Histone Deacetylases and their Inhibitors as Potential Therapeutic Drugs for cholangiocarcinoma - Cell Line findings

Sriraksa, Ruethairat,Limpaiboon, Temduang Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.4

Histone deacetylation mediated by histone deacetylases (HDACs) has been reported as one of the epigenetic mechanisms associated with tumorigenesis. The poor responsiveness of anticancer drugs found with cholangiocarcinoma (CCA) leads to short survival rate. We aimed to investigate mRNA expression of HDACs class I and II, and the effect of HDAC inhibitors, suberoylanilide hydroxamic acid (SAHA) and valproic acid (VPA), in CCA in vitro. Expression of HDACs was studied in CCA cell lines (M213, M214 and KKU-100) and an immortal cholangiocyte (MMNK1) by semi-quantitative reverse transcription-PCR. SAHA and VPA, as well as a classical chemotherapeutic drug 5 -fluorouacil (5-FU) were used in this study. Cell proliferation was determined by sulforhodamine assay. $IC_{50}$ and $IC_{20}$ were then analyzed for each agent and cell line. Moreover, synergistic potentional of VPA or SAHA in combination with 5-FU at sub toxic does ($IC_{20}$) of each agent was also evaluated. Statistic difference of HDACs expression or cell proliferation in each experimental condition was analyzed by Student's t-test. The result demonstrated that HDACs were expressed in all studied cell types. Both SAHA and VPA inhibited cell proliferation in a dose-dependent manner. Interestingly, KKU-100 which was less senstitive to classical chemotheraoeutic 5-FU was highly was sensitive to HDAC inhibitors. Simultaneous combination of subtoxic doses of HDAC inhibitors and 5-FU signiicantly inhibited cell proliferation in CCA cell lines compared to single sgent treatment($P{\leq}0.01$), while sequentially combined treatments were less effective. The present study showed inhibitory effects of HDACIs on cell proliferation in CCA cell lines, with synergistic antitumor potential demonstrated by simultaneous combination of VPA or SAHA with 5-FU, suggesting a novel alternative therapeutic strategy in effective treatment of CCA.

TRAIL in Combination with Subtoxic 5-FU Effectively Inhibit Cell Proliferation and Induce Apoptosis in Cholangiocarcinoma Cells

Sriraksa, Ruethairat,Limpaiboon, Temduang Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.16

In the past decade, the incidence and mortality rates of cholangiocarcinoma (CCA) have been increasing worldwide. The relatively low responsiveness of CCA to conventional chemotherapy leads to poor overall survival. Recently, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL or Apo2L) has emerged as the most promising anti-cancer therapeutic agent since it is able to selectively induce apoptosis of tumor cells but not normal cells. In this study, we aimed to investigate the therapeutic effect of TRAIL in CCA cell lines (M213, M214 and KKU100) compared with the immortal biliary cell line, MMNK1, either alone or in combination with a subtoxic dose of 5-fluorouracil (5-FU). We found that recombinant human TRAIL (rhTRAIL) was a potential agent which significantly inhibited cell proliferation and mediated caspase activities (caspases 8, 9 and 3/7) and apoptosis of CCA cells. The combined treatment of rhTRAIL and 5-FU effectively enhanced inhibition of CCA cell growth with a smaller effect on MMNK1. Our finding suggests TRAIL to be a novel anti-cancer therapeutic agent and advantage of its combination with a conventional chemotherapeutic drug for effective treatment of CCA.

Cytotoxic Effects of Phytophenolics from Caesalpinia mimosoides Lamk on Cervical Carcinoma Cell Lines through an Apoptotic Pathway

Palasap, Adisak,Limpaiboon, Temduang,Boonsiri, Patcharee,Thapphasaraphong, Suthasinee,Daduang, Sakda,Suwannalert, Prasit,Daduang, Jureerut Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.1

Background: Extracts of Caesalpinia mimosoides Lamk has been reported to possess anticancer effects, but the active ingredients and the anti-cancer mechanisms are still unknown. Materials and Methods: The effects of a C mimosoides Lamk extract on cell proliferation and apoptosis induction in human cervical carcinoma cell lines, namely HeLa, SiHa, and C33A, as well as in normal Vero cells, were investigated. Results: Treatment with 5 active fractions (F17-F21) of C mimosoides Lamk methanol extracts inhibited cell viability in a dose- and time-dependent manner. Neutral red assays indicated that treatment with F21 significantly decreased the viability of all cervical cancer cell lines compared to F21-treated normal cells. In addition, HPLC analysis revealed that F21 contained multiple phenolic compounds, namely gallic acid, caffeine, vanillic acid, ferulic acid and resveratrol. F21 had the lowest IC50 and, therefore, a much higher cytotoxicity than F20, F17, F19, and F18 by 20-, 25-, 46- and 47- fold, respectively. Analysis of activation of the apoptosis pathway using a caspase 3/7 activity assay revealed that F21 treatment resulted in a considerable increase in caspase activation in all cancer cell lines tested. At the same concentration of F21, HeLa cells had the highest caspase activity (6.5-fold) compared to the control. Conclusion: C mimosoides Lamk may be of value as an alternative therapeutic agent, especially in combination with other compounds offering possible of synergy of action. Moreover, HPV- and non-HPV-related cervical cancer cells may differ in their responses to treatment regimens.

Serum Cathepsin B to Cystatin C Ratio as a Potential Marker for the Diagnosis of Cholangiocarcinoma

Monsouvanh, Ammala,Proungvitaya, Tanakorn,Limpaiboon, Temduang,Wongkham, Chaisiri,Wongkham, Sopit,Luvira, Vor,Proungvitaya, Siriporn Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.21

Cholangiocarcinoma (CCA) is a cancer of the bile duct epithelial cells. The highest incidence rate of CCA with a poor prognosis and poor response to chemotherapy is found in Southeast Asian countries, especially in northeastern Thailand and Lao PDR. Cathepsin B is a lysosomal cysteine protease which is regulated by cysteine proteinase inhibitors such as cystatin C. Elevation of cathepsin B levels in biological fluid has been observed in patients with inflammatory diseases and many cancers. We aimed to investigate the serum cathepsin B and cystatin C levels of CCA patients to evaluate the feasibility of using cathepsin B and cystatin C as markers for the diagnosis of CCA. Fifty-six sera from CCA patients, 17 with benign biliary diseases (BBD) and 13 from controls were collected and the cathepsin B and cystatin C levels were determined. In addition, cathepsin B expression was investigated immunohistochemically for 9 matched-pairs of cancerous and adjacent tissues of CCA patients. Serum cathepsin B, but not cystatin C, was significantly higher in CCA and BBD patient groups compared to that in the control group. Consistently, all cancerous tissues strongly expressed cathepsin B while adjacent tissues were negative in 7 out of 9 cases. In contrast, serum cystatin C levels were comparable between CCA and control groups, although serum cystatin C levels in the BBD group was higher than that in the control or CCA groups. When the serum cathepsin B to cystatin C ratio was calculated, that of the CCA group was significantly higher than that of the control group, and, although statistically not significant, the ratio of CCA group showed a trend to be higher than that of the BBD group. Thus, the cathepsin B to cystatin C ratio might be used as an alternative marker for aiding diagnosis of CCA.

Total Serum Bile Acid as a Potential Marker for the Diagnosis of Cholangiocarcinoma without Jaundice

Sombattheera, Sutthikan,Proungvitaya, Tanakorn,Limpaiboon, Temduang,Wongkham, Sopit,Wongkham, Chaisiri,Luvira, Vor,Proungvitaya, Siriporn Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.4

Diagnosis of cholangiocarcinoma (CCA) is difficult when patients do not show jaundice. The aim of this study was to examine the feasibility of using the total serum bile acid (TSBA) level as an aid for the diagnosis of CCA in patients without jaundice. For this purpose, TSBA of the following groups were measured using a Beckman Synchron CX4 clinical chemistry analyzer: 60 cases of CCA with total serum bilirubin ${\leq}2mg/dL$ (low total bilirubin group, LTB); 32 cases of CCA with total serum bilirubin >2 mg/dL (high total bilirubin group, HTB); and 115 healthy controls. Liver function parameters such as serum cholesterol, albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were also examined. The results showed that the TSBA of both LTB and HTB groups of the CCA patients were significantly higher than that of the healthy controls. Also, significant correlation was observed between TSBA and total bilirubin levels in the HTB group of CCA patients. However, no such correlation was seen in the LTB group. The cut-off value of TSBA was determined for the LTB group of CCA patients using the receiver operating characteristic curve analysis, and it was $6.05{\mu}mol/L$ with the sensitivity and specificity of 46.7% and 84.4%, respectively. In addition, the ALP level was correlated well with the TSBA level and ALP in HTB group was significantly higher than that of LTB group. Moreover, the combination of high TSBA and high ALP levels gave higher specificity up to 97.4%. TSBA might be useful for the diagnosis of CCA patients without jaundice.

Accuracy of Vesical Imaging-Reporting and Data System for muscle-invasive bladder cancer detection from multiparametric magnetic resonance imaging

Chayanon Jai-ua,Chatwadee Limpaiboon,Satit Siriboonrid,Nattapong Binsri,Sarayut Kanjanatarayon,Weerayut Wiriyabanditkul,Vittaya Jiraanankul 대한비뇨의학회 2023 Investigative and Clinical Urology Vol.64 No.6

Purpose: The Vesical Imaging-Reporting and Data System (VI-RADS) was used to distinguish the invasive nature of bladder masses before surgery. These imaging criteria can be used to carefully select patients who are candidates for repeat transurethral resection of bladder tumor (Re-TUR-BT). One-third of patients are understage at the time of Re-TUR-BT. This study aimed to evaluate the discrimination accuracy of VI-RADS between non-muscle-invasive bladder cancer and muscle-invasive bladder cancer. Materials and Methods: Patients with a bladder mass identified by cystoscopy who were assigned for TUR-BT were offered multiparametric magnetic resonance imaging (mpMRI) for VI-RADS. TUR-BT reports were compared with preoperative VI-RADS scores to evaluate the accuracy of discrimination of the muscle-invasive nature of the bladder mass. Results: A total of 58 bladder tumor lesions were included, 13 with muscle-invasive bladder cancer and 45 with non-muscle-invasive bladder cancer. Sensitivity and specificity were 92.3% and 86.7%, respectively, when a VI-RADS cutoff of 4 or more was used to define muscle-invasive bladder cancer. Positive predictive value and negative predictive value were 66.7% and 97.5%, with an accuracy of 87.9%. The area under the receiver operating characteristic curve was 0.932 (95% confidence interval, 0.874–0.989), and the empirical optimal cutpoint from the Youden method was 3. Conclusions: VI-RADS is an accurate tool for correctly differentiating muscle-invasive bladder cancer from non-muscle-invasive bladder cancer. We found a cutpoint of VI-RADS 1–3 vs. 4–5 to have the highest specificity and accuracy for the discrimination of non-muscle-invasive from muscle-invasive bladder cancer.

Gold Nanoparticles Enhance the Anticancer Activity of Gallic Acid against Cholangiocarcinoma Cell Lines

Rattanata, Narintorn,Daduang, Sakda,Wongwattanakul, Molin,Leelayuwat, Chanvit,Limpaiboon, Temduang,Lekphrom, Ratsami,Sandee, Alisa,Boonsiri, Patcharee,Chio-Srichan, Sirinart,Daduang, Jureerut Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.16

Gold nanoparticles (GNPs) were conjugated with gallic acid (GA) at various concentrations between 30 and $150{\mu}M$ and characterized using transmission electron microscopy (TEM) and UV-Vis spectroscopy (UV-VIS). The anticancer activities of the gallic acid-stabilized gold nanoparticles against well-differentiated (M213) and moderately differentiated (M214) adenocarcinomas were then determined using a neutral red assay. The GA mechanism of action was evaluated using Fourier transform infrared (FTIR) microspectroscopy. Distinctive features of the FTIR spectra between the control and GA-treated cells were confirmed by principal component analysis (PCA). The surface plasmon resonance spectra of the GNPs had a maximum absorption at 520 nm, whereas GNPs-GA shifted the maximum absorption values. In an in vitro study, the complexed GNPs-GA had an increased ability to inhibit the proliferation of cancer cells that was statistically significant (P<0.0001) in both M213 and M214 cells compared to GA alone, indicating that the anticancer activity of GA can be improved by conjugation with GNPs. Moreover, PCA revealed that exposure of the tested cells to GA resulted in significant changes in their cell membrane lipids and fatty acids, which may enhance the efficacy of this anticancer activity regarding apoptosis pathways.

Gallic Acid Enhancement of Gold Nanoparticle Anticancer Activity in Cervical Cancer Cells

Daduang, Jureerut,Palasap, Adisak,Daduang, Sakda,Boonsiri, Patcharee,Suwannalert, Prasit,Limpaiboon, Temduang Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.1

Cervical cancer (CxCa) is the most common cancer in women and a prominent cause of cancer mortality worldwide. The primary cause of CxCa is human papillomavirus (HPV). Radiation therapy and chemotherapy have been used as standard treatments, but they have undesirable side effects for patients. It was reported that gallic acid has antioxidant, antimicrobial, and anticancer activities. Gold nanoparticles are currently being used in medicine as biosensors and drug delivery agents. This study aimed to develop a drug delivery agent using gold nanoparticles conjugated with gallic acid. The study was performed in uninfected (C33A) cervical cancer cells, cervical cancer cells infected with HPV type 16 (CaSki) or 18 (HeLa), and normal Vero kidney cells. The results showed that GA inhibited the proliferation of cancer cells by inducing apoptosis. To enhance the efficacy of this anticancer activity, 15-nm spherical gold nanoparticles (GNPs) were used to deliver GA to cancer cells. The GNPs-GA complex had a reduced ability compared to unmodified GA to inhibit the growth of CxCa cells. It was interesting that high-concentration ($150{\mu}M$) GNPs-GA was not toxic to normal cells, whereas GA alone was cytotoxic. In conclusion, GNPs-GA could inhibit CxCa cell proliferation less efficiently than GA, but it was not cytotoxic to normal cells. Thus, gold nanoparticles have the potential to be used as phytochemical delivery agents for alternative cancer treatment to reduce the side effects of radiotherapy and chemotherapy.

Inhibitory Effects of Gallic Acid Isolated from Caesalpinia mimosoides Lamk on Cholangiocarcinoma Cell Lines and Foodborne Pathogenic Bacteria

Rattanata, Narintorn,Klaynongsruang, Sompong,Daduang, Sakda,Tavichakorntrakool, Ratree,Limpaiboon, Temduang,Lekphrom, Ratsami,Boonsiri, Patcharee,Daduang, Jureerut Asian Pacific Journal of Cancer Prevention 2016 Asian Pacific journal of cancer prevention Vol.17 No.3

Gallic acid was isolated from Caesalpinia mimosoides Lamk and the structure s identified based on spectroscopic analysis and comparison with authentic compound. In this study we compared the ability of natural gallic acid (nGA) and commercial gallic acid (cGA) to inhibit the proliferation of cholangiocarcinoma cell lines (M213, M214) and foodborne pathogenic bacteria (Salmonella spp. and Plesiomonas shigelloides). Both nGA and cGA had the same inhibitory effects on cell proliferation by inducing apoptosis of cholangiocarcinoma cell lines. In addition, nGA inhibited growth of foodborne pathogenic bacteria in the same manner as cGA. Our results suggest that nGA from Caesalpinia mimosoides Lamk is a potential anticancer and antibacterial compound. However, in vivo studies are needed to elucidate the specific mechanisms involved.

Contribution of RIZ1 to Regulation of Proliferation and Migration of a Liver Fluke-Related Cholangiocarcinoma Cell

Khaenam, Prasong,Niibori, Akiko,Okada, Seiji,Jearanaikoon, Patcharee,Araki, Norie,Limpaiboon, Temduang Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.8

Purpose: Retinoblastoma-interacting zinc finger gene (RIZ1) is a tumor suppressor gene which is highly inactivated by promoter hypermethylation in patients with liver fluke-related cholangiocarcinoma (CCA). Epigenetic aberration of this gene might withdraw the ability to restrain tumor cell proliferation and migration. We aimed to define the role of RIZ1 on cell proliferation and migration in CCA cell line. Materials and methods: Small interference RNA (siRNA) was used to knock down the expression of RIZ1 in a CCA-derived cell line in which cell proliferation and cell migration were performed. Results: A predominant nuclear localization of RIZ1 was observed. Reduction of RIZ1 by siRNA augmented cell proliferation and migration. Conclusion: The result suggested that RIZ1 might play a role in regulating cell proliferation and migration in CCA. Reduction of RIZ1 expression may aggravate the progression of CCA.