Sakkarn Sangkhamanon

Xanthohumol inhibits STAT3 activation pathway leading to growth suppression and apoptosis induction in human cholangiocarcinoma cells

STAT3 plays a significant role in the development of cholangiocarcinoma (CCA) associated with the liver fluke (Opisthorchis viverrini; Ov). Xanthohumol (XN), a prenylated flavonoid extracted from hops, has known anticancer activity and could potentially target STAT3. The present study determined the effect of XN on STAT3, as well as ascertained its usefulness against CCA. The CCA cell proliferation at 20 µM and 50 µM of XN was shown to inhibited, while 20 µM partially inhibited IL-6-induced STAT3 activation. At 50 µM, the inhibition was complete. The reduction in STAT3 activity at 20 and 50 µM was associated with a significant reduction of CCA cell growth and apoptosis. We also found that the administration of 50 µM XN orally in drinking water to nude mice inoculated with CCA led to a reduction in tumor growth in comparison with controls. In addition, apoptosis of cancer cells increased although there was no visible toxicity. The present study shows that XN can inhibit STAT3 activation both in vivo and in vitro due to suppression of the Akt-NFκB signaling pathway. XN should be considered as a possible therapeutic agent against CCA.

Inhibition of l-type amino acid transporter 1 activity as a new therapeutic target for cholangiocarcinoma treatment

Unlike normal cells, cancer cells undergo unlimited growth and multiplication, causing them to require massive amounts of amino acid to support their continuous metabolism. Among the amino acid transporters expressed on the plasma membrane, l-type amino acid transporter-1, a Na+-independent neutral amino acid transporter, is highly expressed in many types of human cancer including cholangiocarcinoma. Our previous study reported that l-type amino acid transporter-1 and its co-functional protein CD98 were highly expressed and implicated in cholangiocarcinoma progression and carcinogenesis. Therefore, this study determined the effect of JPH203, a selective inhibitor of l-type amino acid transporter-1 activity, on cholangiocarcinoma cell inhibition both in vitro and in vivo. JPH203 dramatically suppressed [14C]l-leucine uptake as well as cell growth in cholangiocarcinoma cell lines along with altering the expression of l-type amino acid transporter-1 and CD98 in response to amino acid depletion. We also demonstrated that JPH203 induced both G2/M and G0/G1 cell cycle arrest, as well as reduced the S phase accompanied by altered expression of the proteins in cell cycle progression: cyclin D1, CDK4, and CDK6. There was also cell cycle arrest of the related proteins, P21 and P27, in KKU-055 and KKU-213 cholangiocarcinoma cells. Apoptosis induction, detected by an increase in trypan blue–stained cells along with a cleaved caspase-3/caspase-3 ratio, occurred in JPH203-treated cholangiocarcinoma cells at the highest concentration tested (100 µM). As expected, daily intravenous administration of JPH203 (12.5 and 25 mg/kg) significantly inhibited tumor growth in KKU-213 cholangiocarcinoma cell xenografts in the nude mice model in a dose-dependent manner with no statistically significant change in the animal’s body weight and with no differences in the histology and appearance of the internal organs compared with the control group. Our study demonstrates that suppression of l-type amino acid transporter-1 activity using JPH203 might be used as a new therapeutic strategy for cholangiocarcinoma treatment.

Inhibitory effect of NVP‑BKM120 on cholangiocarcinoma cell growth

Abnormal activation of the phosphatidylinositol 3-kinase (PI3K) pathway has been demonstrated in certain types of cancer, including cholangiocarcinoma (CCA). This pathway may therefore be a promising target for CCA treatment. The present study assessed the inhibitory effect of NVP-BKM120, a pan-class I PI3K inhibitor, on CCA cell growth. This inhibitory effect was determined using CCA cell lines and in CCA-inoculated mice. The result from sulforhodamine B (SRB) assay demonstrated that NVP-BKM120 treatment inhibited CCA cell growth in a dose-dependent manner, even at the lowest tested concentration. The in vivo study revealed that oral administration of NVP-BKM120 (10 or 30 mg/kg) to CCA-inoculated nude mice led to a reduction in tumor growth when compared with controls, which was indicated by an immunohistochemical assay for Ki67 expression. In addition, the result from TUNEL assay demonstrated that NVP-BKM120 induced cancer cell death without any signs of toxicity, which indicated by the body weight of mice (data not shown). Western blot analysis demonstrated that NVP-BKM120 inhibited CCA cell growth by suppressing RAC serine/threonine protein kinase/mechanistic target of rapamycin activation and inhibiting the phosphorylation of phosphatase and tensin homolog, which is the inactivation form of the negative regulator of this pathway. Therefore, the results of the present study indicated that NVP-BKM120 should be considered as a therapeutic agent against CCA that could be used to improve treatment.

Bladder Malakoplakia in Systemic Sclerosis Patient: A Case Report and Review Literature

Abstract Malakoplakia, an anecdotal reactive granulomatous lesion, is a rare pathologic entity but relatively more common in genitourinary tracts. Here we report a case of malakoplakia in the urinary bladder in systemic sclerosis. The patient was a 66-year-old female treated with long-term corticosteroid and cyclophosphamide. She presented with gross hematuria, and cystoscopy and biopsy revealed malakoplakia. There was no tumor and the stains for infectious organism were all negative. To the best of our knowledge, this is the first case report of malakoplakia in a systemic sclerosis patient.

Programmed Death-Ligand 1 (PD-L1) Expression Associated with a High Neutrophil/Lymphocyte Ratio in Cholangiocarcinoma

Background: Effective treatments for cholangiocarcinoma (CCA) are still lacking. There are promising results of checkpoint inhibitor programmed cell death ligand-1 (PD-L1) activities in early phase trials. This study aimed to investigate the expression of PD-L1 and its relation to possible treatments for CCA. Methods: Formalin-fixed paraffin-embedded tumor samples from 46 patients with cholangiocarcinoma were retrieved. PD-L1 expression was evaluated by immunohistochemistry using anti-PD-L1 antibody, clone 5H1. A PD-L1 positive response on tumor cells was defined as >1% of tumor cell membranes stained. The association between PD-L1, clinico-pathological characteristics was analyzed using Fisher’s exact test, and survival analysis was done with the Cox regression model. Results: Out of 46 samples, 32 (70%) had positive PD-L1 expression in tumor cell membranes. The median level of PD-L1 expression was 1.75% (0-34.7). PD-L1 expression was significantly associated with stage IV disease (OR 3.98, p=0.046) and a high neutrophil/lymphocyte ratio (OR 5.36, p=0.018). PD-L1 positivity was associated with worse overall survival compared with those with a PD-L1 negative tumor but did not reach a level of significance (7.2 vs. 7.9 months, p=0.32). Conclusion: PD-L1 is widely expressed in CCA but was not predictive for overall survival. PD-L1 positivity was (7.2 and 7.9 months, p=0.32). Significantly associated with stage IV disease and a high neutrophil/lymphocyte ratio.

Kimura disease masquerading as soft tissue tumor of the elbow

Kimura disease is a rare chronic inflammatory disorder of unknown etiology, characterized as painless subcutaneous mass which is rarely found outside head and neck region. We report an unusual presentation of Kimura disease of a 50-year-old man who presented with a progressive large non-tenderness right elbow mass for one year. Physical examination revealed a 10 cm soft tissue mass at the medial side of his right elbow. The Laboratory investigation showed peripheral eosinophilia. The clinical diagnosis was a soft tissue tumor. It also included the differential diagnosis of a parasitic infection. Surgical excision had been performed and the histology demonstrated a nodular lymphoid hyperplasia with prominent germinal centers, a large infiltration of eosinophils with a foci of eosinophilic micro-abscess and vascular proliferation which compatible with Kimura disease. Following surgery the patient visited again with the following symptoms, a left post auricular mass with multiple cervical lymph node enlargements. The excisional biopsy revealed the same histology as the elbow.

Predictive relevance of tumor-infiltrating lymphocytes in breast cancer

&NA; Tumor-infiltrating lymphocytes (TILs) are considered to be a manifestation of the host anti-tumor response. We aim to evaluate the prognostic significance of the immunophenotype, density and distribution of TILs in breast cancer samples. This was a retrospective study, using paraffin-embedded samples obtained from 43 breast cancer patients, treated at Mahasarakham Hospital between 2012 and 2013. TILs and prognostic markers were evaluated by immunohistochemical staining of tissue microarray cores, employing monoclonal antibodies to lymphocyte markers (i.e., CD3, CD4, CD8 and CD20) and markers for breast cancer (i.e., ER, PR, HER2 and Ki-67). TILs were categorized into: 1) intratumoral lymphocytes (ITLs) when found within tumor cell nests), and 2) peritumoral lymphocytes (PTLs) when infiltrating into the stroma, adjacent to the invasive front of tumor. Results: A positive correlation was found between high density CD8+ PTLs and breast carcinoma without lymph node metastasis (p = 0.044), and an inverse correlation was found between the density of CD4+ PTLs and estrogen receptor expression (p = 0.027). The density of CD8+ PTLs as well as the number of CD20+ ITLs were independently, positively correlated with tumor size.