We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Regulation of demethylation and re-expression of RASSF1A gene in hepatocellular carcinoma cell lines treated with NCTD in vitro.
- Authors
Yong Wang; Min Xu; Zhen-Hai Di; Jian Zhang; Xue-Qun Mao; Hai-Bo Sun; Wang, Yong; Xu, Min; Di, Zhen-Hai; Zhang, Jian; Mao, Xue-Qun; Sun, Hai-Bo
- Abstract
<bold>Background: </bold>Hepatocellular carcinoma, a lethal malignant neoplasm with poor prognosis, has dismal results of surgical resection and chemoradiotherapy. Norcantharidin (NCTD), the demethylated analog of cantharidin derived from a traditional Chinese medicine, Mylabris, has been used in the treatment of cancer. However, the detailed mechanisms underlying this process are generally unclear. <bold>Purpose: </bold>The aim of this study was to investigate the mechanism of NCTD-induced apoptosis in HepG2 cells. <bold>Materials and Methods: </bold>Human HepG2 cell lines were treated with NCTD at different concentrations (2.50, 5.00, 10.00, 20.00, 40.00 μg/mL) for 24 hours. Cell proliferation was evaluated by measurement of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The methylation levels of RASSF1A (Ras-association domain family 1 A) in HepG2 cells were detected by methylation-specific PCR (MSP). The mRNA levels of RASSF1A in HepG2 cells were detected by real-time fluorescent quantitative PCR (RT-PCR). The levels of RASSF1A protein expression of HepG2 cells were detected by Western blotting assay. <bold>Results: </bold>The inhibition of cell proliferation was observed when treated with NCTD at concentrations (2.5 μg/mL), and as concentration increased, the proliferation of HepG2 cells was markedly inhibited by NCTD in dose-dependent manners. The levels of methylation of RASSF1A decreased at the increasing concentration of 10, 20 and 40 μg/mL. The levels of RASSF1A mRNA and protein were decreased when treated with NCTD at the concentrations of 10, 20 and 40 μg/mL, which were also in a dose-dependent manner. <bold>Conclusion: </bold>NCTD can reverse the methylation state of RASSF1A gene and induce its re-expression, which will provide the theoretical basis for the clinical practice.
- Subjects
LIVER cancer; SURGICAL excision; CHEMORADIOTHERAPY; DNA demethylation; BRUCHUS; CHINESE medicine; CANCER cell proliferation; BROMIDES; PROTEIN metabolism; GENES; HEPATOCELLULAR carcinoma; LIVER tumors; PROTEINS; WESTERN immunoblotting; DNA methylation; CANCER cell culture; IN vitro studies
- Publication
Journal of Cancer Research & Therapeutics, 2015, Vol 11, Issue 4, p818
- ISSN
0973-1482
- Publication type
journal article
- DOI
10.4103/0973-1482.146126