Ge Lou

Discrimination between malignant and benign ovarian tumors by plasma metabolomic profiling using ultra performance liquid chromatography/mass spectrometry

BACKGROUND Discrimination between epithelial ovarian cancer (EOC) and benign ovarian tumor (BOT) has always been difficult in clinical practice. We investigated the application of metabolomics in distinguishing EOC and BOT and tried to discover valuable biomarkers. METHODS Plasma metabolomic profiling was performed using ultra-performance liquid chromatography mass spectrometry (UPLC/MS). Partial least-squares discriminant analysis was employed to classify EOC and BOT, and reveal their metabolic differences. The area under the receiver-operating characteristic curve (AUC) was utilized to evaluate the predictive performance of the metabolic profiles for external validation set. RESULTS The metabolomic profiles consisting of 535 metabolites revealed a clear separation between EOC and BOT, with AUC of 0.86 for the external validation set. 6 metabolic biomarkers were identified, and the plasma concentrations of the 4 ascertained biomarkers (L-tryptophan, LysoPC(18:3), LysoPC(14:0), and 2-Piperidinone) were lower in EOC patients than those in BOT patients. Among them, tryptophan and LysoPC have been suspected to participate in cancer progression, and 2-Piperidinone might be a novel biomarker for EOC. CONCLUSIONS Metabolomics could be used to discriminate EOC from BOT in clinical practice, and the identified metabolic biomarkers might be important on investigating the biological mechanisms of EOC.

miR-211 suppresses epithelial ovarian cancer proliferation and cell-cycle progression by targeting Cyclin D1 and CDK6

BackgroundEpithelial ovarian cancer (EOC) is a significant cause of morbidity and mortality. MicroRNAs play important roles in cancer development and progression. The microRNA miR-211 is localized on intron 6 of the Trpm1 gene at 15q13-q14, a locus that is frequently lost in neoplasms. Its function and loss-of-function have been described in normal and cancer cells and tissues. miR-211 is known to be dysregulated in ovarian cancer: however, its function and the downstream effect of its loss-of-function in ovarian cancer have not been described before.MethodsWe analyzed miR-211 expression in clinical samples of primary EOC tissues compared to normal epithelial ovarian tissues and in the EOC cell lines: OVCAR3, Caov3, OVCA429, SKOV3 and A2780 compared to human ovarian surface epithelial cells. We then investigated the effect of miR-211 on EOC cell proliferation and apoptosis by counting cell numbers, MTT, colony formation, cell cycle, and PI/Annexin V staining assays. A luciferase reporter system was developed to assess miR-211 regulation of the predicted targets. Expression level of discovered targets and correlation with miR-211 expression were analyzed in EOC tissues. Finally, OVCAR3 stably expressing miR-211 or control cells were injected subcutaneously into mice to determine in vivo effect of miR-211 on tumorigenesis.ResultsWe found that the expression of miR-211 is downregulated in EOC tissues and cell lines compared to normal epithelial ovarian tissue and human ovarian surface epithelial cells, respectively. miR-211 was found to arrest cells in the G0/G1-phase, inhibit proliferation and induce apoptosis. Cyclin D1 and CDK6 were found to be direct targets of miR-211, and when overexpressed in miR-211-expressing EOC cells, could restore proliferative ability. Finally, in vitro investigation confirmed that miR-211 is a tumor suppressor that controls Cyclin D1 and CDK6 expression.ConclusionsOur results demonstrate that miR-211 is a tumor suppressor that controls expression of Cyclin D1 and CDK6, and that its downregulation results in overexpression of Cyclin D1 and CDK6 which increases proliferation ability of EOC cells to proliferate compared to normal cells.

The long-term efficacy of neoadjuvant chemotherapy followed by radical hysterectomy compared with radical surgery alone or concurrent chemoradiotherapy on locally advanced-stage cervical cancer.

Objectives: The purpose of this study was to compare the long-term survival of patients with locally advanced cervical cancer (stages IB2-IIB) treated with neoadjuvant chemotherapy followed by radical hysterectomy (hysterectomy plus pelvic lymph node dissection) (NACT + RS) with the survival of those treated with radical surgery (hysterectomy plus pelvic lymph node dissection) (RS) or concurrent chemoradiotherapy (CCRT). Methods: A retrospective study was performed. Patients were followed up for 54 to 114 months (median, 82.8 months). All risk factors that may have affected the disease-free survival (DFS) and overall survival (OS) were assessed. Results: From January 2000 to December 2005, 476 eligible patients were followed up. The 5-year DFS rates of the NACT + RS, RS, and CCRT groups were 85.00%, 77.44%, and 52.94%, respectively (P < 0.0001), whereas the 5-year OS rates were 88.67%, 80.21% and 64.37%, respectively (P < 0.0001). The NACT + RS group had significantly higher survival rates than both the RS (DFS: hazard ratio = 1.870, P = 0.0031; OS: hazard ratio = 1.813, P = 0.0175) and CCRT (DFS: hazard ratio = 3.535, P < 0.0001; OS: hazard ratio = 3.157, P < 0.0001) groups, while adjusting for the pathological type, clinical stage, tumor size (initial), and age. The 5-year DFS rate for patients receiving TP (paclitaxel and cisplatin) was 90.55%, and 71.70% for patients receiving PVB (cisplatin, vincristine, and bleomycin); the 5-year OS rates were 96.75% for TP and 70.09% for PVB, respectively. Patients receiving TP had a statistically significant improvement in both 5-year DFS and OS rates (P < 0.001). Conclusions: Neoadjuvant NACT + RS improves the long-term DFS and OS of patients with locally advanced cervical cancer stage IB2-IIB compared with RS alone and especially compared with CCRT. In the NACT + RS group, NACT with TP improves the long-term DFS and OS of patients compared with patients who had PVB chemotherapy regimen. These results may provide some useful information for clinicians to treat patients with locally advanced cervical carcinoma.

Identification of metabolic biomarkers to diagnose epithelial ovarian cancer using a UPLC/QTOF/MS platform

Abstract Background. Currently available tests are insufficient to distinguish patients with epithelial ovarian cancer (EOC) from normal individuals. Metabolomics, a study of metabolic processes in biologic systems, has emerged as a key technology in the measurements of small molecular metabolites in tissues or biofluids. Material and methods. To investigate the application of metabolomics on selecting EOC-associated biomarkers, 173 plasma specimens (80 newly diagnosed EOC patients and 93 normal individuals) were analyzed using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/QTOF/MS). A two-step strategy was performed to select EOC-associated biomarkers. The first step was to select potential biomarkers in distinguishing 42 cancer patients from 58 normal controls through partial least-squares discriminant analysis (PLS-DA) and database searching, and the second step was to validate the discrimination performance of these biomarkers in a dataset contained 38 EOCs and 35 controls. Results. Eight candidate biomarkers were selected. The combination of these biomarkers resulted in the area of receiver operating characteristic curve (AUC) of 0.941, a sensitivity of 0.921, and a specificity of 0.886 at the best cut-off point for detecting EOC. Discussion. Our findings suggested that sharp differences in metabolic profiles exist between EOC patients and normal controls. The identified eight metabolites associated with EOC may be served as novel biomarkers for diagnosis.

Long noncoding RNA CCHE1 promotes cervical cancer cell proliferation via upregulating PCNA

Long noncoding RNAs (lncRNAs) have been shown to play important roles in carcinogenesis and progression. However, the roles and functional mechanisms of lncRNAs in cervical cancer remain largely unknown. In this study, we found that cervical carcinoma high-expressed lncRNA 1 (lncRNA-CCHE1) was significantly upregulated in cervical cancer tissues. The higher expression of CCHE1 was significantly correlated with large tumor size, advanced Federation of Gynecology and Obstetrics stage, uterine corpus invasion, and poor survival. Gain-of-function and loss-of-function experiments demonstrated that CCHE1 overexpression promotes the proliferation of cervical cancer cell. By contrast, the depletion of CCHE1 inhibits the proliferation of cervical cancer cells. RNA pull-down assays confirmed that CCHE1 physically associates with proliferating cell nuclear antigen (PCNA) messenger RNA, consequently enhances the expression of PCNA. The expression of CCHE1 and PCNA is significantly correlated in cervical cancer tissues. The depletion of PCNA abolishes the effects of CCHE1 on the proliferation of cervical cancer cells. Taken together, these findings indicate that CCHE1 plays a pivotal role in cervical cancer cell proliferation via increasing PCNA expression and serves as a potential prognostic biomarker and therapeutic target in human cervical cancer.

Large‐scale profiling of metabolic dysregulation in ovarian cancer

Ovarian cancer is the leading cause of death in gynecologic malignancies. Profiling of endogenous metabolites has potential to identify changes caused by cancer and provide inspiring insights into cancer metabolism. To systematically investigate ovarian cancer metabolism, we performed metabolic profiling of 448 plasma samples related to epithelial ovarian cancer (EOC) based on ultra‐performance liquid chromatography mass spectrometry in both positive and negative modes. These unbiased metabolomic profiles could well distinguish EOC from benign ovarian tumor (BOT) and uterine fibroid (UF). Fifty‐three metabolites were identified as specific biomarkers for EOC, and this is the first report of piperine, 3‐indolepropionic acid, 5‐hydroxyindoleacetaldehyde and hydroxyphenyllactate as metabolic biomarkers of EOC. The AUC values of these metabolites for discriminating EOC from BOT/UF and early‐stage EOC from BOT/UF were 0.9100/0.9428 and 0.8385/0.8624, respectively. Meanwhile, our metabolites were able to distinguish early‐stage EOC from late‐stage EOC with an AUC of 0.8801. Importantly, analysis of dysregulated metabolic pathways extends our current understanding of EOC metabolism. Metabolic pathways in EOC patients are mainly characterized by abnormal phospholipid metabolism, altered l‐tryptophan catabolism, aggressive fatty acid β‐oxidation and aberrant metabolism of piperidine derivatives. Together, these metabolic pathways provide a foundation to support cancer development and progression. In conclusion, our large‐scale plasma metabolomics study yielded fundamental insights into dysregulated metabolism in ovarian cancer, which could facilitate clinical diagnosis, therapy, prognosis and shed new lights on ovarian cancer pathogenesis.

MicroRNA-302b Suppresses Human Epithelial Ovarian Cancer Cell Growth by Targeting RUNX1

Background: The microRNA (miR)-302 family functions as a tumor suppressor in human cancer. However, its role in epithelial ovarian carcinoma (EOC) remains unknown. Here, we investigated the role of miR-302b and its target gene RUNX1 in EOC. Methods: The expression levels of miR-302b and RUNX1 were assessed by quantitative real-time PCR and western blotting. The effects of ectopic expression of miR-302b were evaluated by the MTT assay, colony forming assay and flow cytometry. RUNX1 was identified as a target of miR-302b and their interaction was confirmed by luciferase activity assays, RUNX1 silencing and overexpression of a RUNX1 mutant construct lacking the 3′UTR. The effect of miR-302b on the suppression of tumor growth was investigated in vivo in a xenograft mouse model. Results: MiR-302b levels were markedly decreased in EOC specimens. Ectopic expression of miR-302b in EOC cells inhibited cell proliferation and colony formation, induced G0/G1 arrest, and promoted apoptosis. RUNX1 was identified as a direct target of miR-302b, and knockdown of RUNX1 inhibited cell growth in a manner similar to miR-302b overexpression, whereas introduction of a 3′UTR mutant of RUNX1 reversed the suppressive effect of miR-302b. Furthermore, miR-302b overexpression led to the inactivation of the STAT3 signaling pathway in EOC cells and inhibited tumor growth in a xenograft mouse model. Conclusions: MiR-302b functions as a tumor suppressor in EOC by targeting RUNX1 and modulating the activity of the STAT3 signaling pathway.

Over‐expression of LAPTM4B is associated with poor prognosis and chemotherapy resistance in stages III and IV epithelial ovarian cancer

The purpose of this study was to determine whether LAPTM4B over‐expression is associated with the prognosis and chemotherapy resistance in patients with stages III and IV epithelial ovarian carcinoma, i.e., patients with peritoneal metastasis or lymph node metastasis of epithelial ovarian carcinoma.

LAPTM4B overexpression is a novel predictor of epithelial ovarian carcinoma metastasis

LAPTM4B is a novel tumor‐associated gene. To date, there have been no published data regarding the role of LAPTM4B expression in epithelial ovarian carcinoma metastasis. Therefore, this study was performed to determine whether LAPTM4B overexpression is a new predictor of epithelial ovarian carcinoma metastasis. LAPTM4B expression was evaluated in 22 normal ovarian specimens and 139 ovarian carcinomas by western blotting analyses and immunohistochemistry. Univariate and multivariate analyses were used to determine the association between LAPTM4B expression and epithelial ovarian carcinoma metastasis. Western blotting analysis demonstrated that LAPTM4B was overexpressed in metastatic tissues from patients with ovarian cancers, and immunohistochemistry results revealed that among 59 patients with LAPTM4B overexpression, 57 (96.6%) presented intraperitoneal metastasis and 31 (52.5%) had lymph node metastasis. The results of the univariate and multivariate analyses demonstrated that LAPTM4B overexpression correlated with metastasis. The odds ratio of high‐to‐low expression for intraperitoneal metastasis was 11.410 (95% CI: 2.357, 55.239) and that for lymph node metastasis was 6.332 (95% CI: 2.533, 15.831). For intraperitoneal metastasis, the sensitivity and specificity of LAPTM4B overexpression were 48.7% and 90.9%; for lymph node metastasis, they were 73.8%% and 71.1%, respectively. LAPTM4B overexpression is a new predictor of epithelial ovarian carcinoma metastasis and an important potential biomarker for the early diagnosis of ovarian carcinoma.

Correlation of TNFAIP8 overexpression with the proliferation, metastasis, and disease-free survival in endometrial cancer.

Tumor necrosis factor alpha-induced protein 8 (TNFAIP8) is an apoptosis regulator proven to have an important function in the proliferation, invasion, metastasis, and progression of malignancies. In this study, we investigated the clinical role of TNFAIP8 overexpression in endometrial cancer (EC) and determined the relationship of TNFAIP8 with the proliferative antigen Ki-67 and metastasis-related gene matrix metallopeptidase 9 (MMP9) in 225 tumor specimens by immunohistochemistry and western blot, in order to elucidate more information on the role of TNFAIP8 protein with regard to the pathogenesis of EC. An association was observed between TNFAIP8 overexpression and clinicopathologic factors, such as advanced International Federation of Gynecology and Obstetrics stage (P < 0.001), higher histologic grade (P = 0.017), deep myometrial invasion (P = 0.030), lymphovascular space invasion (P = 0.011), lymph node metastasis (P < 0.001), and recurrence. Furthermore, TNFAIP8 overexpression was strongly correlated with MMP9 and Ki-67 expression in the progression of ECs. Patients with high expression of TNFAIP8 (P < 0.001 for both) and Ki-67 (P = 0.007 and P = 0.008) had poor overall survival and disease-free survival (DFS) rates. MMP9 overexpression did not affect survival outcomes (P > 0.05). Multivariate Cox regression analysis revealed that TNFAIP8 (P = 0.029) and lymph node metastasis (P = 0.022) were independent factors of DFS in patients with EC. These findings suggested that TNFAIP8 may be used as a prognostic marker for the recurrence of EC, and its promotion of the proliferation and metastasis in EC may be due to its mediation of Ki-67 and MMP9.