RCC2, a regulator of the RalA signaling pathway, is identified as a novel therapeutic target in cisplatin-resistant ovarian cancer.

Currently, cisplatin (DDP) is the first-line chemotherapeutic agent used for treatment of ovarian cancer, but gradually acquired drug resistance minimizes its therapeutic outcomes. We aimed to identify crucial genes associated with DDP resistance in ovarian cancer and uncover potential mechanisms. Two sets of gene expression data were downloaded from Gene Expression Omnibus, and bioinformatics analysis was conducted. In our study, the differentially expressed genes between DDP-sensitive and DDP-resistant ovarian cancer were screened in GSE15709 and GSE51373 database, and chromosome condensation 2 regulator (RCC2) and nucleoporin 160 were identified as 2 genes that significantly up-regulated in DDP-resistant ovarian cancer cell lines compared with DDP-sensitive cell lines. Moreover, RCC2, Ral small GTPase (RalA), and Ral binding protein-1 (RalBP1) expression was found to be significantly higher in DDP-resistant ovarian cancer tissues than in DDP-sensitive tissues. RCC2 plays a positive role in cell proliferation, apoptosis, and migration in DDP-resistant ovarian cancer cell lines in vitro and in vivo. Furthermore, RCC2 could interact with RalA, thus promoting its downstream effector RalBP1. RalA knockdown could reverse the effects of RCC2 overexpression on DDP-resistant ovarian cancer cell proliferation, apoptosis, and migration. Similarly, RalA overexpression could alleviate the effects of RCC2 knockdown in DDP-resistant ovarian cancer cells. Taken together, RCC2 may function as an oncogene, regulating the RalA signaling pathway, and intervention of RCC2 expression might be a promising therapeutic strategy for DDP-resistant ovarian cancer.-Gong, S., Chen, Y., Meng, F., Zhang, Y., Wu, H., Li, C., Zhang, G. RCC2, a regulator of the RalA signaling pathway, is identified as a novel therapeutic target in cisplatin-resistant ovarian cancer.

Ultrasound Features Improve Diagnostic Performance of Ovarian Cancer Predictors in Distinguishing Benign and Malignant Ovarian Tumors.

To determine whether ultrasound features can improve the diagnostic performance of tumor markers in distinguishing ovarian tumors, we enrolled 719 patients diagnosed as having ovarian tumors at Nanfang Hospital from September 2014 to November 2016. Age, menopausal status, histopathology, the International Federation of Gynecology and Obstetrics (FIGO) stages, tumor biomarker levels, and detailed ultrasound reports of patients were collected. The area under the curve (AUC), sensitivity, and specificity of the bellow-mentioned predictors were analyzed using the receiver operating characteristic curve. Of the 719 patients, 531 had benign lesions, 119 had epithelial ovarian cancers (EOC), 44 had borderline ovarian tumors (BOT), and 25 had non-EOC. AUCs and the sensitivity of cancer antigen 125 (CA125), human epididymis-specific protein 4 (HE4), Risk of Ovarian Malignancy Algorithm (ROMA), Risk of Malignancy Index (RMI1), HE4 model, and Rajavithi-Ovarian Cancer Predictive Score (R-OPS) in the overall population were 0.792, 0.854, 0.856, 0.872, 0.893, 0.852, and 70.2%, 56.9%, 69.1%, 60.6%, 77.1%, 71.3%, respectively. For distinguishing EOC from benign tumors, the AUCs and sensitivity of the above mentioned predictors were 0.888, 0.946, 0.947, 0.949, 0.967, 0.966, and 84.0%, 79.8%, 87.4%, 84.9%, 90.8%, 89.1%, respectively. Their specificity in predicting benign diseases was 72.9%, 94.4%, 87.6%, 95.9%, 86.3%, 90.8%, respectively. Therefore, we consider biomarkers in combination with ultrasound features may improve the diagnostic performance in distinguishing malignant from benign ovarian tumors.

Roflumilast enhances cisplatin-sensitivity and reverses cisplatin-resistance of ovarian cancer cells via cAMP/PKA/CREB-FtMt signalling axis.

OBJECTIVE: We previously demonstrated the roflumilast inhibited cell proliferation and increased cell apoptosis in ovarian cancer. In this study, we aimed to investigate the roles of roflumilast in development of cisplatin (DDP)-sensitive and -resistant ovarian cancer. METHODS: OVCAR3 and SKOV3 were selected and the corresponding DDP-resistant cells were constructed. Cell viability, proliferation, apoptosis, cycle were performed. Expression cAMP, PKA, CREB, phosphorylation of CREB and FtMt were detected. The roles of roflumilast in development of DDP-sensitive and -resistant ovarian cancer were confirmed by xenograft model. RESULTS: Roflumilast + DDP inhibited cell proliferation, and induced cell apoptosis and G0/G1 arrest in OVCAR3 and SKOV3 cells, roflumilast induced expression of FtMt, the activity of cAMP and PKA and phosphorylation of CREB in ovarian cancer cells and the above-effect were inhibited by H89. Downregulation of CREB inhibited the roflumilast-increased DDP sensitivity of ovarian cancer cells, and the roflumilast-induced FtMt expression and phosphorylation of CREB. Also, roflumilast reversed cisplatin-resistance, and induced expression of FtMt and activation of cAMP/PKA/CREB in DDP-resistant ovarian cancer cells. Similarly, treated with H89 or downregulation of CREB inhibited the changes induced by roflumilast. In vivo, roflumilast inhibited the development of SKOV3 or SKOV3-DDP-R xenograft models. CONCLUSIONS: Roflumilast enhanced DDP sensitivity and reversed the DDP resistance of ovarian cancer cells via activation of cAMP/PKA/CREB pathway and upregulation of the downstream FtMt expression, which has great promise in clinical treatment.

Combined overexpression of cadherin 6, cadherin 11 and cluster of differentiation 44 is associated with lymph node metastasis and poor prognosis in oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is a highly invasive lesion that frequently metastasizes to the cervical lymph nodes and is associated with a poor prognosis. Several adhesion factors, including cadherin 6 (CDH6), cadherin 11 (CDH11) and cluster of differentiation 44 (CD44), have been reported to be involved in the invasion and metastasis of multiple types of cancer. Therefore, the aim of the present study was to determine the expression of CDH6, CDH11 and CD44 in tumor tissues from patients with OSCC, and whether this was associated with the metastasis and survival of OSCC. The mRNA expression of the human tumor metastasis-related cytokines was examined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) in OSCC tumors with or without lymph node metastasis (n=10/group). The expression of CDH6, CDH11 and CD44 in 101 OSCC and 10 normal oral mucosa samples was examined by immunohistochemical staining. The association between overall and disease-specific survival times of patients with OSCC and the expression of these three proteins was evaluated using Kaplan-Meier curves and the log-rank test. RT-qPCR results indicated that the mRNA expression of CDH6, CDH11 and CD44 was increased in OSCC patients with lymph node metastasis (2.93-, 2.01- and 1.92-fold; P<0.05). Overexpression of CDH6, CDH11 and CD44 was observed in 31/35 (89%), 25/35 (71%) and 31/35 (89%) patients, respectively. The number of OSCC patients with lymph node metastasis exhibiting CDH6, CDH11 and CD44 overexpression was significantly higher than the number of patients without lymph node metastasis exhibiting overexpression of these proteins (P=0.017, P=0.038 and P=0.007, respectively). OSCC patients with high co-expression of CDH6, CDH11 and CD44 exhibited lower disease-specific survival times (P=0.047; χ2=3.933) when compared with OSCC patients with low co-expression of these adhesion factors. CDH6, CDH11 and CD44 serve important roles in OSCC metastasis and the combined use of these factors as biomarkers may improve the accuracy of the prediction of cancer metastases and prognosis.

Roflumilast restores cAMP/PKA/CREB signaling axis for FtMt-mediated tumor inhibition of ovarian cancer.

The abrogation of cAMP generation by overexpression of PDE isoforms promotes the inflammatory pathology, and the PDE inhibitors have showed the potential anti-inflammation effects in clinical. However, the function of PDE inhibitors in cancer treatment remains unclear. We here investigated the role of PDE4 inhibitor Roflumilast in the treatment of ovarian cancer. We found that Roflumilast could effectively inhibit the proliferation, and induce apoptosis and cell cycle arrest in two ovarian cancer cell lines OVCAR3 and SKOV3. Meanwhile, the cAMP/PKA/CREB signals was activated by Roflumilast, which was accompanied by the up-regulation of mitochondrial ferritin (FtMt) level. Interestingly, forced expression of FtMt in ovarian cancer enhanced the apoptosis and inhibited tumor growth and the PKA inhibitor H89 and knockdown of CREB significantly repressed the expression of FtMt to restore the tumor proliferation and inhibit apoptosis. In addition, we found that Roflumilast-induced phosphorylated CREB directly promoted transcription of FtMt, indicating that Roflumilast up-regulated the expression of FtMt in ovarian cancer via cAMP/PKA/CREB signals. The anti-tumor role of Roflumilast in vivo was also demonstrated, the treatment of roflumilast effectively inhibited tumor proliferation and elevated the FtMt expression to restrict the tumor growth via the activation of cAMP/PKA/CREB signals in ovarian cancer.

[Diagnostic value of CA125, HE4 and Copenhagen Index in differentiating benign from malignant epithelial ovarian tumors].

OBJECTIVE: To analyze diagnostic value of Copenhagen Index based on pretreatment serum CA125, HE4 and age in differentiating benign and malignant epithelial ovarian tumors. METHODS: The clinical data were analyzed for 208 consecutive patients with epithelial ovarian tumors (including 100 with malignant and 108 with benign tumors) treated in our department between September, 2014 and September, 2016. The receiver-operating characteristic curve was drawn based on the golden standard of pathological diagnosis for calculation of the diagnostic sensitivity and specificity of CA125, HE4 and the Copenhagen Index. RESULTS: In the overall cases, early stage cases and advanced stage cases, the prediction probabilities of CA125, HE4 and Copenhagen Index were all significantly higher for malignant than in benign tumors (P<0.001). The sensitivities of CA125, HE4, Copenhagen Index for differentiating benign and malignant tumors were 81.0%, 86.0% and 91.0% in the overall cases, 64.0%, 68.0% and 72.0% in early stage cases, and 86.7%, 92.0% and 97.3% in advanced stage cases, and their diagnostic specificities were 88.0%, 93.5% and 96.3%, respectively. Copenhagen Index had the highest diagnostic sensitivity (but not in early stage cases) and specificity followed by HE4 and then by CA125 (P<0.001) (P>0.05). CONCLUSION: Copenhagen Index combined with CA125, HE4 and age hase better diagnostic value than HE4 or CA125 alone for differentiation between benign and malignant epithelial ovarian tumors, and can be used clinically to improve the early diagnostic rate of epithelial ovarian cancer.

[Effects of docosahexaenoic acid on cell apoptosis, invasion and migration of cervical cancer cells in vitro].

OBJECTIVE: To investigate the effect of docosahexaenoic acid (DHA) on apoptosis, migration and invasion of cervical cancer cell lines. METHODS: cervical cancer cell lines Hela and Siha in logarithmic phase were treated different concentrations of DHA. The morphological changes of the cells were observed microscopically and cell apoptosis was observed using Hoechst 33258 fluorescent staining. MTT assay was used to evaluate the effect of DHA in suppressing cell growth, and flow cytometry was employed to analyze the changes of cell apoptotic rate following DHA stimulations. Wound healing assay and Transwell migration assay were used to evaluate the migration of the cell lines. The expression levels of Bax, Bcl-2 cleaved caspase3, MMP-9 and VEGF proteins were detected by Western blotting. RESULTS: DHA exposure of the cells caused obvious morphological changes and dose-dependently increased the number of apoptotic bodies in the cells. MTT assay showed that DHA inhibited the growth of the cancer cells in a time- and concentration-dependent manner. DHA also effectively suppressed migration and invasion of the cancer cells. The cells exposed to DHA showed significantly down-regulation of Bcl-2, MMP-9 and VEGF proteins and up-regulation of cleaved-caspase 3 and Bax. CONCLUSION: DHA can promote cervical carcinoma cell apoptosis by down-regulating the anti-apoptotic proteins Bax, Bcl-2 and cleaved-caspase3 and suppress cell invasion by decreasing MMP-9 and VEGF expressions.

[One-stage reconstruction of oral and maxillofacial soft tissue defect by using forearm free flap].

OBJECTIVE: To study technique of one-stage reconstruction of oral and maxillofacial soft tissue defect by using forearm free flap. METHODS: The microsurgery procedure was used in the treatment of 4 oral cancer patients with oral and maxillofacial soft tissue defect after surgery. RESULTS: The forearm free flap was survived, for 4-18 months the oral function was well, and the appearance was satisfactory. CONCLUSION: This is a good approach to the reconstruction of oral and maxillofacial soft tissue defect by using forearm free flap technique.

[The application of free tissue flap in simultaneous reconstruction of defects after tongue cancer].

OBJECTIVE: To discuss the feasibility of using free tissue flap for simultaneous tongue reconstruction after tongue cancer resection. METHOD: Radial forearm flaps and anterolateral thigh flaps were used for simultaneous reconstruction of different defects after tongue cancer resection in 47 cases. The functional evaluation was assessed during the follow-up period of 3 months to 9 years. RESULTS: Only one flap failed,giving a success rate of 97.7%. The 36 patient available for postoperative follow-up were all able to communicate in basic languages and eat normally. Among them, normal speech was found in 34, while slurred speech was found in two. 31 patients could eat normal diet ,while eight could eat soft diet and two could eat liquid diet. CONCLUSIONS: Simultaneous tongue reconstruction with free tissue flap is a reliable method with high successful rate. Flaps selection based on different tongue defects is the key point to achieve good functional and cosmetic results for the reconstruction tongue.

[Mutation and expression of tumor suppressor gene phosphatase and tensin homolog deleted in chromosome 10 in oral squamous cell carcinoma].

OBJECTIVE: To investigate the role of mutation and mRNA expression of tumor suppressor gene phosphatase and tensin homolog deleted in chromosome 10 (PTEN) in tumorigenesis and progression of oral squamous cell carcinoma (OSCC). METHODS: The mutation of exon 3, 5, 6 and exon 8 of PTEN gene in 42 oral squamous cell carcinoma tissue and paired normal tissue were detected by polymerase chain reaction (PCR) and DNA sequencing methods. The levels of PTEN mRNA expression in these tissues were assayed using semi-quantitative RT-PCR. Data was analyzed with SPSS 14.0 software package. RESULTS: Mutated exon 5 of PTEN gene was found in 2 cases of advanced OSCC. The expression of PTEN mRNA was detected in all OSCC and paired normal tissue. The level of PTEN mRNA in OSCC tissue (0.36 +/- 0.12) was significantly lower than that of paired normal tissue (0.64 +/- 0.09, P < 0.01). CONCLUSIONS: The decreased expression of PTEN mRNA contributes to tumorigenesis of OSCC.

[Reconstruction of large through-and-through palate defects with folded free forearm flap].

OBJECTIVE: To explore the reconstruction method of large through-and-through palate defects. METHODS: From 2003 to 2006, 7 cases of large through-and-through palate defects were reconstructed with vascularized folded free forearm flap. 8 flaps were used, including 7 free forearm flaps and 1 pectoralis major myocutaneous flap. RESULTS: All the tissue flaps survived well except one flap necrosis because of arterial thrombosis. The appearance of reconstructed palate was acceptable, and the functions of swallowing and speech were normal or almost normal. CONCLUSIONS: It is feasible and effective to repair large through-and-through palate defects with folded free forearm flap.

[Branchial cleft anomalies: a review of 40 cases].

OBJECTIVE: To explore the diagnosis and treatment for the congenital branchial cleft cysts and sinuses. METHOD: We analyzed the clinical material of 40 patients with branchial cleft cysts and sinuses, and explored the diagnosis and therapeutic experience. Twelve cases were first branchial cleft cysts and sinuses, 23 cases were second branchial cleft cysts and sinuses and 5 cases were third branchial cleft cysts and sinuses. RESULT: All patients were cured by surgical resection. Followed up from 1 to 5 years, 36 cases were cured, 2 cases were lost touch with, 2 cases recurred and were well treated again. CONCLUSION: Surgical resection was the main method to treat the congenital branchial cleft cysts.

[Application of CTA and CAD\CAM techniques in mandible reconstruction with free fibula flap].

OBJECTIVE: To explore the application of computed tomographic angiography (CTA) and computer aided design\manufacture (CAD\CAM) techniques in mandible reconstruction with free fibula flap. METHODS: 7 cases of complex mandible defect were reconstructed with free fibula flap. Patients' lower extremity vasculature was evaluated by preoperative CTA examination; mandible tumor resection and reconstruction were simulated by CAD technique, the surgical template was manufactured with CAM technique preoperatively; and fibula flap was shaped according to surgical template during mandible reconstruction operation. RESULTS: Preoperative CTA examination performed without complication, the desired anatomy was adequately demonstrated in all patients. All of flaps survived, patients' facial contour are symmetry approximately. CONCLUSIONS: CTA and CAD\CAM techniques have important value for complex mandible reconstruction in selection of donor site, planning of operation and shaping of fibula flap.

[Diagnosis and treatment of Madelung's disease].

OBJECTIVE: To review the characteristics of Madelung's disease which is rare and unfamiliar to clinicians and to find the method of diagnosis and treatment. METHODS: Detailed clinical data of 7 patients with Madelung's disease were reviewed and analyzed. And related literatures were discussed together. RESULTS: All of 7 patients have excessive subcutaneous fat deposit predominantly around neck. One patients had the complication of central and peripheral neuropathy, One patients had the complication of glucose intolerance, One patients had the complication of chronic hepatopathy and hyperuricaemia. These 3 patients were associated with sleep apnea syndrome simultaneously. Another one patient was accompanied by autonomic nervous system disease only. Total neck lipectomy and abstinence from alcohol were performed on 5 patients. No recurrence was seen during a follow-up of 6 months to 5 years. One patient received partial lipectomy twice and relapsed again. All pathological results were nonencapsulated fat. One patient refused treatment. Six men patients were alcohol abusers. CONCLUSIONS: Madelung's disease is characterized by massive accumulation of nonencapsulated subcutaneous fat mainly located symmetrically in the neck. Chronic alcoholism may be a major risk factor. It may be associated with some internal diseases. Total neck lipectomy to improve figuration and relieve pressure and abstinence from alcohol may be the main effective therapy.