Value Of Cytokeratin Research Articles

A preliminary study on the detection of lymph node metastasis in cervical cancer using a quantitative RT-PCR assay.

This preliminary study aimed to assess the detection accuracy of sentinel lymph node metastasis in cervical cancer using quantitative reverse transcriptase-polymerase chain reaction. We collected cervical cancer tissues and 70 pelvic lymph node samples from patients with cervical cancer. The quantitative reverse transcriptase-polymerase chain reaction assay was performed to investigate the expression of cytokeratin 19 mRNA in cervical cancer tissues and determine the cutoff value of cytokeratin 19 mRNA between the non-metastatic and metastatic lymph nodes. The expression of cytokeratin 19 mRNA in cancer tissues was detected in all (71/71) the tumours, with a median copy number of 7.56×105/μl of RNA by quantitative reverse transcriptase-polymerase chain reaction. Sixteen lymph nodes were diagnosed as positive by pathological examination. The median copy numbers of cytokeratin 19 mRNA for positive and negative lymph nodes were 43.3×104/μl and 121.1/μl, respectively. The expression of cytokeratin 19 mRNA in pathologically positive lymph nodes was higher than that in the negative lymph nodes (P<0.0001) by quantitative reverse transcriptase-polymerase chain reaction analysis. Using a receiver operating characteristic plot, the maximum sensitivity (100%) and specificity (94.4%) were obtained when the cutoff value was set at 1169 copies/μl. After setting the cutoff value at 1169 copies/μl, a quantitative reverse transcriptase-polymerase chain reaction assay using cytokeratin 19 mRNA showed high accuracy in detecting lymph node metastasis in cervical cancer. We believe that the quantitative reverse transcriptase-polymerase chain reaction assay using cytokeratin 19 mRNA may be acceptable for lymph node metastasis detection in patients with cervical cancer.

Predictive Value of Serum Cytokeratin 19 Level for the Feasibility of Conserving Ovaries in Endometrial Cancer.

Objective: To determine the predictive value of cytokeratin 19 (CK19) for evaluating the safety of ovarian preservation in patients with endometrial cancer (EC).Methods: Five hundred and seventeen EC patients hospitalized from November 2010 to June 2016 were reviewed retrospectively. Pre-operative tumor biomarkers including CA125, HE4, CK19, and CA19-9 were obtained. Predictive biomarkers associated with ovarian metastasis were selected using univariate and multivariate Logistic regression. The cut-off values were determined by receiver operating characteristic (ROC) curves. Kaplan-Meier method and Cox multivariate regression model was used to perform survival analysis.Results: Among clinical parameters and biomarkers included, age > 65, type II EC, CA125 ≥ 35 u/ml, CK19 > 3.3 ng/ml, and myometrial invasion ≥ 50% depth appeared as significant predictors of the risk of ovarian involvement in univariable logistic analysis. In multivariable analysis, CK19 > 3.3 ng/ml (OR = 11.541, 95%CI: 1.968–67.668, P = 0.007) and Type II EC (OR = 8.336, 95%CI: 1.456–47.722, P = 0.017) were independent risk predictors of ovarian metastasis in pre-menopausal women. In pre-menopausal women with Type I EC (n = 142), CK19 level could satisfactorily predict the risk of ovarian metastasis (AUC = 0.860, 95%CI: 0.792–0.912, P < 0.001), and when the cut-off point was set as 2.45 ng/ml, the negative predictive value and negative likelihood ratio were 99% and 0.19, with the maximum Youden index of 0.598.Conclusions: The present study advocates the necessity of incorporating serum CK19 measurement into the pre-operative evaluation of EC, especially as extension of current standard approach with ovarian preservation counseling.

Screening diagnostic biomarkers of OSCC via an LCM-based proteomic approach.

The current standard for the diagnosis of oral squamous cell carcinoma (OSCC) is based on the histologic examination of hematoxylin and eosin-stained sections; however, the discrimination among normal tissue, pre‑cancerous lesions and cancerous lesions can be difficult. The aim of the present study was to identify proteins with diagnostic significance in differentiating or predicting oral mucosal carcinogenesis. Proteomic profiling based on the laser capture microdissection of formalin-fixed, paraffin-embedded samples was performed, followed by liquid chromatography-tandem mass spectrometry (LC/MS) analysis. Immunohistochemistry (IHC) was used to evaluate the results. IHC of cytokeratins (CKs) was performed in neck dissection treatment cases. The accuracy rate and 95% confidence intervals (CIs) were used to evaluate the value of CKs as biomarkers of OSCC. A lymph node metastasis mouse model was used to validate the selected biomarkers. Among the proteins identified using LC/MS, several CKs exhibited significant differential expression patterns between the cancerous and para-cancerous tissues. The IHC results showed that negative staining of CK4 and CK10/13 distinguished cancerous from para-cancerous tissues with an accuracy of 90% (95% CI, 0.68-0.99) and 75% (95% CI, 0.51-0.91), respectively. Furthermore, the positive staining of CK14 and CK17 clearly distinguished cancerous from para-cancerous lesions with an accuracy of 100% (95% CI, 83-100%) and 90% (95% CI, 0.68-0.99), respectively. There was also CK14-positive staining in micro-metastases of lymph nodes in the clinical samples and in an animal model.

Quantitative assesment of coexpression of cytokeratins and mesenchimal cells marker vimentin in serous ovarian cancer

Background. Epithelial-mesenchymal transition (EMT) is a factor related to metastatic potential of tumor cells. The most distinguishing feature of this transformation is expression of protein vimentin, which is not common for epithelial cells. Data of EMT level and clinical significance of the marker is ambiguous in prognosis of tumor different localization including ovarian cancer. Objective is characterization of EMT in ovarian cancer tissue based on quantitative analysis of co-expression level of epithelial cytokeratins and mesenchymal cells marker vimentin. Materials and methods. A quantitative assessment of co-expression level of cytokeratins and vimentin was performed by double immunofluorescence staining method (58 surgery specimens of ovarian cancer stage III), associated with flow cytometry. Results. Double immunofluorescence staining method, developed and used in the current study, was used for quantitative assessment of EMT level based on value of cytokeratin and vimentin co-expression in epithelial cells. Epithelial cells co-expressed these markers, were detected in 100 % tumor investigated with individual value differences from 10 to 86 %. Average co-expression level of cytokeratins and vimentin in subgroups with high and low level value related to median 42 % significantly varied and were 28,5 ± 7,5 and 56,3 ± 11,8 % (p = 0,02) respectively. Conclusions Serous ovarian cancer is molecular heterogeneous group with principal differences in level of EMT tumor phenotype between various patients. In half of the cases the disease is characterized by high metastatic potential of tumor cells. Co-expression of cyto-keratins and vimentin in all the tumors investigated is evidenced clinical significance of this molecular characteristic in ovarian cancer pathogenesis.

Expression of cytokeratin 17 mRNA in oral squamous cell carcinoma cells obtained by brush biopsy: preliminary results

The aim of this study was to determine the detection of cytokeratin (CK) mRNA in oral squamous cell carcinoma (OSCC) cells and to evaluate the CK relevance for OSCC diagnosis in a brush biopsy test. Fifty-two pairs of OSCC cells and normal oral mucosal cells were obtained by brush biopsy from OSCC patients. mRNA was extracted from cell pellets for real-time quantitative reverse transcriptase polymerase chain reaction (RT-qPCR). The over-expression levels of CK 17, CK 19 and CK 20 mRNA in OSCC cells were examined by SYBR green real-time RT-qPCR. Compared to normal mucosal cells, the over-expression of CK 17 mRNA was detectable in 40 OSCC cells (76.9%), that of CK 19 mRNA in 19 (36.5%), while that of CK 20 mRNA was not detectable. Compared with CK 19, the mean value of CK 17 mRNA expression level was significantly higher in all 52 patients (P < 0.02). Moreover, the value of CK 17 was significantly higher in T1 and T2 OSCC patients (P < 0.03, respectively), in patients without metastases of neck lymph nodes (P < 0.04), in stage I and stage II patients (P < 0.03 and P < 0.05, respectively) and in well differentiated OSCC patients (P < 0.05). Brush biopsy properly serves for detection of CK mRNA using real-time RT-qPCR. This preliminary study demonstrates the CK 17 possibility for application; however, pivotal studies are encouraged to confirm CK 17 as a diagnostic marker of OSCC in a brush biopsy test.

The Diagnostic Value of TTF-1, CK 5/6, and p63 Immunostaining in Classification of Lung Carcinomas

This study was aimed to evaluate the utility of a panel of antibodies, consisting of thyroid transcription factor-1 (TTF-1), p63, and cytokeratins (CK) 5/6 for distinguishing between small cell lung carcinoma (SCLC) and nonsmall cell lung carcinoma, as well as for identifying glandular or squamous differentiation in small tissues obtained by bronchoscopy. Bronchoscopic biopsies of 77 lung carcinoma cases with easily recognizable morphologic features were included in this study. All the cases were immunohistochemically stained for p63, CK5/6 [indicators of squamous cell carcinoma (SCC)] and TTF-1 [indicator of SCLC and adenocarcinoma (AC)]. Although, 28 SCLC displayed TTF-1 positive, p63 negative immunoprofile, most of the SCC (32/39) had the opposite immunoprofile. All of the 10 ACs were negative for p63 and most of them (8/10) were negative for CK5/6. p63 and CK 5/6 seem to be useful for differentiating AC and SCLC from SCC with 100% specificity and 82% sensitivity, 89% specificity and 79% sensitivity, respectively. It seems that to achieve histologic typing of lung cancer as accurate as possible, TTF-1 in combination with p63 and CK 5/6 might be useful components of immunohistochemical analysis of poorly differentiated lung carcinomas in biopsy tissues.

Cytokeratin 17 mRNA expression has potential for diagnostic marker of oral squamous cell carcinoma

Determination of marker for identification of oral squamous cell carcinoma (OSCC) is important for early diagnosis and individual therapy. Cytokeratins (CKs) like CK 19 and CK 20 are known to be useful diagnostic and prognostic markers for solid tumors. The aim of this study was to evaluate the relevance of further CKs for diagnosis of OSCC. In 10 OSCC and 5 normal mucosal samples, the expression patterns of 31 CK genes were examined by cDNA microarray in order to identify CKs with most pronounced over-expression. The results were verified for CK 17, CK 19, and CK 20 in addition to 46 OSCC samples by relative quantification (RQ) using SYBR green real-time reverse transcriptase polymerase chain reaction (RT qPCR). A correlation of the CK expressions with the tumor classification was carried out. cDNA microarray analyses showed that out of all CKs, CK 17 was up-regulated strongest in OSCC compared to normal samples, and over-expression was most significantly associated with diagnosis (P = 0.002). Expression rates of CK 19 and CK 20 were not significantly different between OSCC samples and normal samples. In 56 samples analyzed by real-time RT qPCR, CK 17 was over-expressed in 53 (94.6%), CK 19 in 18 (32.1%), and CK 20 in 7 (12.5%). The over-expression of CK 17 was significantly associated with metastases of neck lymph nodes (P < 0.05). CK 19 was significantly over-expressed in T3 and T4 OSCC, in stage III and IV patients (P < 0.05), and in poorly differentiated OSCC (P < 0.03). The over-expression of CK 20 was significantly associated with metastases of neck lymph nodes (P < 0.03). Determined by RQ, the mean value of CK 17 over-expression was significantly higher than that of the other CKs (P < 0.01), and was significantly associated with T1 and T2 OSCC (P < 0.03) and with stage I and II patients (P < 0.01). CK 19 might be linked to the clinical progression and differentiation of OSCC, and CK 20 could be associated with metastases of neck lymph nodes in OSCC. Due to the significant up-regulation and the strong over-expression, CK 17 might be the most suitable marker for diagnosis of OSCC out of the CK-family.

Prognostic Value of Cytokeratin 19 Fragment (CYFRA 21–1) and Cytokeratin-Positive Cells in Bone Marrow Samples of Breast Cancer Patients

The aim of this study was to investigate the relationship between the detection of micrometastatic cells by immunocytochemistry (ICC) with an anticytokeratin antibody and cytokeratin fragment (CYFRA 21–1) expression detected by an immunofluorescent assay in bone marrow of breast cancer patients. Micrometastatic CK+ cells were screened with a pancytokeratin antibody A45 B/B3 from bone marrow aspiration samples of 102 breast cancer patients (65 primary tumors, 10 local recurrences and 27 distant metastases). CYFRA 21-1 levels were assessed in bone marrow supernatant of these patients before collection of the mononucleated interface cells on a Ficoll-Hypaque density gradient and in 20 control patients. CYFRA 21-1 and CK+ cell detection by ICC were both correlated with clinical stage. CYFRA 21-1 was significantly elevated in patients with micrometastatic disease detected by ICC: 4.77 ng/mL (± 10.87 SD) versus 1.00 ng/mL (± 1.36 SD) in patients with negative ICC (p=0.01). In univariate analysis, a CYFRA 21-1 value ≥1 ng/mL and the presence of CK+ cells were associated with a poorer survival for patients with stage I to III breast cancer (n=65). On multivariate analysis, only pathological nodal status and presence of CK+ cells in bone marrow were independent prognostic factors for overall survival. In conclusion, in this series CYFRA 21-1 was correlated with detection of CK+ cells by ICC in bone marrow, but cannot replace ICC. The presence of CK+ cells in bone marrow remains a strong independent prognostic factor in primary breast cancer.

Teleangiectatic metastatic breast carcinoma in face and scalp mimicking cutaneous angiosarcoma

Teleangiectatic metastatic breast carcinoma in face and scalp mimicking cutaneous angiosarcoma

Reply: Value of cytokeratin 20 stain in Merkel-cell carcinoma

Reply: Value of cytokeratin 20 stain in Merkel-cell carcinoma