Peti Thuwajit

Nucleolin (NCL) is a multifunctional protein expressed in the nucleus, cytoplasm, and cell membrane. Overexpression of NCL has a controversial role as a poor prognostic marker in cancers. In this study, a meta-analysis was performed to evaluate the prognostic value of NCL in different subcellular localizations (cytoplasmic (CyNCL) and nuclear (NuNCL)) across a range of cancers. PubMed was searched for relevant publications. Data were extracted and analyzed from 12 studies involving 1221 patients with eight cancer types. The results revealed high total NCL was significantly associated with poor overall survival (OS) (HR = 2.85 (1.94, 4.91), p < 0.00001, I2 = 59%) and short disease-free survival (DFS) (HR = 3.57 (2.76, 4.62), p < 0.00001, I2 = 2%). High CyNCL was significantly associated with poor OS (HR = 4.32 (3.01, 6.19), p < 0.00001, I2 = 0%) and short DFS (HR = 3.00 (2.17, 4.15), p < 0.00001, I2 = 0%). In contrast, high NuNCL correlated with increased patient OS (HR =...

Colorectal cancer (CRC) is one of the most fatal cancers with highly invasive properties. The progression of CRC is determined by the driving force of periostin (PN) from cancer‐associated fibroblasts (CAFs) in the tumour microenvironment. This present work aims to investigate autophagy‐mediated CRC invasion via the receptor integrin (ITG) by PN. The level of PN in 410 clinical CRC tissues was found increased and was an independent poor prognosis marker (HR = 2.578, 95% CI = 1.218‐5.457, P‐value = .013) with a significant correlation with overall survival time (P‐value < .001). PN activated proliferation, migration and invasion of CRC cells, but with reduced autophagy. Interestingly, the reduction of LC3 autophagic protein corresponded to the increased ability of CRC cell migration. The siITGα5‐treated HT‐29 and siITGβ4‐treated HCT‐116 CRC cells attenuated epithelial‐to‐mesenchymal transitions (EMT)‐related genes and pAKT compared with those in siITG‐untreated cells. The reduction of pAKT by a PI3K inhibitor significantly restored autophagy in CRC cells. These evidences confirmed the effect of PN through either ITGα5β1 or ITGα6β4 and the AKT‐dependent pathway to control autophagy‐regulated cell migration. In conclusion, these results exhibited the impact of PN activation of ITGα5β1 or ITGα6β4 through pAKT in autophagy‐mediated EMT and migration in CRC cells.

BACKGROUND & AIMS Accurate differentiation between cholangiocarcinoma (CCA) and benign biliary stricture is of paramount importance. Biliary brush cytology is a simple and safe diagnostic approach that provides relatively high specificity; however, sensitivity is limited. Previous reports indicated the aberrations of DNA methylation in CCA. This study was aimed to investigate the diagnostic performance of the methylation index (MI) of HOXA1, NEUROG1 gene promoters in CCA. METHODS Patients with biliary stricture who underwent endoscopic retrograde cholangiopancreatography (ERCP) with brush cytology in Siriraj Hospital from September 2016 to December 2019 were prospectively enrolled. The MI of HOXA1 (MI_H) and MI of NEUROG1 (MI_N) were determined by quantitative methylation-specific polymerase chain reaction. The diagnostic power for CCA was tested for MI from both genes and serum CA19-9. RESULTS A total of 67 patients were included in the study; 41 patients had a final diagnosis of CCA, and 26 patients were determined to have a benign biliary stricture. The results showed that both MI_H and MI_N had higher sensitivity/accuracy (95.1%/82.3% and 90.2%/89.5%, respectively) than brush cytology (61.5%/78.1%) and CA19-9 (69.4%/77.8%). The combination of brush cytology, both methylation markers and CA19-9 increased sensitivity/accuracy to 97.4%/91.0%. Methylation markers were positive in 5 out of 6 patients with confirmed CCA whose cytology and CA19-9 were negative. CONCLUSIONS DNA methylation increased sensitivity for the diagnosis of CCA; therefore, the usage of DNA methylation is promising for diagnosis of CCA in patients with biliary strictures. A future validation study is warranted to assess its role in clinical practice.

BACKGROUND & AIMS Accurate differentiation between cholangiocarcinoma (CCA) and benign biliary stricture is of paramount importance. Biliary brush cytology is a simple and safe diagnostic approach that provides relatively high specificity; however, sensitivity is limited. Previous reports indicated the aberrations of DNA methylation in CCA. This study was aimed to investigate the diagnostic performance of the methylation index (MI) of HOXA1, NEUROG1 gene promoters in CCA. METHODS Patients with biliary stricture who underwent endoscopic retrograde cholangiopancreatography (ERCP) with brush cytology in Siriraj Hospital from September 2016 to December 2019 were prospectively enrolled. The MI of HOXA1 (MI_H) and MI of NEUROG1 (MI_N) were determined by quantitative methylation-specific polymerase chain reaction. The diagnostic power for CCA was tested for MI from both genes and serum CA19-9. RESULTS A total of 67 patients were included in the study; 41 patients had a final diagnosis of CCA, and 26 patients were determined to have a benign biliary stricture. The results showed that both MI_H and MI_N had higher sensitivity/accuracy (95.1%/82.3% and 90.2%/89.5%, respectively) than brush cytology (61.5%/78.1%) and CA19-9 (69.4%/77.8%). The combination of brush cytology, both methylation markers and CA19-9 increased sensitivity/accuracy to 97.4%/91.0%. Methylation markers were positive in 5 out of 6 patients with confirmed CCA whose cytology and CA19-9 were negative. CONCLUSIONS DNA methylation increased sensitivity for the diagnosis of CCA; therefore, the usage of DNA methylation is promising for diagnosis of CCA in patients with biliary strictures. A future validation study is warranted to assess its role in clinical practice.

Additional file 5: Figure S5. Prediction of TFATHGKHWAAP peptide structure and binding to PN. The analysis was performed by RPBS online tools ( https://bioserv.rpbs.univ-paris-diderot.fr ). (a) Prediction of peptide structure by PEPFOLD3 protein structure prediction tool. The structure is almost linear with small α-helix at N-terminal. (b) Prediction of binding between peptide (red) and PN protein (blue). The binding site is located near the active site (yellow area) of PN. The binding energy of this model was -11.89 kCal/mol.

The effects of rPAI-2 on CCA tumorigenic properties. (A) Cell proliferation was examined by WST assay in KKU-213 and KKU-055 at 24, 48, and 72 h after 10 μg/ml of rPAI-2 treatment. The 2% FBS containing media are used as negative controls. (B) Cell invasion by Transwell® invasion assay in KKU-213 and KKU-055. After incubation for 18 h with 10 μg/ml of rPAI-2, invaded cells were counted. Bars represent mean ± SD of three measurements. *P

VEGFA is a target gene of miR-15a. (A) Expression of VEGFA in miR-15a mimic transfected C096 CCFs was examined by real-time PCR. Scrambled miRNAs were used as negative control miRNA. Bars represent mean ± SD of three measurements. (B) Expression of VEGFA in miR-15a inhibitor transfected SFA3 SFs was examined by real-time PCR. Bars represent mean ± SD of three measurements. (C) Expression of VEGFA in 6 CCFs (B149, C095, C096, D005, D012, and D017), SFs (SFA3 and SFA5) and CCA cell lines (KKU-213 and KKU-055) was examined by real-time PCR. Bars represent means ± SD of three measurements. *P≤0.05 compared to control. (TIFF 78 kb)

List of candidate target genes of miR-148a. (A) Four criteria of finding the candidate target genes of miR-148a. (B) The expression levels of five predicted target genes of miR-148a including WNT10B, TGFA, WNT1, TNFRSF6B and L-selectin in CCFs and SFs. Scrambled miRNAs were used as a negative control. Bars represent mean ± SD of three measurements in one experiment. (C) Expression of WNT10B in miR-148a mimic transfected C096 CCFs was examined by real-time PCR. Bars represent mean ± SD of three measurements. *P≤0.05 compared to control. (D) Expression of WNT10B in miR-148a inhibitor transfected SFA3 SFs was examined by real-time PCR. Bars represent mean ± SD of three measurements. *P≤0.05 compared to control. (TIFF 200 kb)

The expression levels of 15 down-regulated miRNAs in in 2 CCFs (B149 and C096) and 2 normal SFs (SFA3 and SFA5). Expression of miRNAs was examined by real-time PCR. The level is normalized to U6 snRNA. Graph shows the expression of miRNAs in 2 CCFs and the average level of 2 control SFs. (TIFF 194Â kb)

Cholangiocarcinoma (CCA)is one of the worst prognosis cancers. CCA usually presents in advanced stage according to its high metastatic property. Until present, the factors involving in CCA metastasis are still unclear. Estrogen has been revealed as a stimulator of the invasiveness of CCA. Cyclooxygenase-2 (COX-2), an estrogen targeted gene, associates with cancer invasion and metastasis. This study aims to investigate the role of COX-2 in estrogen-stimulated invasion of CCA cells. KKU-100 and KKU-M213 CCA cell lines were treated with 17β-estradiol (E2), as active form of estrogen, and NS398, a selective COX-2 inhibitor, and the invasive property of cancer cells was measured as well as the expression levels of some metastatic genes. The results indicated that E2 could stimulate CCA cell invasion and this process was inhibited by NS398. In addition, in E2-stimulated CCA cells, the levels of COX-2 and CD44 were increased. These results indicate the impact of COX-2-mediated CD44 expression in E2-driven CCA invasion. The application of using COX-2 inhibitor to attenuate E2-stimulated invasion of CCA is of great interest to propose in the effective management of the disease.

Figure S1. TLR4 and MyD88 expressions of BCA cell lines. (A) Western blot analysis revealed TLR4 in all cells. Equal total protein loading was confirmed by β-ACTIN internal control. Bar graphs represent intensities of TLR4 bands quantified by ImageJ® software and normalized with that of β-ACTIN. (B) Immunocytochemistry for TLR4 in BCA cell lines. TLR4 was labeled with goat anti-human TLR4 and donkey anti-goat IgG-Alexa Fluor® 488 (green). Hoechst® 33,258 (blue) was used for nuclei staining (Scale bar = 20 μm and original magnification 400X). (C) MyD88 was detected by real-time PCR. MyD88 mRNA expression level was normalized by ACTB as an internal control. Bars represent mean ± SD of duplicate PCR reactions. (D and E) LC50 of MDA-MB231 and MDA-MB435 against PTX after 24 h incubation is shown. (TIFF 8856 kb)

Background/Aim: Pyruvate carboxylase (PC) is a major anaplerotic enzyme for generating oxaloacetate for the TCA cycle and also a key enzyme in gluconeogenesis, de novo fatty acid and amino acid synthesis in normal cells. Recent studies have identified PC overexpression in different cancers, such as breast and lung. However, the involvement of PC in colorectal cancer (CRC) is unclear. Our purpose was to investigate the PC expression levels and its correlations with potentially relevant clinical-pathological parameters in CRC. Materials and Methods: PC expression levels in tissues from 60 Thai CRC patients were investigated by immunohistochemistry while a clonogenic assay was performed for determining cell growth of HT-29 cells with PC knockdown. Results: Our results showed for the first time that high PC expression levels were significantly correlated with late stage of the cancer, perineural invasion and lymph node metastasis. The overexpression of PC was also significantly associated with poor overall and disease-free survival times of CRC patients. In addition, suppression of cancer cell growth was found in PC-deficient cell lines using CRISPR-Cas9. Conclusion: The overexpression levels of PC were correlated with CRC progression and survival times. Therefore, PC might serve as a potential clinical prognostic marker for colorectal cancer.

Cholangiocarcinoma is defined as a cancer arising from bile duct epithelial cell. The prevalence of cholangiocarcinoma is low among worldwide, however it was raised each year. Moreover, this kind of cancer is endemic in Thailand which associated with liver fluke infection, Opisthorchis viverrini. In Thailand, the prevalence of cholangiocarcinoma was about 20-time higher than in western country and may be more than 400-time in endemic area (northeastern part). Cholangiocarcinoma has poor prognosis with low 5-year survival rate due to its high metastasis rate which caused inadequate surgery while chemotherapy and radiotherapy are resisted. It is a type of chronic liver disease with altered estrogen metabolizing enzyme that could leading to the accumulation of estrogen in plasma. Estrogen was known as a promoting factor in progression of some cancer, eg. breast cancer and endometrial cancer. Our finding indicated the increasing of serum estrogen level in male cholangiocarcinoma patients and correlated with serum alkaline phosphatase and bilirubin and inverse correlated with albumin. The high level of serum estrogen could be determined as poor prognostic factor for survival of cholangiocarcinoma patients with statistically significant. In vitro studies demonstrated the effect of estrogen on cell proliferation and invasion of cholangiocarcinoma cell lines. Real time proliferative assay indicated the growth stimulated effect of estrogen on 4 cholangiocaricinoma cell lines (KKU-100, KKU-M055, KKU-M156 and KKU-M213) in dose dependent manner up to 10 nM. Results from in vitro invasion assay showed the effect of estrogen on cholangiocarcinoma cell invasion with highest effect at concentration 1 nM and this effect could be inhibited by tamoxifen. Metastasis genes expression of KKU-M213 cholangiocarcinoma cell induced by estrogen was measured by RT 2 Prolifiler TM PCR array system. Total 84 metastasis genes expression were compared with non-induced cell, 24 genes showed change in expression level more than twice which were 11 genes increased and 13 genes decreased. The pathway of estrogen induced metastasis genes in cholangiocarcinoma cells should be analyzed. The results should indicate the mechanism and control of cholangiocarcinoma invasion and metastasis, which may be introduced to the new therapeutic guideline. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1408A. doi:10.1158/1538-7445.AM2011-1408A

Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL Cholangiocarcinoma (CCA), a cancer arising from bile duct epithelial cells, is a serious health problem in Thai people. It is a progressive tumor with high metastatic potency. Recently, our group has found that periostin, an extracellular matrix protein, mainly produced from stromal activated fibroblasts of CCA has strong impact to induce cancer cell invasion. This work aims to investigate specific integrins which are utilized by periostin to drive invasion of CCA cells. The activated intracellular signal transduction pathway is also explored. Seven Thai patients-derived CCA cell lines were investigated their integrin expression patterns by real time PCR. The results indicated high level of integrin alpha6 in almost all cell lines. In addition, integrin alpha5 was also detected in not only CCA cell lines but also in immortalized non-tumorigenic biliary epithelial cell. For beta subunit integrin, the result revealed high expression levels of integrins beta1 and beta4 in CCA and non-tumorigenic cells. These results implied high level of integrins alpha5/beta1 and alpha6/beta4 in CCA cells. Flow cytometry and immunocytochemistry analysis confirmed intact molecules of these 2 integrin receptors on the membrane of M213 CCA cell line. Cell adhesion on periostin-coated surface was significantly decreased after integrin alpha5/beta1, but not alpha6/beta4, was blocked with the corresponding neutralizing antibodies. This result indicated that CCA cells preferred to use integrin alpha5/beta1 to bind with periostin. Moreover, cells treated with a neutralizing antibody against integrin alpha5/beta1 showed attenuated effect of periostin-induced invasion as similar to cells exposed to siRNA against integrin alpha5. Furthermore, we found that integrin alpha5-knocked down cells had decreased level of intracellular pAKT compared to negative control cells after activation with periostin. Taken these results together, it is likely to say that periostin activates CCA invasion through integrin alpha5/beta1 and AKT-dependent signal transduction pathway. This study suggests fibroblast-derived periostin, the stromal cell-derived substance, in promotion of CCA progression. Importantly, the obtained data highlight the potential of using the mechanism underlying stroma-driven cancer progression as an alternative target in the treatment of CCA patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 419. doi:10.1158/1538-7445.AM2011-419

Purpose Triple negative breast cancer (TNBC) is deficient in targeted treatment resulting in poor prognosis. Targeting overexpressed mesothelin (MSLN) using MSLN-specific T cells is an attractive treatment approach.Methods The immunohistochemistry of MSLN in TNBC tissues were performed. A lentiviral vector harboring granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin-4 (IL-4) and MSLN cDNAs was constructed to generate self-differentiated myeloid-derived antigen-presenting-cells reactive against tumor expressing MSLN dendritic cells (MSLN-SmartDC) for MSLN-specific T cell activation. The antigen specificity and cancer killing of activated T cells were accessed.Results The high expression of MSLN was found in 32.8% all breast cancer subtypes and 57% in TNBC. High MSLN was significantly associated with the TNBC subtype and the absence of ER, PR and HER2. MSLN-SmartDC exhibited comparable phenotype to DC generated by exogenous cytokine treatment; addition of 40s ribos...

ABSTRACT TFF1 trefoil protein is a small secretory protein expressed in a various type of carcinomas including breast cancer. TFF1 gene contained estrogen responsive element and its expression could be regulated by estrogen. In addition, estrogen has been demonstrated its ability to promote resistance to doxorubicin in estrogen receptor positive MCF-7 breast cancer cell line. Moreover, it has been reported that TFF1 can protect conjunctival cells from UV-induced apoptosis. In this study, we demonstrated the role of TFF1 in estrogen-promoted resistance to doxorubicin-induced apoptosis using MCF-7 model. Permanent knockdown of TFF1 gene in MCF-7 cell had been generated and used to test the sensitivity to doxorubicin treatment compared to parental cell in the conditions present or absent of 17b-estradiol, a potent estrogenic agent. The apoptosis cells were measured by fluorescence staining and flow cytometry method. The results showed that among the stimulation of apoptosis by doxorubicin, 17b-estradiol could suppress this process in parental MCF-7 cell but not in TFF1 knockdown MCF-7 cell. Moreover, by trypan blue staining method, it has been shown that anti-TFF1 antibody could reverse the anti-apoptotic effect of estrogen in parental MCF-7 cell and recombinant TFF1 could recover TFF1 knockdown MCF-7 cell death induced by doxorubicin. However, this process could not be inhibited by fulvestrant, an estrogen antagonist. Apoptosis protein array experiment reflected the role of the anti-oxidative enzymes catalase and heme oxygenase 1 in estrogen and TFF1 modulated apoptosis. These phenomena determine the role of TFF1 in estrogen-promoted resistance to apoptosis induced by doxorubicin in MCF-7 breast cancer cell. TFF1 gene may be a target for enhancing of sensitivity to chemotherapy in breast cancer treatment. Disclosure All authors have declared no conflicts of interest.

Signals from the tumor microenvironment (TME) have a profound influence on the maintenance and progression of cancers. Chronic inflammation and the infiltration of immune cells in breast cancer (BC) have been strongly associated with early carcinogenic events and a switch to a more immunosuppressive response. Cancer-associated fibroblasts (CAFs) are the most abundant stromal component and can modulate tumor progression according to their secretomes. The immune cells including tumor-infiltrating lymphocytes (TILs) (cytotoxic T cells (CTLs), regulatory T cells (Tregs), and helper T cell (Th)), monocyte-infiltrating cells (MICs), myeloid-derived suppressor cells (MDSCs), mast cells (MCs), and natural killer cells (NKs) play an important part in the immunological balance, fluctuating TME between protumoral and antitumoral responses. In this review article, we have summarized the impact of these immunological players together with CAF secreted substances in driving BC progression. We exp...

Background: Chemoresistance is one of the main problems in treatment of cancer. Periostin (PN) is a stromal protein which is mostly secreted from cancer associated fibroblasts in the tumor microenvironment and can promote cancer progression including cell survival, metastasis, and chemoresistance. The main objective of this study was to develop an anti-PN peptide from the bacteriophage library to overcome PN effects in breast cancer (BCA) cells. Methods: A twelve amino acids bacteriophage display library was used for biopanning against the PN active site. A selected clone was sequenced and analyzed for peptide primary structure. A peptide was synthesized and tested for the binding affinity to PN. PN effects including a proliferation, migration and a drug sensitivity test were performed using PN overexpression BCA cells or PN treatment and inhibited by an anti-PN peptide. An intracellular signaling mechanism of inhibition was studied by Western blot analysis. Lastly, PN expressions i...

Cholangiocarcinoma (CCA) has an abundance of tumor stroma which plays an important role in cancer progression via tumor-promoting signals. This study aims to explore the microRNA (miRNA) profile of CCA-associated fibroblasts (CCFs) and the roles of any identified miRNAs in CCA progression. miRNA expression profiles of CCFs and normal skin fibroblasts were compared by microarray. Identified downregulated miRNAs and their target genes were confirmed by real-time PCR. Their binding was confirmed by a luciferase reporter assay. The effects of conditioned-media (CM) of miRNA mimic- and antagonist-transfected CCFs were tested in CCA migration in wound healing assays. Finally, the levels of miRNA and their target genes were examined by real-time PCR and immunohistochemistry in clinical CCA samples. miR-15a was identified as a downregulated miRNA in CCFs. Moreover, PAI-2 was identified as a novel target gene of miR-15a. Recombinant PAI-2 promoted migration of CCA cells. Moreover, CM from mi...

Endoscopic retrograde cholangiopancreatography with brushed cytology is still the standard method for the diagnosis of extrahepatic cholangiocarcinoma in obstructive jaundice; however, the diagnostic yield is limited. To improve the diagnostic sensitivity, DNA methylation analysis is an attractive candidate, since this may constitute a stable marker in brushed specimens. Therefore, this study aims to evaluate the importance of such epigenetic markers in brushed biliary cells from patients with obstructive jaundice for the diagnosis of extrahepatic cholangiocarcinoma. The cells examined were those that were left over from brushed cytology done during routine endoscopic retrograde cholangiopancreatography of patients with extrahepatic cholangiocarcinoma. The methylation states of HOXA1, RASSF1A, P16, and NEUROG1 genes in extrahepatic cholangiocarcinoma were measured by quantitative methylation-specific polymerase chain reaction and compared between brushed biliary cells and normal gal...

Paclitaxel (PTX) is a potent anti-cancer drug commonly used for the treatment of advanced breast cancer (BCA) and melanoma. Toll-like receptor 4 (TLR4) promotes the production of pro-inflammatory cytokines associated with cancer chemoresistance. This study aims to explore the effect of TLR4 in PTX resistance in triple-negative BCA and advanced melanoma and the effect of compound A (CpdA) to attenuate this resistance. BCA and melanoma cell lines were checked for the response to PTX by cytotoxic assay. The response to PTX of TLR4-transient knockdown cells by siRNA transfection was evaluated compared to the control cells. Levels of pro-inflammatory cytokines, IL-6 and IL-8, and anti-apoptotic protein, XIAP were measured by real-time PCR whereas the secreted IL-8 was quantitated by ELISA in TLR4-transient knockdown cancer cells with or without CpdA treatment. The apoptotic cells after adding PTX alone or in combination with CpdA were detected by caspase-3/7 assay. PTX could markedly ind...

Cancer and stromal cells, which include (cancer-associated) fibroblasts, adipocytes, and immune cells, constitute a mixed cellular ecosystem that dynamically influences the behavior of each component, creating conditions that ultimately favor the emergence of malignant clones. Ovarian cancer cells release cytokines that recruit and activate stromal fibroblasts and immune cells, so perpetuating a state of inflammation in the stroma that hampers the immune response and facilitates cancer survival and propagation. Further, the stroma vasculature impacts the metabolism of the cells by providing or limiting the availability of oxygen and nutrients. Autophagy, a lysosomal catabolic process with homeostatic and prosurvival functions, influences the behavior of cancer cells, affecting a variety of processes such as the survival in metabolic harsh conditions, the invasive growth, the development of immune and chemo resistance, the maintenance of stem-like properties, and dormancy. Further, autophagy is involved in the secretion and the signaling of promigratory cytokines. Cancer-associated fibroblasts can influence the actual level of autophagy in ovarian cancer cells through the secretion of pro-inflammatory cytokines and the release of autophagy-derived metabolites and substrates. Interrupting the metabolic cross-talk between cancer cells and cancer-associated fibroblasts could be an effective therapeutic strategy to arrest the progression and prevent the relapse of ovarian cancer.

An effective serum biomarker may improve cholangiocarcinoma (CCA) management. Periostin (PN) has been demonstrated to be associated with aggressive CCA. The current study evaluated PN in blood serum for its diagnostic and prognostic potential in patients with CCA. Sera of 68 patients with CCA were collected prior to treatment, and PN levels were measured using an ELISA. Sera from 50 normal controls, 6 patients with benign liver diseases, 2 with hepatocellular carcinoma and 21 with breast cancer were analyzed. Immunohistochemistry of PN in CCA tissues was also investigated. The data were analyzed using the Mann-Whitney U test, Kaplan-Meier log rank tests, Cox proportional hazard regression models and Fisher's exact tests. The median serum PN level in patients with CCA was significantly increased compared with that in healthy controls, patients with benign liver diseases and patients with breast cancer (all P<0.05). Using an optimal threshold value of 94 ng/ml PN, the diagnosti...

Biliary obstruction is a common clinical manifestation of various conditions, including extrahepatic cholangiocarcinoma. However, a screening test for diagnosis of extrahepatic cholangiocarcinoma in patients with biliary obstruction is not yet available. According to the rationale that the biliary system plays a major role in lipid metabolism, biliary obstruction may interfere with lipid profiles in the body. Therefore, plasma lipidomics may help indicate the presence or status of disease in biliary obstruction suspected extrahepatic cholangiocarcinoma. This study aimed to use plasma lipidomics for diagnosis of extrahepatic cholangiocarcinoma in patients with biliary obstruction. Plasma from healthy volunteers, patients with benign biliary obstruction extrahepatic cholangiocarcinoma, and other related cancers were used in this study. Plasma lipids were extracted and lipidomic analysis was performed using matrix-assisted laser desorption ionization time-of-flight mass spectrometry. L...

Cholangiocarcinoma (CCA) is one of the worst prognosis cancer. The survival time of CCA patients is related to serum estrogen levels and estrogen has been found to enhance the proliferation and invasiveness of CCA cells in vitro. This has led to the suggestion that estrogen may play an important role in the progression of CCA. This study tests the relevance of the previous in vitro findings in vivo using a mouse xenograft model of CCA, and investigates possible signaling mechanisms involved. KKU-213 and KKU-139 CCA cell lines were used in the experiments, xenografted to nude mice and treated with a potent estrogenic agent, 17β-estradiol (E2), and/or with tamoxifen (TAM), an estrogen antagonist. The results demonstrated that E2 could accelerate growth of the xenograft-tumor and the effect was inhibited by TAM. PCR array screening of E2 responsive genes suggested ETV4 as a promising candidate intracellular mediator. ETV4-knockdown CCA cells were generated and these showed a diminished...

Cancer-associated fibroblasts and high mobility group box 1 (HMGB1) protein have been suggested to mediate cancer progression and chemotherapy resistance. The role of such fibroblasts in HMGB1 production in breast cancer is unclear. This study aimed to investigate the effects of cancer-associated fibroblasts on HMGB1 expression in breast cancer cells and its role in chemotherapeutic response. Breast cancer-associated fibroblasts (BCFs) and non-tumor-associated fibroblasts (NTFs) were isolated from human breast cancers or adjacent normal tissues and established as primary cultures in vitro. After confirmation of the activated status of these fibroblasts, conditioned-media (CM) were collected and applied to MDA-MB-231 human triple negative breast cancer cells. The levels of intracellular and extracellular HMGB1 were measured by real-time PCR and/or Western blot. The response of BCF-CM-pre-treated cancer cells to doxorubicin (Dox) was compared with those pre-treated with NTF-CM or cont...

To investigate the expressions of MUC1 and MUC5AC in intrahepatic cholangiocarcinoma (ICC). Association of expressions of mucins MUC1 and MUC5AC with clinical findings, metastasis, and survival of the liver fluke-associated ICC patients was determined. The expressions of MUC1 and MUC5AC mucins were examined by immunohistochemical staining in 87 cases of histologically-proven ICC. The expressions of mucins in relationship between clinicopathological significance and prognosis of the patients were evaluated. Fifty-two patients (60%) exhibited both MUC1 and MUC5AC expressions, whereas 31% expressed either MUC1 or MUC5AC, and 9% expressed neither. High MUC1 immunoreactivity displayed a significant correlation with tumor progression as reflected by vascular invasion (P<0.001), whereas high expression of MUC5AC significantly correlated with neural invasion (P = 0.022) and advanced ICC stage (P = 0.008). Patients with high expression of MUC1 had a significantly shorter survival (P = 0.0...

To investigate the mechanism of fibroblast cell proliferation stimulated by the Opisthorchis viverrini excretory/secretory (ES) product. NIH-3T3, mouse fibroblast cells were treated with O. viverrini ES product by non-contact co-cultured with the adult parasites. Total RNA from NIH-3T3 treated and untreated with O. viverrini was extracted, reverse transcribed and hybridized with the mouse 15K complementary DNA (cDNA) array. The result was analyzed by ArrayVision version 5 and GeneSpring version 5 softwares. After normalization, the ratios of gene expression of parasite treated to untreated NIH-3T3 cells of 2-and more-fold upregulated was defined as the differentially expressed genes. The expression levels of the signal transduction genes were validated by semi-quantitative SYBR-based real-time RT-PCR. Among a total of 15,000 genes/ESTs, 239 genes with established cell proliferation-related function were 2 fold-and more-up-regulated by O. viverrini ES product compared to those in cel...

Periostin (PN) is mainly produced from stromal fibroblasts in cholangiocarcinoma (CCA) and shows strong impact in cancer promotion. This work aimed to investigate the mechanism that PN uses to drive CCA invasion. It was found that ITGα5β1 and α6β4 showed high expression in non-tumorigenic biliary epithelial cells and in almost all CCA cell lines. PN had preferential binding to CCA cells via ITGα5β1 and blocking this receptor by either neutralizing antibody or siITGα5 could attenuate PN-induced invasion. After PN-ITGα5β1 binding, intracellular pAKT was upregulated whereas there was no change in pERK. Moreover, PN could not activate AKT in condition of treatment with a PI3K inhibitor. These data provide evidence that PN-activated invasion of CCA cells is through the ITGα5β1/PI3K/AKT pathway. Strategies aimed to inhibit this pathway may, thus, provide therapeutic benefits.