Correlation analysis of cancer stem cell marker CD133 and human endogenous retrovirus (HERV)-K env in SKOV3 ovarian cancer cells.

BACKGROUND: Human endogenous retrovirus (HERV)-K is a type of retrovirus that is present in the human genome, and its expression is usually silenced in healthy tissues. The precise mechanism by which HERV-K env influences cancer stemness is not fully understood, but it has been suggested that HERV-K env may activate various signaling pathways that promote stemness traits in cancer cells. OBJECTIVE: To establish the connection between HERV-K env expression and cancer stemness in ovarian cancer cells, we carried out correlation analyses between HERV-K env and the cancer stem cell (CSC) marker known as the cluster of differentiation 133 (CD133) gene in SKOV3 ovarian cancer cells. METHOD: To perform correlation analysis between HERV-K env and CSCs, ovarian cancer cells were cultured in a medium designed for cancer stem cell induction. The expression of HERV-K env and CD133 genes was verified using quantitative real-time polymerase chain reaction (RT-qPCR) and Western blot analyses. Additionally, the expression of stemness-related markers, such as OCT-4 and Nanog, was also confirmed using RT-qPCR. RESULTS: In the stem cell induction medium, the number of tumorsphere-type SKOV3 cells increased, and the expression of CD133 and HERV-K env genes was up-regulated. Additionally, other stemness-related markers like OCT-4 and Nanog also exhibited increased expression when cultured in the cancer stem cell induction medium. However, when HERV-K env knockout (KO) SKOV3 cells were cultured in the same cancer stem cell induction medium, there was a significant decrease in the number of tumorsphere-type cells compared to mock SKOV3 cells subjected to the same conditions. Furthermore, the expression of CD133, Nanog, and OCT-4 did not show a significant increase in HERV-K env KO SKOV3 cells compared to mock SKOV3 cells cultured in the same cancer stem cell induction medium. CONCLUSION: These findings indicate that the expression of HERV-K env increased in SKOV3 cells when cultured in cancer stem cell induction media, and cancer stem cell induction was inhibited by KO of HERV-K env in SKOV3 cells. These results suggest a strong association between HERV-K env and stemness in SKOV3 ovarian cancer cells.

Effect of human endogenous retrovirus-K env gene knockout on proliferation of ovarian cancer cells.

BACKGROUND: Among various human endogenous retroviruses (HERVs), the HERV-K (HML-2) group has been reported to be highly related to cancer. In pancreatic cancer cells, shRNA-mediated downregulation of HERV-K env RNA decreases cell proliferation and tumor growth through the RAS-ERK-RSK pathway; in colorectal cancer, CRISPR-Cas9 knockout (KO) of the HERV-K env gene affects tumorigenic characteristics through the nupr-1 gene. OBJECTIVE: The effect of HERV-K env KO has not been studied in ovarian cancer cell lines. In this study, we analyzed the tumorigenic characteristics of ovarian cancer cell lines, including cell proliferation, migration, and invasion, and the expression patterns of related proteins after CRISPR-Cas9 KO of the HERV-K env gene. METHODS: The HERV-K env gene KO was achieved using the CRISPR-Cas9 system in ovarian cancer cell lines SKOV3 and OVCAR3. Tumorigenic characteristics including cell proliferation, migration, and invasion were analyzed, and related protein expression was investigated by western blot analysis. RESULTS: The expression of the HERV-K env gene in KO cells was significantly reduced at RNA and protein levels, and tumorigenic characteristics including cell proliferation, migration, and invasion were significantly reduced. In HERV-K env KO SKOV3 cells, the expression of the RB protein was significantly up-regulated and the cyclin B1 protein level was significantly reduced. In contrast, in HERV-K env KO OVCAR3 cells, the level of phospho-RB protein was significantly reduced, but other protein levels were not changed. CONCLUSION: The results of this study showed that HERV-K env gene KO affects cell proliferation, invasion, and migration of ovarian cells through RB and Cyclin B1 proteins, but the specific regulation pattern can differ by cell line.

Correlation of constitutive photomorphogenic 1 (COP1) and p27 tumor suppressor protein expression in ovarian cancer.

BACKGROUND: Constitutive photomorphogenic 1 (COP1) is an E3 ubiquitin ligase that regulates important target proteins for cell growth including p27. The tumor suppressor p27 negatively regulates the cell cycle by inhibiting cyclin-dependent kinase. COP1 negatively regulates p27 stability by mediating its nuclear export and degradation. OBJECTIVE: Even if COP1 and p27 are tightly related and have significant roles in tumor progression, the expression patterns and relationship of both proteins in cancer have not yet been studied. METHOD: We analyzed the expression patterns and relationship between COP1 and p27 using an ovarian cancer tissue microarray by dual immunofluorescence analysis. RESULTS: The expression levels of COP1 and p27 proteins were not significantly different between ovarian cancer tissue and normal control tissue. Other clinical data including age, tumor type, tumor grade, and stage were not significantly related to expression of the two proteins. The co-relationship between COP1 and p27 proteins was significantly high (Pearson correlation coefficient 0.79, p = 8.65 × 10-22). CONCLUSIONS: Our results demonstrate that while the expression levels of COP1 and p27 are highly correlated, they are not significantly related to cancer progression in ovarian cancer.

Diagnostic accuracy of inflammatory markers for distinguishing malignant and benign ovarian masses.

Objective: To evaluate the role of inflammatory markers for distinguishing malignant and benign ovarian masses. Methods: Preoperative demographic, clinicopathologic, and laboratory variables were reviewed in patients with an ovarian mass that was subsequently diagnosed as either epithelial ovarian cancer (EOC) or a benign ovarian mass on histologic analysis. The differences between variables of the two groups were further evaluated. Logistic regression analysis was applied to evaluate variables to predict the presence of EOC. Results: According to the analysis of 229 patients with EOC, 120 (52.4%) patients had serous adenocarcinoma. Of the 229 patients, 110 (48.1%) patients had stage I or II disease and 119 (52.0%) had stage III or IV disease. There was a significant difference between EOC and benign ovarian mass in median values of variables such as age, white blood cell (WBC) count, hemoglobin concentration, platelet count, cancer antigen 125 (CA125) levels, neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR) (all P < 0.001, except for WBC count [P = 0.009]). In addition, there was significant difference in median values of these continuous variables among early-stage EOC, advanced-stage EOC, and benign ovarian mass (P < 0.001 for all variables). On multivariate logistic regression analysis, age (odds ratio [OR] = 4.14, P < 0.001), CA125 levels (OR = 9.87, P < 0.001), NLR (OR = 1.76, P = 0.049), PLR (OR = 2.41, P = 0.004), and LMR (OR = 0.51, P = 0.024) were found to significantly predict the presence of EOC. Conclusion: The three LMR, NLR, and PLR markers were found to be predictors for the presence of EOC. Further prospective studies to assess these markers as screening tools for the presence of EOC are required.

Monocytosis as a prognostic factor for survival in stage IB and IIA cervical cancer.

Objective: To measure hematologic parameters derived from the white blood cell (WBC) count and differential count (DC) as prognostic factors for survival in patients with stage IB and IIA cervical cancer. Methods: We retrospectively examined demographic, clinicopathologic, and laboratory parameters in a cohort of 233 patients with International Federation of Gynecology and Obstetrics stage IB and IIA cervical cancer who underwent surgical resection. We further assessed the effects of the WBC count and DC-derived hematologic parameters on progression-free survival (PFS) and overall survival (OS) after controlling for other parameters. Results: Patients were followed up for a median of 46.6 months (range, 9-142 months). The Kaplan-Meier estimates of PFS and OS at 5 years were 88.5% and 92.3%, respectively. In a multivariate analysis, we identified the absolute monocyte count (AMC) (hazard ratio [HR], 11.78; P <0.001) and tumor size (HR, 5.41; P = 0.003) as the strongest prognostic factors affecting PFS. We also identified AMC (HR, 23.29; P <0.001), tumor size, (HR, 5.27; P = 0.033), and lymph node involvement (HR, 3.90; P = 0.027) as the strongest prognostic factors affecting OS. AMC remained prognostic with respect to PFS or OS in a Cox model that controlled for the neutrophil-lymphocyte ratio or lymphocyte-monocyte ratio, although neither ratio was a significant prognostic factor for survival. Conclusions: Monocytosis and an increased tumor size were found to be independent prognostic factors affecting both PFS and OS in patients with stage IB and IIA cervical cancer.

The Neutrophil-Lymphocyte Ratio Predicts Recurrence of Cervical Intraepithelial Neoplasia.

OBJECTIVE: The purpose of the present study was to determine the prognostic significance of the neutrophil-lymphocyte ratio (NLR) in recurrence of cervical intraepithelial neoplasia (CIN). METHODS: We evaluated the NLR as a prognostic marker in the entire cohort of 230 patients who had undergone surgical resection and were diagnosed with CIN. Subjects were categorized into two different groups based on the NLR (NLR-high and NLR-low) using cutoff values determined by receiver operating characteristic (ROC) analysis. The primary research objective for this study was to validate the impact of the NLR on recurrence-free survival (RFS) in patients with CIN. The secondary objective was to evaluate the impact of other hematologic parameters on RFS in CIN patients. RESULTS: Using the entire cohort, the most appropriate NLR cut-off value for CIN recurrence selected on the ROC curve was 2.1. The NLR-low and NLR-high groups included 167 (72.6%) and 63 patients (27.4%), respectively. According to Kaplan-Meier analysis, RFS rates during the entire follow-up period were considerably lower in the NLR-high group than in the NLR-low group (P = 0.0125). In multivariate survival analysis using Cox proportional hazard model, we identified the NLR, absolute eosinophil count (AEC), hemoglobin concentration, and mean corpuscular volume (MCV) as valuable prognostic factors that impact RFS. CONCLUSIONS: The NLR is an independent prognosticator for RFS following surgical resection in CIN patients. We also found that the AEC, hemoglobin level, and MCV were strongly associated with RFS, as determined by multivariate analysis using a Cox model. These hematological parameters might provide additional prognostic value beyond that offered by standard clinicopathologic parameters.

Clinical value of routine serum squamous cell carcinoma antigen in follow-up of patients with locally advanced cervical cancer treated with radiation or chemoradiation.

OBJECTIVE: The objective of this study was to evaluate the clinical benefits of routine squamous cell carcinoma antigen (SCC-Ag) monitoring of patients with locally advanced cervical squamous cell carcinoma treated with radiation or chemoradiation. METHODS: A total of 53 patients with recurrent cervical squamous cell carcinoma treated with radiotherapy or chemoradiation were enrolled in this study. A retrospective review of medical records was conducted. The role of routine monitoring of serum SCC-Ag was evaluated in terms of cost effectiveness and effect on survival after diagnosis of recurrence. RESULTS: Serum SCC-Ag abnormality (≥2.5 ng/mL) was observed in 62.3% of patients when recurrent disease was diagnosed. The first indicator of relapse was abnormal serum SCC-Ag level in 21 patients (39.6%), 10 of whom had asymptomatic recurrent disease amenable to salvage therapy. Adding SCC-Ag measurement to the basic follow up protocol improved the sensitivity for detecting recurrence (The sensitivity of the basic protocol vs. addition of SCC-Ag: 49.1% vs. 88.7%, P<0.001). Twenty-three patients who were candidates for salvage therapy with curative intent showed better survival compared with those who were not candidates for therapy (5-year survival: 36.6% vs. 0%, P=0.012). CONCLUSION: Surveillance with routine serum SCC-Ag monitoring can better detect asymptomatic recurrent disease that is potentially amenable to salvage therapy with curative intent. Early diagnosis of recurrent disease that can be treated with salvage therapy may lead to better survival.

The Lymphocyte-Monocyte Ratio Predicts Patient Survival and Aggressiveness of Endometrial Cancer.

OBJECTIVE: We assessed the prognostic implications of preoperative lymphocyte-monocyte ratio (LMR) in patients with endometrial cancer (EC). METHODS: We retrospectively examined the LMR as a prognostic variable in a cohort of 255 patients with EC who underwent surgical resection. Patients were categorized into two groups according to the LMR (LMR-low and LMR-high) using cutoff points determined by receiving operator characteristic (ROC) curve analysis. The primary objective was to correlate the LMR to clinicopathological factors; the secondary objective was to determine the survival significance of the LMR in patients with EC. RESULTS: Using data from the entire cohort, the most discriminative LMR cutoff value selected on the ROC curve was 3.28 for both disease-free survival (DFS) and overall survival (OS). The LMR-low and LMR-high groups included 33 (12.9%) and 222 patients (87.1%), respectively. The 5-year DFS rates in the LMR-low and LMR-high groups were 64.5 and 93.9% (P < 0.0001), respectively, and the 5-year OS rates in the two groups were 76.7 and 96.5% (P < 0.0001), respectively. On multivariate analysis, we identified histologic grade, International Federation of Gynecology and Obstetrics (FIGO) stage, and LMR levels as the strongest prognostic factors affecting DFS (P = 0.0037, P < 0.0001, and P < 0.0001, respectively), and FIGO stage and the LMR as the strongest prognostic factors predicting OS (P < 0.0001 and P < 0.0001, respectively). CONCLUSION: The LMR is an independent prognostic factor for both DFS and OS after surgical resection, and it provides additional prognostic value beyond standard clinicopathological parameters.

The Lymphocyte-Monocyte Ratio Predicts Patient Survival and Aggressiveness of Ovarian Cancer.

OBJECTIVE: To measure the prognostic value of the lymphocyte-monocyte ratio (LMR) in patients with epithelial ovarian cancer (EOC). METHODS: We retrospectively examined the LMR as a prognosticator in a cohort of 234 patients with EOC who underwent surgical resection. Patients were categorized into two different groups based on the LMR (LMR-low and LMR-high) using cut-off values determined by receiver operating characteristic (ROC) curve analysis. The objective of the study was to assess the effect of the LMR on progression-free survival (PFS) and overall survival (OS), and to validate the LMR as an independent predictor of survival. RESULTS: Using the data collected from the whole cohort, the optimized LMR cut-off value selected on the ROC curve was 2.07 for both PFS and OS. The LMR-low and LMR-high groups included 48 (20.5%) and 186 patients (79.5%), respectively. The 5-year PFS rates in the LMR-low and LMR-high groups were 40.0 and 62.5% (P < 0.0001), respectively, and the 5-year OS rates in these two groups were 42.2 and 67.2% (P < 0.0001), respectively. On multivariate analysis, we identified age, International Federation of Gynecology and Obstetrics (FIGO) stage, and cancer antigen 125 levels to be the strongest valuable prognostic factors affecting PFS (P = 0.0421, P = 0.0012, and P = 0.0313, respectively) and age, FIGO stage, and the LMR as the most valuable prognostic factors predicting OS (P = 0.0064, P = 0.0029, and P = 0.0293, respectively). Conclusion : The LMR is an independent prognostic factor affecting the survival of patients with EOC.

Intrapelvic dissemination of early low-grade endometrioid stromal sarcoma due to electronic morcellation.

Endometrioid stromal sarcoma is a rare malignancy that originates from mesenchymal cells. It is classified into low-grade endometrioid stromal sarcoma (LGESS) and high-grade endometrioid stromal sarcoma. Ultrasonographic findings of LGESS resemble those of submucosal myomas, leading to the possible preoperative misdiagnosis of LGESS as uterine leiomyoma. Electronic morcellation during laparoscopic surgery in women with LGESS can result in iatrogenic intraabdominal dissemination and a poorer prognosis. Here, we report a patient with LGESS who underwent a supracervical hysterectomy and electronic morcellation for a presumed myoma in another hospital. Disseminated metastatic lesions of LGESS in the posterior cul-de-sac and rectal serosal surface were absent on primary surgery, but found during reexploration. In conclusion, when LGESS is found incidentally following previous morcellation during laparoscopic surgery for presumed benign uterine disease, we highly recommend surgical reexploration, even when there is no evidence of a metastatic lesion in imaging studies.

Expression patterns of Thymosin ß4 and cancer stem cell marker CD133 in ovarian cancers.

Thymosin ß4 (Tß4), a small acidic actin binding peptide, is overexpressed in a side population of cancer stem cells and CD133-positive colorectal cancer stem cells. In order to understand the relationship between Tß4 and CD133, we studied the expression patterns of Tß4 and CD133 in ovarian cancers. The expression patterns of Tß4 and CD133 were studied in normal ovaries, primary ovarian cancers, metastatic ovarian cancers, primary stomach cancers, and normal stomachs by Western blot and immunohistochemistry. Expression patterns and co-localization of Tß4 and CD133 were examined by immunofluorescence and confocal laser-scanning microscopy. Tß4 is overexpressed in primary ovarian cancers, but not in primary stomach cancers, when compared with normal controls. However, Tß4 levels in metastatic stomach cancers to the ovary are significantly upregulated compared with levels in normal stomachs and primary stomach cancers. These results suggest that Tß4 levels are related to tumorigenesis in ovarian cancers and metastasis in stomach cancers. The expression of Tß4 in normal ovaries and normal stomachs was weak, but was co-localized with CD133 expression. Tß4 expression was also co-localized with CD133 expression in primary ovarian carcinomas, metastatic ovarian cancers from stomach cancers and primary stomach cancers. These data suggest that Tß4 expression is strongly related to CD133 expression and is a characteristic of stem cells or cancer stem cells.

A case of non-gestational choriocarcinoma arising in the ovary of a postmenopausal woman.

Primary ovarian choriocarcinoma arising from a germ cell is an extremely rare occurrence, especially in postmenopausal women, and the prognosis is poor. Non-gestational choriocarcinoma of the ovary (NGCO) accounts for 0.6% or less of all ovarian neoplasms. It is important to distinguish gestational choriocarcinomas of the ovary (GCO) from other carcinomas because of the poor prognosis of NGCO. We describe a case of NGCO with lung metastasis in a 55 year old woman, which we present together with a brief review of the literature.

A case of multiple metastatic low-grade endometrial stromal sarcoma arising from an ovarian endometriotic lesion.

The development of endometrial stromal sarcomas (ESSs) in foci of endometriosis is extremely rare, and few cases have been reported in the literature to date, particularly with regard to multiple extrauterine ESS. Here we report a case of endometrial stromal sarcoma with multiple metastasis that arose from an ovarian endometriotic lesion. The literature is also briefly reviewed.

Comparison of concurrent chemoradiotherapy with cisplatin plus 5-fluorouracil versus cisplatin plus paclitaxel in patients with locally advanced cervical carcinoma.

OBJECTIVE: The aim of this study was to compare survival outcomes and toxicities between concurrent radiotherapy with cisplatin plus 5-fluorouracil and that with cisplatin plus paclitaxel in patients with locally advanced cervical carcinoma. METHODS: We retrospectively reviewed data from 93 locally advanced cervical carcinoma patients (stage IB to IVA) who had been treated by concurrent radiotherapy with cisplatin plus 5-fluorouracil (CF, n=45) vs. cisplatin plus paclitaxel (CP, n=48) as primary therapy. Toxicities and survival outcomes were compared. RESULTS: In the CP group, there were higher frequencies of severe (grade 3 or 4) leukopenia (79.2%, as compared to 11.1% in the CF group), severe neutropenia (77.1%, as compared to 8.9% in the CF group) and severe peripheral neuropathy (12.5%, as compared to 2.2% in the CF group). In the CF group, there were higher frequencies of severe nausea (33.3%, as compared to 14.6% in the CP group) and severe hyponatremia (11.1%, as compared to 0% in the CP group). Five-year DFS of the CF and CP groups was 67.4% and 79.1%, respectively (p=NS). Five year OS of the CF and CP groups was 79.6% and 80.9%, respectively (p=NS). CONCLUSION: Concurrent radiotherapy with cisplatin plus paclitaxel showed increased leukopenia, neutropenia and peripheral neuropathy, but less gastrointestinal toxicity (nausea) than that with cisplatin plus 5-fluorouracil. Survival outcome between these two groups was not statistically different in this study. Large prospective randomized controlled studies will be needed to confirm this result.

The Correlation between the Scores of Written Examination, the Clinical Clerkship Examination, the Clinical Skill Assessment, and the Graduation Examination of the Medical Students.

PURPOSE: The aim of this study was to investigate correlations between medical student scores on 4 examinations: the written examination, clinical clerkship examination, clinical skill assessment, and graduation examination. METHODS: Scores for 51 students who entered Daegu Catholic Medical School in 2005 on the written examination, clinical clerkship examination, clinical skill assessment, and graduation examination were included. Correlations between the scores were analyzed statistically. RESULTS: The scores on the written examination showed a strong correlation with those of the clinical clerkship assessment (0.833) and graduation examination (0.821). The clinical clerkship assessment scores correlated significantly with graduation examination scores (0.907). In addition, clinical skill assessment scores correlated with the written examination (0.579), clinical clerkship examination (0.570), and graduation examination (0.465) scores. CONCLUSION: Overall, the correlation between the scores on the clinical clerkship examination and the written examination was more significant than the correlation between scores on the clinical clerkship examination and clinical skill assessment. Therefore, we need to improve the evaluation method for the clinical clerkship examination and clinical skill assessment.

Impact of Neoadjuvant Chemotherapy on Postoperative Morbidity in Locally Advanced Cervical Carcinoma.

PURPOSE: The purpose of this study was to test the hypothesis that neoadjuvant chemotherapy (NACT) does not increase morbidity in patients undergoing radical hysterectomy with lymphadenectomy for locally advanced cervical cancer. MATERIALS AND METGODS: A retrospective study was undertaken of 140 patients with locally advanced cervical cancer (FIGO stage Ia to IIb) who underwent radical hysterectomy with lymphadenectomy by the same surgeon at the same hospital. Among the 140 patients, 39 received NACT followed by radical hysterectomy with pelvic lymphadenectomy (NACT group). This group received three cycles consisting of cisplatin 100 mg/m2/day on day 1 and 5-fluorouracil 1000 mg/m2/day from day 1 to 5. The NACT group was compared, in terms of intraoperative morbidity and postoperative morbidity, with the other 101 patients who underwent radical hysterectomy with lymphadenectomy but without chemotherapy (surgery-only group). RESULTS: There were no significant differences in mean age, body weight or height between the two groups. The only significant difference was that the NACT patients had higher stages of cancer. The incidence of intraoperative morbidity did not differ between the NACT and surgery only patients. We considered the operation duration, amount of blood loss and need for transfusion as indicators of intraoperative morbidity. We could not find any significant differences in the duration of suprapubic catheterization, days of hemovac drainage, amount of drained hemovac fluid, days of hospitalization or postoperative febrile morbidity between the NACT and surgery-only groups. Patients in the surgery-only group had more postoperative complications (ureteral obstruction, intestinal obstruction, lymphocyst, lymphedema, and death) than the NACT group, although not to a statistically significant degree (P>0.05). CONCLUSION: In this retrospective review, there was no evidence that NACT increased intraoperative or postoperative morbidity in patients with locally advanced cervical cancer. As this was a retrospective study, other prospective, randomized studies are needed to confirm these results.