[Influence of EphA2 siRNA transfection on the biological behavior of human ovarian carcinoma cell].

OBJECTIVE: To explore EphA2 siRNA transfection and its influence on the biological behavior of human ovarian carcinoma cells. METHODS: One pairs of siRNA was synthesized and transfected into the human ovarian carcinoma cell line SKOV3. Observation of the transfection efficiency through the fluorescence inverted microscope was followed by the evaluation of expression of EphA2 protein using Western blot. The effects of EphA2 siRNA on proliferation of SKOV3 cells were observed by drawing cell growth curves and measuring cloning efficiency, and the adhesion and invasion of SKOV3 were detected by cell adhesion assay and Matrigel-boyden chamber method respectively. RESULTS: After successful transfection, a large number of fluorescent particles were observed under the fluorescence inverted microscope. The expression of EphA2 protein was obviously suppressed by EphA2 siRNA (P < 0.05). The cell proliferation, adhesiveness and invasiveness were significantly inhibited (P < 0.05). However, no changes in colony-forming efficiency were observed (P > 0.05). CONCLUSION: Down-regulation of EphA2 gene by EphA2 siRNA in SKOV3 cell line showed retardation of cell growth, proliferation and partial reversion of the malignant phenotype, including the ability of adhesion and invasion. EphA2 may be a new and potent target of gene therapy in ovarian carcinoma.

Growth of hepatocellular carcinoma in the regenerating liver.

Liver resection and liver transplantation are the treatment modalities with the greatest potential for curing hepatocellular carcinoma (HCC). Tumor recurrence after resection for HCC is, however, a major problem, and an increased rate of recurrence after living donor transplantation versus cadaveric whole liver transplantation has been suggested. Factors involved in liver regeneration may stimulate the growth of occult tumors. The aim of this project was to test the hypothesis that a microscopic HCC tumor in the setting of partial hepatectomy would show enhanced growth and signs of increased invasiveness corresponding to the size of the liver resection. Hepatectomy was performed to various degrees in groups of Buffalo rats with the concomitant implantation of a fixed number of hepatoma cells in the remnant liver; a control group underwent only resection. After 21 days, the sizes and numbers of the tumors and the expression of alpha-fetoprotein (AFP), cyclin D1, calpain small subunit 1 (CAPNS1), CD34 (a microvessel density marker), vascular endothelial growth factor (VEGF), and vascular endothelial growth factor receptor 2 (VEGFR2) were evaluated and compared between the groups. The tumor volume and number increased significantly with the size of the partial hepatectomy (P < 0.05). The largest resections were also associated with increased hepatoma cell infiltration in the lungs and significant up-regulation of cyclin D1, AFP, CAPNS1, CD34, VEGF, and VEGFR2. The results suggest that liver regeneration after partial hepatectomy facilitates the growth and malignant transformation of microscopic HCC, and this could be significant for liver resection and partial liver transplantation strategies for HCC.

[Expression and prognostic significance of EphA2 and EphrinA-1 in ovarian serous carcinomas].

OBJECTIVE: To investigate the expression and prognostic significance of EphA2 and EphrinA-1 in ovarian serous carcinomas. METHODS: Ninety five tumors from the patients with ovarian serous carcinomas and 2 ovarian cancer cell lines were recruited. The expressions of EphA2 and EphrinA-1 were examined by means of immunohistochemistry. The relationships among protein expression and clinicopathological features, survival of patients were analyzed. The mRNA and protein expressions of EphA2 and EphrinA-1 in ovarian cancer cell lines were measured with semiquantitative polymerase chain reaction and western blotting. RESULTS: The protein expressions of EphA2 and EphrinA-1 in tumor were 92.6% (88/95)and 97.9% (93/95) respectively, while were 40%(8/20) and 30% (6/20) in adjacent ovarian tissue (P = 0.000). EphA2 immunohistochemical staining could be observed in both tumour and vascular endothelial cells, and EphrinA-1 mainly localized in the tumor cells. The expression of EphA2 was significantly associated with the expression of EphrinA-1 (r = 0.98, P = 0.02). There was no significant correlation between the expressions of EphA2/EphrinA-1 and age, FIGO stage, residual tumour size and histological grade. High levels of both EphA2 and EphrinA-1 protein expression were significantly associated with a shorter overall survival in multivariate analysis (P < 0.05). High levels of EphA2 and EphrinA-1 mRNA and protein were detected in OVCAR3 and SKOV3 cell lines. CONCLUSION: There are over-expression of EphA2 and EphrinA-1 in ovarian serous carcinomas and ovarian cancer cell lines. Tumours with higher expression levels of both EphA2 and EphrinA-1 significantly associated with poorer clinical outcome.

Dipeptidyl peptidase IV expression in cancer and stromal cells of human esophageal squamous cell carcinomas, adenocarcinomas and squamous cell carcinoma cell lines.

Dipeptidyl peptidase IV (DPPIV) is a transmembrane serine protease which is involved in the process of tumor invasion and development of metastases in human cancers. The aim of this study was to investigate the expression of DPPIV in cancer and stromal cells of both esophageal adenocarcinoma and squamous cell carcinoma (SCC). Tissue material from 159 patients was analyzed using immunohistochemistry. Western blotting was performed on cell lines and fresh frozen tissue sections. Results were compared with clinicopathological features. Evaluation of the immunohistochemical findings revealed significant differences between DPPIV expression in carcinoma cells and stromal cells, depending on the histological tumor type. A significantly higher level of DPPIV was found in adenocarcinomas compared to SCCs while no DPPIV was detected in normal esophageal epithelium. Overexpression of DPPIV in patients with adenocarcinoma was additionally associated with distant metastases. Thus, differences of DPPIV level in esophageal carcinomas compared with normal epithelium showed that esophageal malignancies were associated with an increased amount of cell surface-bound DPPIV. Radiotherapy in patients had no impact on DPPIV expression in analyzed tissue samples. There was no correlation between DPPIV expression in cancer or stromal cells and survival of the patients.

Seprase, dipeptidyl peptidase IV and urokinase-type plasminogen activator expression in dysplasia and invasive squamous cell carcinoma of the esophagus. A study of 229 cases from Anyang Tumor Hospital, Henan Province, China.

OBJECTIVE: Seprase, dipeptidyl peptidase IV (DPPIV) and urokinase-type plasminogen activator (uPA) play a crucial role in the degradation of the extracellular matrix and in the progression of various human tumors. However, their pathophysiologic significance in esophageal carcinoma has not yet been fully elucidated. METHODS: The expression of seprase, DPPIV and uPA in esophageal dysplasia, squamous cell carcinoma (SCC) and normal epithelium was examined by immunohistochemistry. RESULTS: Seprase, DPPIV and uPA immunoreactivity was found in dysplastic and cancer cells as well as in stromal cells adjacent to dysplasia and cancer sites, but not in normal epithelium. We found a significant association between uPA expression and sex, tumor size and histological classification in carcinomas. High expression of DPPIV in cancer cells correlated with longer survival of the patients. No significant associations between seprase and clinicopathological features either in dysplasia or in carcinomas were found. Finally, we demonstrated higher levels of seprase, DPPIV and uPA in SCC cell lines than in normal esophageal epithelial cell lines. CONCLUSIONS: Our results showed that seprase, DPPIV and uPA are expressed in both premalignant and malignant forms of SCC, but are lacking in normal esophageal epithelia, suggesting that they are involved in SCC neoplastic progression.

[Correlation of DPPIV expression with clinicopathological features and prognosis in epithelial ovarian carcinoma].

OBJECTIVE: To investigate the expression of dipeptidyl peptidase IV (DPPIV) in patients with epithelial ovarian carcinoma (EOC) and its clinical significance. METHODS: Immunohistochemistry (IHC) was used to detect the expression of DPPIV protein in 378 formalin-fixed paraffin-embedded EOC tissue samples. The expression of DPPIV mRNA in 86 EOC tissue samples were examined by in situ hybridization (ISH) using specific FITC-labelled RNA probes. Forty-two samples of normal ovarian tissues were used as control. Statistical analyses were carried out by Chi-square test, Spearman rank correlation and Kaplan-Meier method. RESULTS: Among the 378 epithelial ovarian carcinomas, 351 (92.9%) showed a positive expression of DPPIV protein, while only 25/42 (59.5%) of normal ovaries had a positive expression by semi-quantitative IHC analysis. The expression level of DPPIV protein was significantly lower in the normal ovaries than that in ovarian carcinomas (chi(2) = 18.4, P = 0.001). There was no significant correlation between the expression of DPPIV protein and age, FIGO stage and histological grade (P > 0.05). However, the expression of DPPIV protein was significantly associated with histological type (chi(2) = 28.5, P = 0.005). The patients with high level expression of DPPIV protein likely had a poor prognosis in terms of overall survival (P = 0.02). Of the 86 patients, 84 (97.7%)showed positive expression of DPPIV mRNA, also higher than that in normal ovarian tissues (P < 0.05). A statistically significant correlation between DPPIV mRNA and protein expression was observed (r(s) = 0.66, P = 0.001). CONCLUSION: DPPIV may be involved in the carcinogenesis of ovarian cancer, and may become a potential prognostic marker for epithelial ovarian carcinoma.

Expression of seprase in effusions from patients with epithelial ovarian carcinoma.

BACKGROUND: Seprase plays an important role in malignant cell invasion and metastasis by degrading the extracellular matrix. However, its clinical significance remains largely unknown. The objective of the current study was to evaluate the expression of seprase in effusions from patients with epithelial ovarian carcinoma and its clinical values. METHODS: Immunohistochemistry was used to examine the expression of seprase protein in a series of 74 malignant peritoneal (n = 64) and pleural (n = 10) effusions from Norwegian patients with epithelial ovarian carcinoma. Additionally, 34 effusions were evaluated using the Western blotting. Nine reactive effusions, obtained from patients with benign lesions, served as a control group. Statistical analyses were carried out by Chi-square test and Kaplan-Meier method. RESULTS: In the 74 malignant effusion specimens, 57 (77.02%) were positive for seprase, while only 2 (22.22%) of the control group were positively stained (P = 0.001). In the malignant effusions, 17 (22.97%), 22 (29.73%), 22 (29.73%), 13 (17.57%) had negative, weak, moderate and strong seprase protein expression, respectively. The expression of seprase protein was predominant in cytoplasm of carcinoma cells. Increased seprase protein was negatively associated with the overall survival rate of the patients (P = 0.03). However, there was no significant correlation between protein expression and FIGO stage, age, histology, and histological grade. By Western blotting, 27 of the 34 effusions showed the presence of both 170-kD dimeric form and 97-KD monomeric form of seprase while only 1 of the 34 had 170-KD dimeric form, which was consistent with the results of immunohistochemistry (P = 0.05). CONCLUSIONS: Seprase may be involved in the development of ovarian cancer, and is a potential predictive marker for the disease.

[Expressions of estrogen receptor subtypes in epithelial ovarian carcinomas].

OBJECTIVE: To investigate the expressions of estrogen receptor(ER) subtypes ERa and ERP in epithelial ovarian carcinomas. METHODS: One hundred and eighteen Norwegian patients with epithelial ovarian carcinoma were included in this study. The expressions of ERalpha and ERbeta were examined by means of immunohistochemistry. The relationships between protein expressions and clinicopathological features and survival were analyzed. Frozen tissues from 10 cases in which the tumors showed variable ERalpha and ERbeta protein expressions were used for Laser capture microdissection (LCM). Cancer cells in each frozen section were captured with the LCM method and processed for Western blot analysis. RESULTS: Of the 118 tumours, 32 (27.1%), 20 (16.9%), 17 (14.4%), 49 (41.5%) demonstrated negative, weak, moderate and strong expression of ERalpha protein, respectively; 1 (0.8%), 7 (5.9%), 13 (11%), 97 (82.2%) showed negative, weak, moderate and strong expressions of ERbeta protein, respectively. There was no significant association between ERalpha expression and the clinicopathological features such as age, histological type, FIGO stage, histological grade and residual tumour size. ERbeta expression was not associated with age, histological type,FIGO stage and residual tumour size, but it was significantly associated with higher histological grade. In addition, Kaplan-meier analysis revealed that high levels of ERbeta expression was significantly associated with a shorter overall survival (P = 0.03). CONCLUSION: There are ERalpha and ERbeta expressions in epithelial ovarian carcinomas. Higher level of ERbeta protein expression is associated with higher histological grade and poorer clinical outcome in the cases of ovarian carcinoma. ERbeta may be a useful marker of ovarian carcinogenesis and could predict the efficacy of endocrinotherapy and the prognosis for patients with ovarian carcinoma.

Dendritic and lymphocytic cell infiltration in prostate carcinoma.

We examined the distribution of CD1a⁺ cells and CD8⁺ and CD4⁺ T lymphocytes in prostate cancer (PCa) and correlated these with clinicopathological parameters. We also investigated whether the distribution of these cells was related to the expression of the cell membrane protein B7-H3, a putative negative regulator of the immune response expressed on PCa cells. A cohort of 151 PCa patients treated with radical prostatectomy (RP) was followed prospectively from 1985 until 2006 with a median follow-up of 9 years. Whole-mount sections of PCa specimens were immunostained to identify immune cells. A low number of CD1a⁺ cells was significantly associated with a high Gleason score and high pathological stage of pT3. The number of CD1a⁺ cells correlated significantly with the number of intratumoral and stromal CD8⁺ and stromal CD4⁺ lymphocytes. Kaplan-Meier analysis showed a tendency toward impaired biochemical progression-free survival in patients with few CD1a⁺ cells within their RP specimens. The expression of B7-H3 correlated inversely with the number of CD1a⁺ cells and intratumoral CD4⁺ lymphocytes; there was a trend for a similar inverse relationship between B7-H3 expression and the number of CD8⁺ lymphocytes.

[Correlation between methylation of 5'-CpG islands and inactivation of FHIT gene in cervical cancer].

BACKGROUND & OBJECTIVE: Fragile histidine triad (FHIT) gene, a tumor suppressor gene, correlates with tumorigenesis of many solid tumors, and may be inactivated via methylation. This study was designed to explore relationship of methylation of 5'-CpG islands with inactivation of FHIT gene in cervical cancer. METHODS: Methylation of 5'-CpG islands in 10 normal cervical squamous epithelial tissues, and 40 cervical cancer tissues was detected with methylation specific polymerase chain reaction (MSP), protein expression of FHIT was detected with immunohistochemistry, and their correlations with clinicopathologic features of cervical cancer were statistically analyzed. RESULTS: (1) The 5'-CpG islands methylation rate of FHIT gene in cervical cancer tissues was 40.0% (16/40), while no methylation of FHIT gene was found in normal cervical tissues. (2) The methylation rates of FHIT gene in cervical cancer of stage I was 14.3% (2/14), significantly lower than that in cervical cancer of stage II (56.5%, 13/23) (P<0.05). (3) Expression of FHIT protein in cervical cancer was 33.0% (12/40), significantly lower than that in normal cervical tissue (100%, 10/10) (P<0.05). CONCLUSION: The 5'-CpG islands methylation may play an important role in inactivation of FHIT gene, and may be related with tumorigenesis of cervical cancer.