Does Neutrophil to Lymphocyte Ratio Have a Role in Identifying Cytokeratin 19-Expressing Hepatocellular Carcinoma?

Background: Cytokeratin 19-positive (CK19(+)) hepatocellular carcinomas (HCC) are generally associated with poor prognosis after hepatectomy. It is typically detected from postoperative immunochemistry. We have analyzed several clinically available biomarkers, in particular, neutrophil to lymphocyte ratio (NLR) and aim to develop a panel of biomarkers in identifying CK19 expression in (HCC) preoperatively. Methods: We retrospectively reviewed 36 HCC patients who underwent liver resections during January 2017 to March 2018 in Chang Gung Memorial Hospital. Patients were grouped based on the status of CK19 expression and their baseline characteristics, perioperative and oncologic outcomes were compared. Novel biomarkers including NLR, alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA) and uric acid were analyzed and correlated with CK19 expression. Results: NLR is highly associated with CK19 expression. NLR alone gave an AUROC of 0.728 (p-value = 0.043), higher than AFP, CEA or tumor size alone. NLR when combined with AFP, CEA and uric acid, gave an AUROC as high as 0.933 (p-value = 0.004). Conclusion: The current study demonstrated the predictive capability of NLR in combination with AFP, CEA and uric acid for CK19 expression in HCC patients preoperatively. Further prospective, large-scale studies are warranted to validate our findings.

Engrailed Homeobox 1 and Cytokeratin 19 Are Independent Diagnostic Markers of Eccrine Porocarcinoma and Distinguish It From Squamous Cell Carcinoma

Objectives The diagnostic utility of En1 in the histopathologic differentiation of eccrine porocarcinoma (EPC) from invasive squamous cell carcinoma (SCC) was investigated. Methods Expression of En1 and CK19 in 16 cases of EPC was immunohistochemically examined and compared with that in 32 cases of SCC. Results In all 16 EPCs, En1 was expressed in 3% to 100% of tumor cells. In 20 of the 32 SCCs, En1 was expressed in 3% to 90% of tumor cells. A total of 13 of the 16 EPCs and five of the 32 SCCs were judged as En1 positive, with a cutoff value of 25%. In addition, 11 of the 16 EPCs and four of the 32 SCCs were CK19 positive. The frequencies of En1- and CK19-positive cases were significantly higher in EPCs than in SCCs. In a logistic regression analysis for predicting EPC, En1 and CK19 were independent markers. When expression patterns of En1 and CK19 were combined, none of the 32 SCCs was both positive. In contrast, 15 of the 16 EPCs were positive for either En1 or CK19. Conclusions A combination of En1 and CK19 expression can improve the accuracy of histologic diagnosis of EPC.

Direct effects of transforming growth factor-β1 signaling on the differentiation fate of fetal hepatic progenitor cells

Aim: To investigate direct roles of TGF-β1 signaling in the differentiation process of fetal hepatic progenitor cells (HPCs). Materials & methods: Exogenous TGF-β1 and SB431542 were added into fetal HPCs. Then, SB431542 was intraperitoneally injected into pregnant mice for 8 days. Results: Fetal HPCs treated with TGF-β1 differentiated into cholangiocytes. However, hepatocyte marker was highly expressed after inhibiting TGF-β1 signaling. In vivo, hematopoietic cells were gradually replaced with liver cells and TGF-β1 expression was evidently decreased as fetal liver developed. Inhibition of TGF-β1 signaling caused increase of ALB+ cells, but CK19 expression was more obvious in control mice livers. Conclusion: TGF-β1 signaling may play decisive roles in fetal HPCs differentiation into functional hepatocytes or cholangiocytes.

Global DNA 5hmC and CK19 with 5hmC positives cell contents represent a promising biomarker for predicting prognosis in small hepatocellular carcinoma

Background: 5-Hydroxymethylcytosine (5hmC) exists dynamically and exhibits various regulatory functions. It’s possibly associated with tumor occurrence and malignant transformation. Nevertheless, the part of 5hmC in small hepatocellular carcinoma (SHCC) is still elusive. The study was directed toward characterizing 5hmC content in SHCC and assessing if global genomic 5hmC content was able to serve as a promising factor for predicting clinical results.Methods: The expression contens of 5mC, 5hmC and 5fC were assesed using ultra-high performance liquid chromatography tandem mass-spectrometry(UHPLC-MS/MS). 5hmC contens and CK19 expression were measured by immunohistochemistry(IHC) .Results: The findings in the study displayed that global genomic 5mC, 5hmC, and 5fC contents in SHCC specimens were globally reduced relative to para-tumor tissues (P<0.001), Lymph node metastasis may be found in the small HCC, the non-metastasis group exhibited higher 5mC and 5hmc contents relative to those in metastasis group (P<0.001). HBV DNA was relevant to the decrease in 5mC, 5hmC and 5fC, as evidenced by the measurements in cell lines carrying or not carrying HBV DNA. Moreover, the correlation analysis showed the negative correlation of the content of 5hmC with CK19 expression in SHCC. The decreases of both 5hmC and CK19 with 5hmC positives cell contents in genomic DNA were related to SHCC patients’ unfavorable prognosis. Compared with para-tumor tissues, the 5hmC content in SHCC specimems was dramatically reduced.Conclusions: 5hmC and CK19 with 5hmC positives cell contents in genomic DNA possibly serve as a promising biomarker for predicting prognosis in small hepatocellular carcinoma.

MIST1, an Inductive Signal for Salivary Amylase in Mesenchymal Stem Cells

Sjögren’s syndrome (SjS) is an autoimmune disease that destroys the salivary glands and results in severe dry mouth. Mesenchymal stem cell (MSC) transplantation has been recently proposed as a promising therapy for restoring cells in multiple degenerative diseases. We have recently utilized advanced proteomics biochemical assays to identify the key molecules involved in the mesenchymal-epithelial transition (MET) of co-cultured mouse bone-marrow-derived MSCs mMSCs with primary salivary gland cells. Among the multiple transcription factors (TFs) that were differentially expressed, two major TFs were selected: muscle, intestine, and stomach expression-1 (MIST1) and transcription factor E2a (TCF3). These factors were assessed in the current study for their ability to drive the expression of acinar cell marker, alpha-salivary amylase 1 (AMY1), and ductal cell marker, cytokeratin19 (CK19), in vitro. Overexpression of MIST1-induced AMY1 expression while it had little effect on CK19 expression. In contrast, TCF3 induced neither of those cellular markers. Furthermore, we have identified that mMSCs express muscarinic-type 3 receptor (M3R) mainly in the cytoplasm and aquaporin 5 (AQP5) in the nucleus. While MIST1 did not alter M3R levels in mMSCs, a TCF3 overexpression downregulated M3R expressions in mMSCs. The mechanisms for such differential regulation of glandular markers by these TFs warrant further investigation.

Diagnostic Value of TROP-2 and CK19 Expression in Papillary Thyroid Carcinoma in Both Surgical and Cytological Specimens

Papillary thyroid carcinoma (PTC) represents the most common primary malignant thyroid tumor and its diagnosis is dependent on the presence of classic nuclear features that are sometimes seen in some non-neoplastic and benign lesions. Several immunohistochemical markers are used individually or in combination to help in differentiation of PTC from mimickers. The aim of the current study was to assess the diagnostic value of TROP-2 and cytokeratin 19 (CK19) expression in differentiating PTC from other mimickers both singly and in combination. The current study was carried out on 77 surgical specimens (56 PTC and 21 non-neoplastic cases) and 12 cytological specimens (4 THY2, 6 THY4, and 2 THY5). TROP-2 was negative in 81% of non-neoplastic surgical specimens and in 100% of THY2 cytological specimens while it was positive in 71.4% of PTC surgical specimens and 100% of THY4/THY5 cytological specimens. Sensitivity and specificity of TROP-2 positive expression for diagnosis of PTC in surgical specimens reached 71% and 81%, respectively, while it reached 100% for both in cytological specimens. Cytokeratin 19 showed positive expression in 85.7% of non-neoplastic surgical specimens and in 92.9% of PTC surgical specimens. Cytokeratin 19 showed negative expression in 75% of Thy2 cases while it was positive in all studied Thy4 and Thy5 cases. Sensitivity and specificity of CK19 total estimated score for diagnosis of PTC in surgical specimens were 78.6% and 66.7%, respectively, while it reached 100% and 75% in cytological specimens. Positive TROP-2 and CK19 expression in PTC were associated with lymph node metastasis. TROP-2 is a specific rather than sensitive marker while CK19 is a sensitive rather than specific marker in differentiating PTC from other mimickers in surgical specimens. The diagnostic validity of both markers was superior in diagnosis of classic PTC compared with follicular variant PTC. TROP-2 is superior to CK19 in diagnosis of PTC in cytological specimens. Both TROP-2 and CK19 could be used preoperatively in adjunct to hematoxylin and eosin for more confident diagnosis of thyroid cytology and along with radiology as predictors of lymph node metastasis.

Expression of epithelial marker CK19 in mice with mammary adenocarcinoma afterexposure to fito-anti-estrogen secoisolariciresinol

Introduction.Phyto-anti-estrogen secoisolariciresinol (SECO) has similar effectiveness to that of Tamoxifen (TAM), a member of selection estrogen receptor modulators, in estrogen-positive models of murine mammary adenocarcinoma and human breast cancer in vivo. Due to its antagonistic and agonistic functions Tamoxifen may enhance risk of development of uterine adenocarcinoma while providing effective prophylactics of breast cancer metastases. SECO effect on proliferative activity of circulating or disseminating tumor cells (occult metastases) is still unclear. We used epithelial cell marker – intracellular cytokeratin 19 (CK19) to study SECO function in terms of possible metastatic process, since prognostic significance of CK19 is well established for identifying occult metastases and breast cancer dissemination.Objective.To evaluate risk of tumor cell dissemination in mice with transplanted mammary adenocarcinoma after exposure to SECO and TAM.Materials and methods.Mice BDF 1 [C57Bl6j × DBA2] bearing Са755 of the 3rd passage were used for the experiments. CK19 expression was evaluated 24 hours after 10-day course of SECO in the effective single doses of 250 mg/kg or ТАM 50 mg/kg. Flow cytometry, immunofluorescence with light and luminescence microscopy were performed to evaluate CK19 expression.Results.Parameter GMFI ± SD (geometric mean fluorescence intensity ± SD) for CK19 expression in SECO and TAM groups in blood accounted for 28.87 ± 13.70 and 28.02 ± 9.50 and in control tumor growth (CTG) group GMFI ± SD was 31.94 ± 5.02; while in bone marrow it was 30.14 ± 2.33, 39.07 ± 2.30 and 32.48 ± 3.75, respectively.Conclusion.The results of the study showed similar expression of epithelial intracellular marker CK19 in blood in the studied groups of mice bearing mammary adenocarcinoma Ca755 sensitive to SECO and TAM exposure. GMFI for CK19 expression in bone marrow was lower in SECO group than in TAM (р = 0.0003). The data obtained in the murine model demonstrated no enhanced risk of tumor cell dissemination while performing treatment by phyto-anti-estrogen SECO in therapeutic regimen.