Cholangiocarcinoma (CCA) is a cancer that arises in the epithelium of biliary tree, both intra-hepatic and extra-hepatic bile ductsdence of this cancer is relatively rare than are other gastrointestinal cancers (Shaib and El-Serag, 2004). However, high prevalence has been recorded in the Southeast Asian region, in particular the north-eastern part of Thailand (Sripa and Pairojkul, 2008). The mortality rate of CCA patients is high due to late clinical presentation. The primary treatment strategy for CCA is surgical resection of the tumor. However, only some cases are applicable for surgery, posing a limitation of the therapeutic options. The 5-year survival rate after resection is very low (20–30%) (Rosen et al., 2010). Most conventional chemotherapeutic agents such as gemcitabine, cisplatin, paclitaxel, and oxaliplatin have very limited efficiency in CCA patients and also could induce chemo-resistance (Lee et al., 2008 Valle et al., 2010 Weigt and... See full list on Chemicals and Antibodies Andrographolide (98% purity), ribonuclease A, and phenylmethylsulfonyl fluoride (PMSF) protease inhibitor were purchased from Sigma Chemicals (Sigma-Aldrich, St. Louis, MO, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from United States Biological (Massachusetts, MA, USA). Propidium iodide (PI), Ham’s F-12 nutrient, and 10,000 U/ml penicillin/streptomycin were obtained from Invitrogen (Life Technologies, Grand Island, NY, USA). Fetal bovine serum (FBS... Andrographolide Stock Solution Preparation Andrographolide was dissolved in DMSO at a concentration of 100 mM as a stock solution and stored at −20°C. Andrographolide solution at the desired concentrations was freshly prepared by diluting from a stock solution in serum-free Ham’s F-12 media. Control experiments received only media and the same amount of DMSO. The final concentration of DMSO was adjusted to 1% for all andrographolide concentrations. Cell Culture The CCA cell lines, HuCCA-1 (Sirisinha et al., 1991) and RMCCA-1 (Rattanasinganchan et al., 2006), were kindly gifted from Prof. Stitaya Sirisinha and Assoc. Prof. Ru Tohtong, Faculty of Science, Mahidol University, respectively. KKU-100 (Sripa et al., 2005) and KKU-M213 (Uthaisar et al., 2016) CCA cell lines were purchased from the Japanese Collection of Research Bioresources Cell Bank. These cells were cultured in Ham’s F-12 medium supplement with 10% FBS and 100 U/ml of penicillin/st... See full list on Andrographolide Attenuates Migration and Invasion of CCA Cells First, we determined the effect of andrographolide on cell viauding HuCCA-1, KKU-100, KKU-M213, and RMCCA-1. The results indicated that andrographolide significantly reduced CCA cell viability in a concentration-dependent manner. The 50% inhibitory growth concentration (IC50) values of andrographolide for CCA cells were 121, 137, 134, and 134 µM for HuCCA-1, KKU-100, KKU-M213, and RMCCA-1, respectively (Figure 1A). Our results indicate that andrographolide re... Andrographolide Inhibits Claudin-1 Expression in CCA Cells To investigate the mechanism through which andrographolide inhibits CCA cell migration and invasion, we analyzed tuding vimentin, E-cadherin, claudin-1, and Snail. In this experiment, we used two cell lines, migratory KKU-M213 and non-migratory KKU-100 cells. The results showed that claudin-1 expression was substantially decreased in KKU-M213 cells upon exposure to andrographolide treatment, whereas the expression of Snail, a repressor of claudin-1, was up-re... Suppression of Claudin-1 Expression Inhibits Cell Migration of CCA Cells Transient suppression of claudin-1 in KKU-M213 cells was performed using siRNA specific to claudin-1. Cells treated with scramble RNA were used as controls. The real-time RT-PCR and Western blot analysis showed that claudin-1, compared with the controls, was successfully knocked down by its specific siRNA >8-folds (Figures 4A, B). Migration activity of KKU-M213 cells was evaluated by wound healing assay. The results revealed that the cells treated with si-claudin-1 exhibited significant reduc... See full list on Because of high mortality rate and lack of effective curative options in CCA, the need for novel therapeutic measures are urgently needed. In recent years, a number of studies have indicated that the naturally extracted compounds from herbal plants exhibit anticancer properties. Andrographolide, a major constituent from A. paniculata, has been shown to have multi-functions of anticancer activities in variety of cancer types (Lim et al., 2012 Mishra et al., 2015). The crude extract of A. paniculata displays the ability to arrest cell cycle and induce apoptosis in CCA cells (Suriyo et al., 2014). However, the effects of purified andrographolide on uding cell migration and invasion have never been described. This current study hence explored such effect together with underlying molecular mechanism using four different CCA cell lines. Andrographolide has been shown to inhibit growth in several cancer cell types such as breast cancer, colon cancer, lung... See full list on The raw data supporting the conclusions of this manuscript will be made available by the authors, without undue reservation, to any qualified researcher. See full list on PP performed experiments, analyzed the data, and wrote the manuscript SK and SO contributed to the experimental work and analyzed the data TJ conceived and designed the study, analyzed the data, wrote the manuscript, and supervised the research. All authors read and approved the final version of the manuscript. See full list on This work is supported by grants from Cerebos foundation, Medical Research Council (MRC) through UK-Thailand Research Collaborations (Newton Fund), the Thailand Research Fund (grant no. DBG5980006) and Faculty of Science, Mahidol University (TJ). PP is funded by the Science Achievement Scholarship of Thailand. The equipment was facilitated by Central Instrument Facility (CIF) at Faculty of Science, Mahidol University. See full list on The authors declare that the research was conducted in the absence of any commercial or ficial relationships that could be construed as a potential conflict of interest. See full list on
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