitro studies, the treatment of HepG2/NF-κB/luc cells with Zuo-Jin-Wan, Coptis chinensis, Evodia rutaecarpa, berberine, and evodiamine significantly inhibited cell proliferation in a dose- and time-dependent manner by a MTT assay. The values of TC50 after 48 hours treatment were 7.7, 4.3, 320, 5.9, and 0.3 μg/mL, respectively. Then, HepG2/NF-κB/luc cells were treated with Zuo-Jin-Wan, Coptis chinensis, Evodia rutaecarpa, berberine, and evodiamine for 48 hours at TC50 doses and the total RNA was collected for cDNA microarray analysis. Hierarchical cluster analysis revealed that Coptis chinensis shared a similar gene expression profile with Zuo-Jin-Wan, suggesting that Coptis chinensis may be the main component responsible for the anticancer effects of Zuo-Jin-Wan. Network analysis showed that c-myc played a central role in the network topology. For the in vivo studies, the HCC xenograft model in immunocompetent mice was established by direct intrahepatic injection of HepG2/NF-κB/luc cells into ICR mice. Three days after tumor cell implantation, in vivo bioluminescence imaging in mice was performed and mice were assigned into the treatment group (Zuo-Jin-Wan, 200 mg/kg, gavage) and PBS control group. At varying time intervals of 7, 14, or 28 days after daily treatment with Zuo-Jin-Wan or PBS, mice were imaged and immediately sacrificed to examine therapeutic effects of Zuo-Jin-Wan. At 7 days after treatment, statistically significant decreases of tumor ascites fluid and the ratio of tumor weight to liver weight were observed in the Zuo-Jin-Wan group mice compared to control mice. The serum GOT levels in the Zuo-Jin-Wan group were decreased at 7, 14, or 28 days after treatment compared to control mice; however, there was no significant difference. In conclusion, Zuo-Jin-Wan, its components (Coptis chinensis and Evodia rutaecarpa), and its major alkaloidal ingredients (berberine and evodiamine) significantly suppressed tumor growth in HepG2 cells in vitro and in HepG2 tumor-bearing mice. Moreover, c-myc plays a critical role in the anticancer effect of Zuo-Jin-Wan. Our data not only provide scientific evidence for the anticancer effects of Zuo-Jin-Wan on liver cancer but also provide useful guidelines to Traditional Chinese Medicine doctors.
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