Prognostic factors of 87 ovarian yolk sac tumor (OYST) patients and molecular characteristics of persistent and recurrent OYST.

OBJECTIVE: We aimed to explore the characteristics of OYST, particularly for persistent and recurrent OYST, in order to explore potential treatment options and thereby improve patient outcomes. METHODS: We retrospectively reviewed the clinical records of all patients with OYST at Fudan university Shanghai Cancer Center from December 3, 2005 to November 27, 2020. Furthermore, and performed whole-exome sequencing on 17 paired OYST (including 8 paired persistent and recurrent OYST) tumor and blood samples to elucidate the aberrant molecular features. RESULTS: Totally, 87 OYST patients were included between 2007/03/13 and 2020/11/17. With a median follow-up of 73 [3-189] months, 22 patients relapsed or disease persisted. Overall, 17 patients died with a median overall survival of 21 [3-54] months. Univariate and multivariate analysis revealed tumor histology and residual lesions were independently associated with event free survival and overall survival, cycles to AFP normalization were another independent risk factor for overall survival. For the 8 persistent and recurrent OYST: cancer driver genes including ANKRD36, ANKRD62, DNAH8, MUC5B, NUP205, RYR2, STARD9, MUC16, TTN, ARID1A and PIK3CA were frequently mutated; cell cycle, ABC transporters, HR, NHEJ and AMPK signal pathway demonstrated as the most significantly enriched pathways; TMB, DNA MMR gene mutation and MSI were significantly higher. Mutation signature 11, 19 and 30 were the dominant contributors in persistent and recurrent OYST mutation. CONCLUSION: Persistent and recurrent OYST associated with poor prognosis, and probably susceptible to immune checkpoint blockade therapy. Molecular characteristics contributed to predict the persistence and recurrence of OYST.

Surgical and oncologic outcomes between laparoscopic and radical abdominal hysterectomy for IB1-IIA2 cervical cancer.

OBJECTIVE: To compare sugrical and survival outcomes between laparoscopic radical hysterectomy (LRH) and radical abdominal hysterectomy (RAH). METHODS: All the patients with IB1-IIA2 cervical cancer who performed LRH or RAH in Fudan University Shanghai Cancer Center between 1/2016 and 12/2017 were retrospectively analyzed. RESULTS: There were no significant differences between LRH and RAH groups except deep stromal invasion (35.2% vs 54.4%, p = 0.000), operating time (232.3 ± 61.9 min vs. 106.7 ± 36.2 min, p = 0.000), blood loss (169.5 ± 96.2 ml vs. 219.6 ± 149.3 ml, p = 0.000), and lymph node counts (21.1 ± 7.1 vs. 23.2 ± 8.7 min, p = 0.012). The LRH group displayed poorer disease-free survival (DFS) (5-year rate, 79.4% vs. 90.0%; p = 0.046) and overall survival (OS) (5-year rate, 74.7% vs. 90.0%; p = 0.026) compared to the RAH group. On multivariate analysis, LRH was an independent risk factor for DFS (hazard ratio, 0.377; 95% confidence interval [CI], 0.227-0.625; p = 0.000) and OS (hazard ratio, 0.434; 95% CI, 0.254-0.740; p = 0.003). CONCLUSIONS: LRH affected the survival of cervical cancer patients with tumor size >2 cm (p < 0.05). Adjuvant therapy could not improve the prognosis of laparoscopic patients (p < 0.05).

Hybrid Capture-based Genomic Profiling of Circulating Tumor DNA From Patients With Advanced Ovarian Cancer.

Objective: We conducted this study to characterize somatic genomic alterations in circulating tumor DNA (ctDNA) from patients with ovarian cancer and compare GAs detected in ctDNA with tissue databases. Methods: Hybrid capture-next generation sequencing genomic profiling of 150 genes was performed on ctDNA from 138 patients with ovarian cancer with 1,500× sequencing depth. The GAs detected in ctDNA were compared with those in our ovarian cancer tissue database (N = 488) and the Cancer Genome Atlas (TCGA) database (N = 489). Results: 115 patients (83%) had at least 1 GA detected in ctDNA. The most frequently altered genes detected in ctDNA were TP53 (72%), KRAS (11%), LRP1B (10%), ZNF703 (9%) and NF1 (8%). Comparative analysis with our tissue database showed similar frequencies of GAs per gene, although PIK3CA and KRAS mutations were more frequent in tissue and ctDNA, respectively (p < 0.05). Gene amplification and rearrangement were more frequent in ctDNA samples. The mutation frequency of homologous recombination repair associated-genes, VEGF signal/angiogenesis pathways, RAS pathways, NOTCH pathways and MSI-H ratio was not statistically different either in ctDNA or in tissue database. However, the mutation frequency of AKT, PIK3CA, PTEN and STK11 in PI3K/AKT/mTOR pathway was significantly lower than that in tissue samples (p < 0.05). Conclusions: Our results suggest that genomic profiling of ctDNA could detect somatic GAs in a significant subset of patients with ovarian cancer. Hybrid capture-NGS based on liquid biopsy has the potential capability to serve as a substitute to tissue biopsy and further studies are warranted.

Promoter Methylation-mediated Silencing of the MiR-192-5p Promotes Endometrial Cancer Progression by Targeting ALX1.

Background: Epigenetic regulation by promoter methylation-mediated silencing of cancer-related microRNAs plays vital roles in tumorigenesis. MiR-192-5p promotes tumor progression in various human cancers with conflicting biological effects. However, its expression levels and biological functions in endometrial carcinoma (EC) have not been reported. Methods: The methylation status of miR-192-5p in tissue samples and cell lines, was examined using bisulfite sequencing PCR. miR-192-5p expression was also measured. EC cell lines transfected with specifically designed vectors overexpressing miR-192-5p, its target gene ALX1 or both, were constructed. Tumorigenicity of these cell lines were examined by in vitro and in vivo experiments. Dual-luciferase reporter assay were employed to verify the target of miR-192-5p. Results: The promoter region of miR-192-5p gene was highly methylated and its expression significantly repressed in EC samples. Moreover, a higher level of promoter methylation as well as a lower expression of miR-192-5p, was significantly associated with advanced Federation of Gynecology and Obstetrics stage and shorter disease-free survival in patients with curatively resected EC. Functional studies demonstrated that miR-192-5p overexpression inhibited in vitro tumor progression, in vivo tumorigenicity and the expression of several oncoproteins that was highly related to epithelial-to-mesenchymal transition. ALX1 was verified as a direct target of miR-192-5p and demonstrated to mediate the tumor-suppressive function of miR-192-5p. Conclusion: miR-192-5p is a tumor suppressor miRNA that is epigenetically silenced by promoter methylation and may serve as a potential prognostic biomarker in EC.

Clinical characteristics and risk factors of invasion in extramammary Paget's disease of the vulva.

PURPOSE: This study aimed to evaluate the risk factors of recurrence and invasive disease in patients with extramammary Paget's disease of the vulva (EPDV). METHODS: We performed a retrospective analysis of patients who were initially diagnosed with EPDV in Fudan University Shanghai Cancer Center between May 2006 and March 2019. RESULTS: Thirty-eight patients were initially diagnosed with EPDV in our institution. Among them, 29 had intraepithelial EPDV, 8 had intraepithelial EPDV with stromal invasion, and 1 had an underlying vulvar adenocarcinoma. In total, 8 (21%) patients had 12 recurrences. Of these eight patients, four had one recurrence, while other four had two recurrences. Intraepidermal EPDV recurred nine times, while intraepidermal EPDV with invasive disease recurred thrice. The first and second recurrence intervals were 62.1 (9-146) months and 22 (15-28) months, respectively. The rate of invasive disease was 23.7% (9/38) for primary EPDV and 25% (3/12) for recurrent ones. We determined that the presence of invasive disease was associated with a history of more than 10 years (p = 0.02) and inversely correlated with positive margins (p = 0.037), However, invasive disease had no statistical relations with age (p = 0.438), recurrence (p = 0.642), and lesion diameter (p = 0.08). CONCLUSIONS: EPDV with a history of more than 10 years was associated with invasive disease. Close and long-term follow-up are recommended to identify those who require further treatment.

A preoperative prediction model for predicting coexisting adnexa malignancy of patients with G1/G2 endometrioid endometrial cancer.

OBJECTIVE: To identify the predictors of coexisting adnexa malignancy (CAM) before surgery for patients with G1/G2 endometrioid endometrial cancer (EEC). METHODS: Patients with G1/G2 EEC who received surgery in Fudan University Shanghai Cancer Center from 1996 to 2017 were enrolled. Univariate and multivariate logistic regression were performed to identify the predictors for CAM, and the nomogram was constructed and evaluated the discrimination and calibration. RESULTS: Among the 1511 patients in the study cohort, 66 (4.4%) coexisted adnexa malignancy (51 metastatic and 15 synchronous primaries). In the univariate logistic regression analysis, CA125 level (>35 U/ml), histologic grades, myometrial invasion depth in magnetic resonance imaging (MRI), adnexal involvement in MRI/surgical exploration (SEP) were found to be significant predictors for CAM (P < .001, 0.047, 0.011, <0.001, respectively). The multivariate analysis demonstrated that high CA125 level (P < .001; OR: 2.945; 95%CI: 1.700-5.101), deep myometrial invasion (P = .011; OR: 2.194; 95%CI: 1.200-4.011), and suspected adnexal involvement in MRI/SEP (P < .001; OR: 11.524; 95%CI: 6.726-19.744) were independent predictors for CAM (AUC = 0.786). In 338 patients with MMR results, eighty-seven (25.7%) were detected MSI-high. There were 5.7% (5/87) patients diagnosed with CAM in the MSI-high group compared with 4.4% (11/251) in the MSS group. CONCLUSIONS: A nomogram with pre- and intra-operative factors was constructed to predict CAM in G1/G2 EEC patients, which may help clinicians in decision-making for ovarian preservation for these patients.

Incidence and risk factors of preoperative venous thromboembolism and pulmonary embolism in patients with ovarian cancer.

INTRODUCTION: To determine the incidence and predisposing factors of preoperative venous thromboembolism (VTE), especially pulmonary embolism (PE) in patients with ovarian cancer. MATERIAL AND METHODS: This retrospective study included 387 patients with primary ovarian cancer, whose preoperative work up included both ultrasonography of lower extremity vein and spiral computed tomography pulmonary angiogram, from September 2013 to November 2016. SPSS 22 was used for statistical analyses. RESULTS: The incidence of preoperative VTE and PE was 13.4% (52 patients), 9.3%(36 patients), respectively. Both the univariate and multivariate analyses revealed that D-dimer (DDI) level, age, and massive ascites were associated with preoperative VTE. Moreover, DDI level (odds ratio [OR] 3.133, 95% confidence interval [CI] 1.193-8.225, p = .02), massive ascites (OR 9.972, 95% CI 3.687-26.968, p < .001), and preoperative deep vein thrombosis (OR 5.977, 95% CI 2.073-17.228, p = .001) were independent predictors for preoperative PE. Additionally, the incidence of preoperative VTE and PE increased with increasing DDI levels (p < .001). DDI level < 1.5 µg/ml has 100% negative predictive value and 100% sensitivity for diagnosis of preoperative VTE and PE; however, DDI >5 µg/ml, the incidence of preoperative VTE and PE were 18.4% and 14.2% respectively. Moreover, DDI value was significantly correlated with preoperative PE volume (r = 0.746, p < .001). CONCLUSION: Preoperative VTE and PE are common events in patients with ovarian cancer. DDI level is a useful parameter for diagnosing and evaluating preoperative VTE and PE.

Methylation­associated silencing of miR­638 promotes endometrial carcinoma progression by targeting MEF2C.

Promoter methylation­associated silencing of cancer­associated microRNAs (miRNAs) is a common epigenetic mechanism during tumorigenesis in various types of human cancer. However, this has not been comprehensively examined in endometrial carcinoma (EC). In the present study, an miRNA microarray consisting of 1,347 common human miRNAs was used to select potential tumor suppressive miRNAs that were hyper­methylated in EC. This led to the identification of miR­638, miR­210 and miR­3665. The methylation status of miR­638 was examined by bisulfite sequencing polymerase chain reaction and miR­638 expression was measured by TaqMan miRNA assays. EC cell lines transfected with vectors overexpressing miR­638, its target gene myocyte enhancer factor 2C (MEF2C) or both, were constructed. Dual­luciferase reporter assays, a xenograft mouse model and rescue experiments were designed to study miR­638 and its target gene MEF2C. The results indicated that the promoter region of miR­638 was highly methylated and the expression of miR­638 was significantly downregulated in cancerous tissues from 42 patients with EC who underwent surgical resection. Additionally, a low expression of miR­638 was significantly associated with advanced Federation of Gynecology and Obstetrics stage and was demonstrated to indicate shorter disease­free survival. Functional studies indicated that the overexpression of miR­638 in EC cell lines inhibited in vitro tumor progression and in vivo tumorigenicity. MEF2C was verified as a direct target of miR­638 and was demonstrated to mediate the tumor­suppressive function of miR­638 in EC.

Association between DNA damage repair gene somatic mutations and immune-related gene expression in ovarian cancer.

BACKGROUND: Defects in DNA damage repair (DDR) system may lead to genomic instability and manifest as increased immunogenicity. DDR deficiency is prevalent in ovarian cancer (OvCa); however, the association of DDR mutation with immune profiles in OvCa remains largely unknown. This knowledge will provide an essential basis to the rational design of biomarker-guided immune combination therapy of OvCa in the future. METHODS: Whole-exome sequencing data of 587 OvCa from The Cancer Genome Atlas (TCGA) were used to determine the expression profiles of 47 immune-related genes and the abundance of tumor-infiltrating immune cells. A Chinese OvCa cohort (n = 220) tested by next-generation sequencing (NGS) was used to validate the association between DDR status and tumor mutation burden (TMB). RESULTS: A total of 19.3% in TCGA cohort and 25.9% in Chinese cohort harbored at least one DDR somatic mutation. DDR deficiency exhibited a distinct immune profile with significant higher expression levels of PTPRCAP, CCL5, IFI16, LAG3, IL15RA, and GBP1 in OvCa in the TCGA cohort. Different DDR pathway deficiency displayed various immune profiles. Increased levels of Th1 cells, TMB, and neoantigen were also observed in DDR-deficient tumors. CONCLUSIONS: DDR deficiency was associated with specific immune signatures in OvCa. Our findings emphasize the urgent need for biomarker-guided rational immune combination therapy to maximize the OvCa patients who could benefit from immunotherapy.

Downregulation of eukaryotic initiation factor 4A1 improves radiosensitivity by delaying DNA double strand break repair in cervical cancer.

Expression of eukaryotic initiation factor 4A1 (eIF4A1) following brachytherapy has been reported to predict improved radiosensitivity and tumor-specific survival in cervical cancer. Therefore, the present study investigated the function of eIF4A1 in cervical cancer and the mechanism by which eIF4A1 regulates cervical cancer radiosensitivity. It was determined that the downregulation of eIF4A1 in HeLa and SiHa cells notably attenuated cell proliferation, in addition to repressing cervical cancer migration and invasion, and promoting cell apoptosis. In vitro and in vivo studies have demonstrated that silencing eIF4A1 improves cervical cancer radiosensitivity. Detection of γ-H2AX using western blot analysis at 0, 0.5, 1, 6 and 24 h following the exposure of cervical cancer cells to X-rays illustrated that eIF4A1-knockdown results in postponed radiation-induced DNA double strand break (DSB) repair. Overall, the results of the present study demonstrated that downregulated eIF4A1 improves cervical cancer radiosensitivity by delaying cancer cell DSB repair. In conclusion, the data indicated that eIF4A1 performs a vital role in cervical cancer progression and radiosensitivity. Therefore, eIF4A1 may be a potential therapeutic target in patients with cervical cancer.

Predictive factors of para-aortic lymph nodes metastasis in cervical cancer patients: a retrospective analysis based on 723 para-aortic lymphadenectomy cases.

Para-aortic lymph node (PALN) dissection is optional and controversial in patients with stage IB1-IIA2 cervical cancer. This retrospective study investigated PALN involvement patterns and evaluated preoperative clinical factors. A total of 723 consecutive FIGO stage IB1-IIA2 cervical cancer patients were included in the study. All patients underwent radical hysterectomy/radical trachelectomy, pelvic lymph node dissection, and PALN dissection. PALN metastasis was found in 101 (14.0%) patients, and the positive PALN rates of stage IB1, IB2, IIA1, and IIA2 were 8.4%, 11.1%, 17.2% and 21.7%, respectively. A multivariate model suggested age > 46 years (OR: 1.67, 95% confidence interval (CI): 1.08-2.58), tumor size > 3.5 cm (OR: 1.79, 95% CI: 1.12-2.87), and FIGO stage IIA (vs. IB) (OR: 1.97, 95% CI: 1.25-3.11) all positively correlated with PALN metastasis. When squamous cervical cancer cases were categorically analyzed, a multivariate model indicated age > 46 years (OR: 1.67, 95% CI: 1.00-2.80), FIGO stage IIA (vs. IB) (OR: 1.76, 95% CI: 1.02-3.02), and squamous cell carcinoma antigen (SCCA) > 6.5 ng/ml (OR: 5.20, 95% CI: 3.07-8.81) all positively correlated with PALN metastasis. Age, tumor size, and FIGO stage correlated with PALN metastasis in cervical cancer, while age, FIGO stage, and SCCA level were predictive in squamous cell carcinoma.

Diaphragmatic Surgery and Related Complications In Primary Cytoreduction for Advanced Ovarian, Tubal, and Peritoneal Carcinoma.

BACKGROUND: To evaluate the procedures and complications of diaphragm peritonectomy (DP) and diaphragm full-thickness resection (DFTR) during primary cytoreduction for advanced stage epithelial ovarian cancer. METHODS: All the patients with epithelial ovarian carcinoma who underwent diaphragm procedures at our institution between January 2009 and August 2015 were identified. Clinicopathological data were retrospectively collected from the patients' medical records. Postoperative morbidities were assessed according to the Memorial Sloan-Kettering Cancer Center (MSKCC) grading system. RESULTS: A total of 150 patients were included in the study. The majority of the patients had ovarian cancer (96%), stage IIIC disease (76%) and serous histology (89.3%). DP and DFTR were performed in 124 (82.7%) and 26 (17.3%) patients, respectively. A total of 142 upper abdominal procedures in addition to the diaphragmatic surgery were performed in 77 (51.3%) patients. No macroscopic residual disease was observed in 35.3% of the patients, while 84% of the total patient cohort had residual disease ≤1 cm. The overall incidence of at least one major morbidity (MSKCC grades 3-5) was 18.0%, whereas pleural effusions (33.3%), pneumonia (15.3%) and pneumothorax (7.3%) were the most commonly reported morbidities. The rate of postoperative pleural drainage was 14.6% in total, while half the patients in the DFTR group received drainage intraoperatively (11.5%) and postoperatively (38.5%). The incidence of postoperative pleural effusion was associated with stage IV disease (hazard ratio [HR], 17.2; 95% confidence interval [CI]: 4.5-66.7; P < 0.001), DFTR (HR, 4.9; 95% CI: 1.2-19.9; P = 0.028) and a long surgery time (HR, 15.4; 95% CI: 4.3-55.5; P < 0.001). CONCLUSIONS: Execution of DP and DFTR as part of an extensive upper abdominal procedure resulted in an acceptable morbidity rate. Pleural effusion, pneumonia and pneumothorax were the most common pulmonary morbidities. The pleural drainage rate was not high enough to justify prophylactic chest tube placement for all the patients. However, patients who underwent DFTR merited special consideration for intraoperative prophylactic drainage.

Extracellular matrix metalloproteinase inducer (CD147/BSG/EMMPRIN)-induced radioresistance in cervical cancer by regulating the percentage of the cells in the G2/m phase of the cell cycle and the repair of DNA Double-strand Breaks (DSBs).

Our preliminary study found that CD147 is related to radioresistance and maybe an adverse prognostic factor in cervical cancer. To date, the mechanisms underlying CD147-induced radioresistance in cervical cancer remain unclear. In the present study, we investigated the mechanisms by which CD147 affects radiosensitivity in cervical cancer both in vitro and in vivo. In this study, the clonogenic assay showed that radiosensitivity was significantly higher in the experimental group (the CD147-negative cell lines) than in the control group (the CD147-positive cell lines). After radiotherapy, the residual tumour volume was significantly lower in the experimental group. FCM analysis showed the cells percentage in the G2/M phase of the cell cycle were significantly higher in the CD147-negative group than in the control group. However, there was no significant difference in terms of apoptosis. The expression of gamma-H2A histone family, member X (γH2AX) was dramatically elevated in the CD147-negative cell lines after irradiation, but the expression of ataxia-telangiectasia mutated (ATM) was not different between the two groups. WB analysis did not show any other proteins relating to the expression of CD147. In conclusion, it is likely that CD147 regulates radioresistance by regulating the percentage of the cells in the G2/M phase of the cell cycle and the repair of DNA double-strand breaks (DSBs). Inhibition of CD147 expression enhances the radiosensitivity of cervical cancer cell lines and promotes post-radiotherapy xenograft tumour regression in nude mice. Therefore, CD147 may be used in individualized therapy against cervical cancer and is worth further exploration.

Decreased expression of EIF4A1 after preoperative brachytherapy predicts better tumor-specific survival in cervical cancer.

OBJECTIVE: The aim of this study is to investigate whether EIF4A1, EIF4E, and EIF4G1 can serve as prognostic markers for patients with cervical cancer receiving preoperative brachytherapy. MATERIALS AND METHODS: Tissue microarrays composed of 35 normal cervix samples, 87 cervical cancers treated without preoperative therapy, and 50 pairs of cervical cancer tissues collected before and after preoperative brachytherapy were constructed and evaluated for the expression of EIF4A1, EIF4E, and EIF4G using immunohistochemistry. Immunohistochemical staining was scored by the staining intensity and the percentages of tumor cells. The χ test was used to analyze the association between the immunohistochemistry results and clinicopathologic variables. The Kaplan-Meier method was applied to analyze the disease-specific survival. RESULTS: Overexpression of EIF4A1, EIF4E, and EIF4G1 were detected in 83.9%, 84.7%, and 80.3% of cervical cancers, respectively, all of which were significantly related to advanced International Federation of Gynecology and Obstetrics stage, squamous cell histology, lymph node metastasis, and deep stromal invasion (P < 0.05). The altered expression pattern of EIF4A1 and EIF4E after preoperative brachytherapy was significantly correlated with the cervical cancer response to brachytherapy (P = 0.029 and 0.012, respectively). The decreased expression of EIF4A1 predicted better tumor-specific survival (P = 0.02). The alteration of EIF4A1 was an independent predictor for tumor-specific survival (P = 0.047; hazards ratio, 0.272; 95% confidence interval, 0.076-0.982). CONCLUSIONS: Overexpression of EIF4A1, EIF4E, and EIF4G1 were acquired malignant phenotypic features of cervical cancer. EIF4A1 might function as a novel prognostic indicator and a potential therapeutic target for cervical cancer.

MicroRNA-181a enhances the chemoresistance of human cervical squamous cell carcinoma to cisplatin by targeting PRKCD.

MicroRNAs(miRNAs) are involved in regulating the response of cancer cells to various therapeutic interventions, but their involvement in the chemoresistance of human cervical squamous cell carcinoma is not fully understood. We found miR-181a was significantly up-regulated in specimens from patients with chemoresistant cervical squamous cell carcinoma. In this study, we aimed to clarify the role of miR-181a in regulating the chemoresistance of cervical cancer. Two human cervical squamous cancer cell lines, SiHa and Me180, were used. Enforced expression of miR-181a enhanced chemoresistance to cisplatin in cervical cancer cells through apoptosis reversion. In a nude mouse xenograft model, the overexpression of miR-181a markedly inhibited the therapeutic response to cisplatin. PRKCD, a target gene of miR-181a and a promoter of apoptosis, was negatively regulated by miR-181a. We found that the effect of miR-181a on chemoresistance was mediated by PRKCD. Additionally, silencing of PRKCD yielded an effect similar to that of miR-181a up-regulation and inhibited apoptosis in cervical cancer cells. Our findings suggest that miR-181a may function as an oncogene and induce chemoresistance in cervical squamous cell carcinoma cells at least in part by down-regulating PRKCD, thus may provide a biomarker for predicting chemosensitivity to cisplatin in patients with cervical squamous cancer.

Tiam1, negatively regulated by miR-22, miR-183 and miR-31, is involved in migration, invasion and viability of ovarian cancer cells.

Tiam1 has been implicated in the invasive phenotype of various carcinomas. However, its role in ovarian cancer remains to be elucidated, including its upstream regulatory mechanisms. In the present study, we examined the differential expression of Tiam1 in 10 normal ovarian tissues and 17 paired primary and corresponding metastatic ovarian cancer tissues by semi-quantitative immunohistochemistry. It was found that Tiam1 expression was remarkably increased in both primary and metastatic ovarian cancer tissues relative to normal ovarian tissues. Loss-of-function study revealed that downregulation of Tiam1 in SKOV-3ip and HO-8910PM cells lead to reduced cell migration and invasion, and growth inhibition without significantly affecting cell apoptosis. Subsequent regulatory study further confirmed the negative regulatory effects of miR-22, miR-183 and miR-31 on Tiam1 expression. Taken together, our data suggested that Tiam1 may be involved in the aggressive behavior of ovarian cancer, and differential expression profiles of microRNA (miRNA) may contribute to the dysregulation of Tiam1 abundance, which contributes to the invasive, migratory and viability properties of ovarian cancer cells.

Correlation between homeodomain-interacting protein kinase 2 and apoptosis in cervical cancer.

Homeodomain-interacting protein kinase 2 (HIPK2) is a serine/threonine nuclear kinase that is involved in apoptosis and cell growth, and is also thought to play a role in the process of tumorigenesis. The purpose of this study was to identify the role of HIPK2 in cervical cancer. HIPK2 expression was examined in normal and cervical cancer tissues at the mRNA and protein levels by quantitative real-time PCR and western blotting. To investigate the mechanism of action of HIPK2 in cervical cancer, RNA interference was used to analyze the effect of HIPK2 on apoptosis and cell growth in cervical cell lines. The results showed that HIPK2 expression was significantly higher in the cervical cancer tissues compared to the normal cervical tissues, both at the mRNA and protein level. Moreover, inhibition of HIPK2 promoted cell growth and decreased the rate of cell apoptosis in cervical cell lines. Taken together, these results indicate that HIPK2 expression is higher in cervical cancer tissues and has a positive correlation with cervical cancer. HIPK2 may be important in the development of cervical cancer.

[Potential role of ezrin and its related microRNA in ovarian cancer invasion and metastasis].

OBJECTIVE: to screen microRNA (miRNA) that inhibit expression of the metastasis-related gene ezrin in ovarian cancer cells and explore their correlation to the invasion and metastasis of ovarian cancer. METHODS: the differential expression of ezrin in two paired high-metastatic and low-metastatic cell lines were examined by real time reverse transcription (RT)-PCR and western blot. A functional screen with microarray was employed to identify miRNA that were differentially expressed between SKOV3 and SKOV3ip cell lines. Three programs, TARGETSCAN (http://www.targetscan.org), MICROCOSM (http://www.ebi.ac.uk/enright-srv/microcosm/htdocs/targets/v5/) and PICTAR (http://www.pictar.mdc-berlin.de), were employed to identify all miRNA, which may inhibit the expression of ezrin and were differentially expressed between SKOV3 and SKOV3ip cells. To test the repressive potential of these miRNA, synthetic mimetics were transfected individually into SKOV3ip cells and endogenous ezrin expression levels monitored by western blot and real-time RT-PCR. RESULTS: (1) the mRNA average level of ezrin were (81.74 ± 5.34)-fold higher expression level in SKOV3ip versus SKOV3 cells (P < 0.01), while (2.61 ± 0.14)-fold in HO-8910PM versus HO-8910 cells (P < 0.01). Elevated protein level of ezrin were observed in SKOV3ip cells compared with that in SKOV3 cells, and the same that in HO-8910PM cells compared with HO-8910 cells. Paired SKOV3 and SKOV3ip cells were employed to study the more significant difference in ezrin expression between them. (2) By a functional screen using miRNA microarray combined with bioinformatics analysis, the miR-183 and miR-22 were indentified as two candidate miRNA, which may have the potential regulatory role in ezrin expression. Real time RT-PCR assays revealed that miR-183 and miR-22 were, respectively, an average of (5.84 ± 0.66)-fold and (6.67 ± 0.67)-fold higher expression level in SKOV3ip versus SKOV3 cells (P < 0.01), which were in agreement with the microarray data. A subsequent validation by western blot and real time RT-PCR revealed that over-expression of miR-183 and miR-22 could both lead to an obvious decrease in ezrin protein level, while there were not signicant difference in the level of ezrin mRNA (P > 0.05). CONCLUSION: increased expression of miR-183 and miR-22 may both repress the protein level of ezrin, indicating that miR-183 and miR-22 may bear a potential role in inhibiting ovarian cancer metastasis in a ezrin-mediated way.

[Screening and identification of potential miRNA involved in ovarian cancer invasion and metastasis].

OBJECTIVE: To identify potential miRNA involved in epithelial ovarian cancer (EOC) invasion and metastasis. METHODS: miRNA microarray was applied to compare the miRNA expression profile between SKOV-3ip and SKOV-3 cells. Bioinformatics programs (TargetScan, MicroCosm, PicTar and GO) were used to analyze the miRNA and their potential target genes. Real-time RT-PCR was used to confirm the results of microarray and for expanding detection in another paired EOC cell lines (HO-8910 and HO-8910PM). RESULTS: Totally, expressions of 42 miRNA were found significantly different between SKOV-3ip and SKOV-3 cells. Among them, 10 miRNA were down-regulated, including let-7a, let-7f, miR-22 and miR-886-5p; while 32 were up-regulated, for example, let-7e and miR-519e. Bioinformatic analysis indicated that let-7a, let-7e, let-7f, miR-22 and miR-886-5p may be involved in cancer invasion and metastasis. Meanwhile, real-time RT-PCR confirmation and statistic analysis showed that let-7f and miR-22 expressions were significantly different between ovarian cancer cell lines with various invasive and metastatic capacity (P < 0.05). CONCLUSION: The expression of let-7f and miR-22 is low in ovarian cancer cells with high invasive and metastatic capacity. It suggests that they are potential tumor suppressor genes. Further research on their role and mechanism is needed.

An inhibitory effect of miR-22 on cell migration and invasion in ovarian cancer.

OBJECTIVES: Aberrant expression of microRNAs (miRNAs) has been implicated in ovarian carcinoma. However, roles of miRNAs in ovarian caner metastasis have not been comprehensively addressed. This work is aimed to identify selected miRNAs involved in ovarian cancer metastasis. METHODS: We examined the distinct miRNA expression profiles between paired high-metastatic human serous ovarian cancer cell SKOV-3ip and low-metastatic human serous ovarian cell SKOV-3 using miRNA microarray. Subsequently, a validation with Real-time RT-PCR was performed for miR-22 expression level, and a functional study was carried out for miR-22. RESULTS: Through a screen with microarray, we found there were a variety of miRNAs differentially expressed between paired high and low metastatic serous ovarian cancer cells. Particularly, miR-22 was identified as a potential metastasis-inhibitor in ovarian cancer. There was a negative correlation between miR-22 expression and the metastatic potential in ovarian cancer cells. Furthermore, both gain-of-function and loss-of-function studies displayed an inhibitory effect of miR-22 on cell migration and invasion in vitro without significantly affecting cell viability and apoptosis. Subsequent bioinformatics analysis revealed that miR-22 might regulate multiple pro-metastatic genes, which could provide an explanation to the inhibitory effects of miR-22 on cell migration and invasion. CONCLUSIONS: Taken together, our findings suggested that miR-22 might be involved in inhibiting ovarian cancer metastasis.