Exosomal circNFIX promotes angiogenesis in ovarian cancer via miR-518a-3p/TRIM44 axis.

Ovarian cancer (OC) is a gynecological cancer with high mortality. OC-derived exosomal circRNAs can regulate angiogenesis. This study aims to explore the role and mechanism of exosomal circRNA nuclear factor I X (CircNFIX) derived from OC cells in angiogenesis. Quantitative real-time polymerase chain reaction was employed to evaluate the levels of circNFIX, miR-518a-3p, and tripartite motif protein 44 (TRIM44) in OC and adjacent tissues. Exosomes from the ovarian surface epithelial cell (HOSEpiC) and OC cells (SKOV3 or OVCAR3) were isolated by differential centrifugation. Exosomes were cocultured with the human umbilical vein endothelial cells (HUVECs). The angiogenesis capacity was analyzed by Tube formation assay. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and Transwell assays were used to determine the cell viability and migration ability. The dual-luciferase report, RNA immunoprecipitation (RIP), and RNA pull-down assays were applied to validate the gene's interaction. CircNFIX and TRIM44 expression were higher and miR-518a-3p was lower in OC tissues than in the adjacent tissues. Upregulated circNFIX and TRIM44 were significantly correlated with the tumor size and International Federation of Gynecology and Obstetrics (FIGO) stage of OC patients. HUVECs treated OC-derived exosomes had higher proliferation, migration, and angiogenesis capacities than the control group. While OC-derived exosomal circNFIX silencing restrained HUVECs' proliferation, migration, and angiogenesis, compared with the OC-derived exosomes group. OC-derived exosomal circNFIX positively regulated TRIM44 expression by targeting miR-518a-3p in HUVECs. OC-derived exosomal circNFIX promoted angiogenesis by regulating the Janus-activated kinase/signal transducer and activator of transcription 1 (JAK/STAT1) pathway via miR-518a-3p/TRIM44 axis in HUVECs.

CircNFIX stimulates the proliferation, invasion, and stemness properties of ovarian cancer cells by enhancing SH3RF3 mRNA stability via binding LIN28B.

We aimed to study the regulatory roles and mechanism of circular nuclear factor IX (circNFIX) in cancer growth and stemness properties of ovarian cancer (OC). CircNFIX and SH3RF3 levels in OC tissues and cells were tested by quantitative real-time PCR. RNase R treatment quantified circNFIX RNA stability. Molecular interaction among circNFIX, LIN28B, and SH3RF3 was predicted by bioinformatics software and validated through RNA immunoprecipitation (RIP) assay. The gain- or loss-experiments of circNFIX on capabilities of metastasis and stemness in vitro were assessed using Cell Counting Kit-8, Transwell, western blot, and sphere-formation assays. CircNFIX and SH3RF3 were markedly elevated in OC tissues and OC cells. Knocking down circNFIX repressed the proliferation, migration, invasion, and stemness properties of A2780 and SKOV3 cells. The RIP assay verified the direct binding relationship between LIN28B, circNFIX, and SH3RF3. Additionally, overexpression of circNFIX elevated the SH3RF3 expression, while this effect was reversed by LIN28B silence. Rescue experiments demonstrated that the overexpression of SH3RF3 reversed the knockdown of circNFIX on OC cells' proliferation, metastasis, and stemness properties. CircNFIX improved the mRNA stability and translation of SH3RF3 via recruiting LIN28B, thus promoting the proliferation, invasion, and stemness properties of OC cells in vitro.

TRIM44 facilitates ovarian cancer proliferation, migration, and invasion by inhibiting FRK.

Ovarian cancer (OC) is the leading cause of gynecologic cancer-related death in the world. Accumulating evidence indicated the important role of TRIM44 in cancer development. However, how TRIM44 displays in OC and the underlying mechanism remained unclear. TRIM44 and FRK expression in OC tissues and cell lines were investigated by western blot and RT-qPCR. Histotype of tissue samples and patients' data were analyzed. Kaplan-Meier Curve was performed to validate the effect of TRIM44. Colony formation assay, MTT assay, Transwell assay, and wound-healing assay were applied to elucidate the function of TRIM44 in OC cells. CHIP assay was used to explore the association between TRIM44 and FRK. Finally, we performed SKOV3 xenografts in Balb/c nude mice to further confirm the involvement of TRIM44 in OC development. We found TRIM44 highly expressed while FRK displayed low expression in OC cell lines and tissues. Moreover, analysis of histotype of tissues and patients' data and Kaplan-Meier Curve implied the important role of TRIM44 and FRK in tumor progression. Further in vitro study suggested that knocking down TRIM44 inhibited OC cells proliferation, migration, and invasion. Besides, FRK was identified as the target gene of TRIM44 in OC, and TRIM44 promoted OC cells proliferation, migration, and invasion by inhibiting FRK. Finally, in vivo animal experiment further confirmed the promotive effect of TRIM44 on OC progression. Our findings demonstrated that TRIM44 facilitated OC proliferation, migration, and invasion by inhibiting FRK, providing new insights for theoretical research and therapy of OC.

[Clear Cell Carcinoma of Cervix in a 12-Year-Old Girl: A Case Report].

The 12-year-old patient was admitted to the hospital on September 19, 2019 for "vaginal bleeding for 2 + months and pelvic mass to be diagnosed". The patient and her family explicitly denied any previous history of diethylstilbestrol exposure. After admission, relevant examinations were conducted and hysteroscopic exploration was performed under general anesthesia. During the procedure, cervical neoplasms were extracted and pathology results indicated cervical cancer. Then, extensive transabdominal hysterectomy+bilateral salpingectomy+bilateral ovarian transposition+pelvic lymph node dissection+para-aortic lymph node sampling were performed. Postoperative pathology analysis of the removed tissue showed that clear cell carcinoma of cervix (CCAC) had infiltrated into 1/3 of the cervical stroma and there was downward involvement of the vaginal wall; the cancer metastasized to the left obturator lymph node and the left internal and external iliac lymph nodes. Immunohistochemical staining of the removed tissue showed the following results: cytokeratin 7 (+), cytokeratin 20 (-), Napsin-A (+), cell adhesion molecule CD15 (+), heatocyte nuclear factor-1 ß (+), Sal-like protein 4 (-), tumor suppressor gene P16 protein (+), estrogen receptor (+), progesterone receptor (-), tumor suppressor gene P53 protein (focal positive), tumor suppressor gene WT-1 protein (-) and Ki67 antigen (about 40% positive). The patient was diagnosed with CCAC stage ⅢC1p. Four cycles of postoperative systemic chemotherapy (fluorouracil+cisplatin) and 25 times of three-dimensional afterloading radiotherapy were performed. The patient did follow-up visits and did not show obvious signs of recurrence. The clinical manifestations of this disease are basically the same as those of cervical squamous cell carcinoma, and if the patient is younger, it can be easily misdiagnosed as dysfunctional uterine bleeding, indicating the need for differential diagnosis.

Postmolar choriocarcinoma after an interval of 7 years: Case report and literature review.

Choriocarcinoma is a rare pregnancy-related malignancy. The majority is arising from non-molar pregnancy. Here we report a patient who was diagnosed with postmolar choriocarcinoma after an interval of 7 years. Before surgery, we suspected the diagnosis of the patient was intramural pregnancy or choriocarcinoma. Laparoscopy was performed and hysterectomy was carried out. Postoperative pathological evaluation of the surgical specimen confirmed the case was choriocarcinoma. Hysterectomy through laparoscopy was feasible and safe for selected patients.

[Pathological characteristics and prognosis of 664 patients with epithelial ovarian cancer: a retrospective analysis].

OBJECTIVE: To analyze clinicopathologic features and risk factors associated with the recurrence and prognosis of epithelial ovarian cancer (EOC). METHODS: 710 EOC patients treated at the West China University Second Hospital from Jan. 2006 to Jun. 2011 were recruited in this study retrospectively. Univariate and multivariate logistic regression models were constructed to evaluate the risk of factors. Kaplan-Meier and log-rank methods were adopted for survival analyses. RESULTS: The final sample included 664 patients with complete and well-documented data. The participants had a mean age of (49.35 +/- 11.58) yr. and 79.07% (525/664) were older than 40 year-old. CA125 was tested in 550 patients before surgery and 82.55% showed abnormal values. Over half (55.57%) of the patients were classified as serious EOC, which was followed by clear cell EOC (12.35%), endometrioid EOC (10. 09%), mucinous EOC (7.68%), and others (14.31%). Stage I (FIGO) comprised 30. 72% of the cases. The patients were followed up on average of (37.48 +/- 12.51) months and 303 died with a mean survival length of (28.54 +/- 9.56) months. Recurrence was found in 126 patients at a median of (26.10 +/- 5.75) months. For the 361 survived, 81. 72% lived without detectable cancer. All patients received surgical operations, including 483 undergoing retroperitoneal lymphadenectomy. The univariate analysis identified older age, advanced FIGO stage, suboptimal debulking, abnormal CA125 values before surgery, undifferentiated, mixed type and serous pathologic subtypes, poor-differentiation and pathogenesis of tumor as risk factors associated with survival prospect. The multivariate logistic regression models confirmed that poor prognosis was associated with older age, advanced FIGO stage, suboptimal debulking and undifferentiated, mixed type and serous pathologic subtype. CONCLUSION: Older age, advanced FIGO stage, high grade differentiation, suboptimal debulking, lymphnode metastasis, and type II EOC are associated with poor prognosis of EOC patients.

[Expression of AIB1 protein in epithelial ovarian cancer cells and its impact on apoptosis].

OBJECTIVE: To investigate the expression of AIB1 (amplified in breast cancer 1) protein in epithelial ovarian cancer cells and to analyze the relationship between AIB1 protein and the apoptosis modulated proteins: P53 and Bcl-2. METHODS: The expressions of AIB1, P53 and Bcl-2 proteins in 24 normal ovaries, 24 benign ovarian tumors, 18 borderline ovarian tumors and 69 epithelial ovarian cancers were examined with immunohistochemical method SP. The expressions of estrogen receptor (ER) and progesterone receptor (PR) were also examined to reveal their relation with AIB1 expression. RESULTS: 1) The AIB1 protein expressed in 65.22% ovarian cancers, greater than in normal ovaries (8.33%), benign ovarian tumors (25.00%) and borderline ovarian tumors (38.89%). The overexpression of AIB1 protein occurred in 36.23% ovarian cancers. Ovarian cancers poorly-differentiated, at a late clinical stage and with lymph node involvement had higher AIB1 overexpression than those well-differentiated and moderately-differentiated, and those at an early clinical stage and without lymph node involvement. 2) The expression of AIB1 was positively correlated with the expression of P53 (r = 0.342, P = 0.004) and Bcl-2 proteins (r = 0.311, P = 0.009) in the ovarian cancers. There was no correlations between the AIB1 expression and the ER and PR. CONCLUSION: AIB1 protein expression increases in ovarian cancers, which is associated with the higher malignant biological behavior. The overexpression of AIB1 may be involved in the carcinogenesis and the development of epithelial ovarian cancers through a hormone independent pathway. Our findings suggest a possible correlation between AIB1 and apoptosis modulated proteins P53 and Bcl-2.

[P53 codon 72 polymorphism in cervical cancers and its correlation with HPV16,18E6].

OBJECTIVES: To examine p53 codon 72 polymorphism in cervical cancers and its correlation with HPV16/18 E6. METHODS: Cervical specimens were taken from 81 patients with cervical squamous cancer, 18 patients with cervical adenocarcinoma, 88 patients with CIN II - III and 60 patients without cancers. PCR was used to examine the p53 genotypes and the expression of HPV16 and 18 E6. RESULTS: The frequencies of p53 Arg homozygosity in cervical squamous cancer, cervical adenocarcinoma and CIN (II - III) were 58.020%, 55.55% and 59.09% respectively, greater than those of p53 Arg/Pro heterozygosity (30.86%, 27.78%, 21.59%) and those of p53 Pro homozygosity (11.12%, 16.67%, 19.32%). The normal cervical samples also showed less frequency of p53 Arg homozygosity (23.33%) than cervical squamous cancer. There were no significant differences in the frequencies of p53 Arg homozygosity, p53 Arg/Pro heterozygosity and p53 Pro homozygosity (23.33%, 40.00% and 36.67% respectively). The frequency of HPV16,18 E6-positive in cervical cancer and CIN was much higher than that in control group (81.84%, 50.00% and 53.41%) for the normal cervical samples. The expression of HPV16 and 18 E6 in cervical squamous cancers was more frequent than in CIN. The frequency of p53 Arg homozygosity in HRHPV E6-positive cervical squamous cancers (64.06%) was greater than in HRHPV E6-negative cervical squamous cancers (35.29%) and in HRHPV E6-positive normal samples (33.33%). The p53 codon 72 polymorphism showed no differences in samples with different FIGO staging and grades. CONCLUSION: p53 Arg homozygosity could serve as a risk indicator for the tumorigenesis of cervix. In combination with HRHPV E6, it might be able to predict the progression of cervical lesions. p53 codon 72 polymorphism is not associated with FIGO staging and grades of cervical cancers.

[Evaluation of the effect of fertility-saving surgery on young patients with malignant ovarian tumors].

OBJECTIVE: To evaluate the impact of fertility-saving surgery and adjuvant chemotherapy on survival and fertility of young patients with ovarian malignant tumors. METHODS: A retrospective analysis was done on 38 patients with ovarian malignant germ cell tumors, 22 patients with malignant epithelial tumors and 4 patients with sexual cord mesenchymal tumors receiving conservative treatments. Outcomes such as menstruation and reproduction ability were assessed. RESULTS: Fifty-nine among 64 patients have been alive up to now (92%). The overall survival rate for ovarian epithelial malignancies, malignant germ cell tumors and sexual cord mesenchymal tumors were 95% (21/22), 89% (34/38) and 4/4, respectively. Fifteen patients received second operation and recurrence was found in 6 patients. Among the 59 surviving patients, 53 had normal menstruation. Thirteen patients among 20 patients who wanted to get pregnant had 15 pregnancies and 9 successful deliveries. CONCLUSIONS: The management of fertility-saving surgery on patients with ovarian malignant germ cell tumors, whatever the stagings are, is a safe option. For patients with ovarian epithelial carcinomas, fertility-saving surgery is only indicated for low-stage (stage I), high-grade (G1), and patients who hope to maintain fertility function eagerly. Cisplatinum-based combination chemotherapy is necessary. Standardized chemotherapy has no effect on fertility function.