Identification of a ferroptosis-related long non-coding RNA signature for prognosis prediction of ovarian cancer.

Ovarian cancer is one of the deadliest malignant tumors. Ferroptosis is closely related to various cancers, including ovarian cancer, but the genes involved in regulating ferroptosis in ovarian cancer are still unclear. The aim of this study is to construct a long non-coding RNA (lncRNA) signature related to ferroptosis and evaluate its relationship with the prognosis and clinicopathological characteristics of patients with ovarian cancer. In this study, a prognostic risk model comprising 18 lncRNAs related to ferroptosis was obtained. Compared to the low-risk group, the high-risk group based on the FerRLSig score had significantly poorer overall survival (P < 0.001). The receiver operating characteristics curve supported the accuracy of the model, established a prognostic nomogram combining FerRLSig and clinical characteristics, and showed a good prognosis and survival risk stratification predictive power. In addition, Gene Set Enrichment Analysis (GSEA) showed that FerRLSig was involved in many malignant tumor-related immunomodulatory pathways. Based on the risk model, we found that immune status and immunotherapy, chemotherapy, and targeted therapy were significantly different between the high-risk and low-risk groups. This study provided a more in-depth understanding of the molecular and signaling pathways of ferroptosis in ovarian cancer and showed the impact of tumor microenvironment on ovarian cancer, as well as provided a prognostic model for ovarian cancer patients to guide the clinical treatment of ovarian cancer.

Wnt-11 overexpression promoting the invasion of cervical cancer cells.

Wnt-11 is a positive regulator of the Wnt signaling pathway, which plays a crucial role in carcinogenesis. However, Wnt-11 expression in cervical cancer has not been well investigated. The aim of this study was to investigate the role of Wnt-11 in cervical tumor proliferation and invasion. This study examined 24 normal cervical squamous epithelia, 29 cervical intraepithelial neoplasia (CIN), and 78 cervical cancer samples. The expression of Wnt-11 was investigated by immunohistochemistry and quantitative reverse transcription-polymerase chain reaction analysis. The expression of the high-risk human papilloma virus (HR-HPV) E6 oncoprotein was also investigated by immunohistochemistry. In addition, the expression of Wnt-11, HR-HPV E6, JNK-1, phosphorylated JNK-1(P-JNK1), and ß-catenin was examined by western blot analysis following Wnt-11 knockdown or overexpression in HeLa or SiHa cells, respectively. The promotion of cervical cancer cell proliferation and invasion was investigated using the cell counting kit-8 and Matrigel invasion assay, respectively. Wnt-11 and HR-HPV E6 expression increased in a manner that corresponded with the progression of cervical cancer and was significantly correlated with the International Federation of Gynecology and Obstetrics cancer stage, lymph node metastasis, tumor size, and HPV infection. Wnt-11 protein expression was positively associated with HR-HPV E6 protein expression in all 78 cervical cancer samples (P < 0.001). Furthermore, Wnt-11 was positively associated with P-JNK1 expression and promoted cervical cancer cell proliferation and invasion. These observations suggest that the increased Wnt-11 expression observed in cervical cancer cells may lead to the phosphorylation and activation of JNK-1 and significantly promote tumor cell proliferation and cell migration/invasion through activation of the Wnt/JNK pathway. Consequently, Wnt-11 may serve as a novel target for cervical cancer therapy.

Sex-determining region Y-related high mobility group box (SOX)-2 is overexpressed in cervical squamous cell carcinoma and contributes cervical cancer cell migration and invasion in vitro.

Sex-determining region Y-related high mobility group box 2 (SOX-2) is a key pluripotency-associated transcription factor and may be implicated in the pathogenesis of cervical squamous cell carcinoma (SCC). The aim of this study was to explore SOX-2 expression in cervical SCC tissues and to examine whether and how SOX-2 regulates the malignant behaviors of cervical SCC cells in vitro. We here found that SOX-2 expression in the examined cervical SCC tissues was higher than that in the normal cervical and cervical intraepithelial neoplasia (CIN) tissues. Higher protein level of SOX-2 (nuclear positive staining cells ≥50 %) was detected in 34.9 % (29 out of 83 cases) of cervical SCC patients. We also noted that 100 % of well-differentiated and 66.7 % of moderately differentiated cervical SCCs showed lower SOX-2 expression (nuclear positive staining cells <50 %), while 58.8 % of poorly differentiated tumors had higher SOX-2 expression (P < 0.05). Furthermore, the migratory and invasive capabilities of SiHa cervical cancer cells were enhanced when SOX-2 was upregulated whereas suppressed when SOX-2 was downregulated. Also, the phosphorylation levels of protein kinase B (Akt) and extracellular regulated protein kinases (ERK) 1/2 were increased in SOX-2-overexpressed cancer cells but decreased in SOX-2-depleted cells. Additionally, LY294002 (Akt pathway inhibitor) or U0126 (ERK pathway inhibitor) significantly suppressed SOX-2-overexpression-induced migration and invasion in SiHa cells. Our results indicate that differentially expressed SOX-2 is associated with tumor differentiation (P < 0.05) and that SOX-2 contributes to the migratory and invasive behaviors of cervical SCC in vitro.

[Expression of Daxx and HPV16 E6 and its significance in cervical lesions].

OBJECTIVE: To investigate the expression of death domain associated protein (Daxx) and human papillomavirus 16 E6 protein (HPV16 E6) in cervical lesions and analyze their significance. METHODS: Immunohistochemical SABC method was used to detect the expression of Daxx and HPV16 E6 in 194 cases of cervical tissues with different lesions. RESULTS: (1) The positive expression rate of Daxx was 28.57% (12/42), 40.00% (18/45), 65.91% (29/44), 66.67% (42/63), respectively, in chronic cervicitis, cervical intraepithelial neoplasia I-II (CIN I-II), CIN III and cervical squamous cell carcinoma. The positive expression rate of HPV16 E6 was 15.38% (6/39), 36.17% (17/47), 46.30% (25/54), 100.00% (24/24) in the above four groups. The positive rates in cervical cancer group and high grade CIN group were significantly higher than these in low level of CIN group and chronic cervicitis group (P<0.05). (2) The expression of Daxx was stronger in HPV16 E6 high positive group than that in HPV16 E6 low positive group (P<0.05). (3) There was no significant relationship between the expression of Daxx and pathological classification, clinical stage, situation of lymph node metastasis or patients' age in cervical squamous cell carcinoma (P>0.05). (4) There was significant positive relationship between Daxx and HPV16 E6 expression (r=0.695, P<0.05). CONCLUSIONS: The expression of Daxx and HPV16 E6 gradually increases with cervical lesion degree aggravating. Here might be synergy between them, and both could promote the development of cervical lesions.

Expression and effects of high-mobility group box 1 in cervical cancer.

We investigated the significance of high- mobility group box1 (HMGB1) and T-cell-mediated immunity and prognostic value in cervical cancer. HMGB1, forkhead/winged helix transcription factor p3 (Foxp3), IL-2, and IL-10 protein expression was analyzed in 100 cervical tissue samples including cervical cancer, cervical intraepithelial neoplasia (CIN), and healthy control samples using immunohistochemistry. Serum squamous cell carcinoma antigen (SCC-Ag) was immunoradiometrically measured in 32 serum samples from 37 cases of squamous cervical cancer. HMGB1 and SCC-Ag were then correlated to clinicopathological characteristics. HMGB1 expression tends to increase as cervical cancer progresses and it was found to be significantly correlated to FIGO stage and lymph node metastasis. These findings suggest that HMGB1 may be a useful prognostic indicator of cervical carcinoma. In addition, there were significant positive relationships between HMGB1 and FOXP3 or IL-10 expression (both p<0.05). In contrast, HMGB1 and IL-2 expression was negatively correlated (p<0.05). HMGB1 expression may activate Tregs or facilitate Th2 polarization to promote immune evasion of cervical cancer. Elevated HMGB1 protein in cervical carcinoma samples was associated with a high recurrence of HPV infection in univariate analysis (p<0.05). HMGB1 expression and levels of SCC-Ag were directly correlated in SCC (p<0.05). Thus, HMGB1 may be a useful biomarker for patient prognosis and cervical cancer prediction and treatment.

Coordination of Single-cell and Bulk RNA Sequencing to Construct a Cuproptosis-related Gene Prognostic Index for Endometrial Cancer Prognosis, Immune Microenvironment Infiltration, and Immunotherapy Treatment Options.

Background: This study aims to identify molecular subtypes and develop a cuproptosis-related gene prognostic index (CRGPI) for endometrial cancer (EC), in addition to outlining the immune features and the efficacy of immune checkpoint inhibitor (ICI) therapy in CRGPI-defined groups of EC. Methods: Between malignant and normal cells distinguished from single-cell RNA sequencing data GSE154763 dataset, the difference in KEGG pathways and cuproptosis-related gene (CRG) scores was intensively investigated. On the basis of TCGA dataset (n=492), CRGs were used to identify two distinct molecular subtypes. Using the Cox regression method, a CRGPI was constructed and externally validated with the IMvigor210 dataset (n=348) and GSE78220. Then, the molecular and immune characteristics and the efficacy of ICI therapy in subgroups defined by CRGPI were investigated. Results: Compared to normal cells, the expression of the TCA cycle and CRGs genes was significantly higher in malignant cells. The CRGPI was established on the premise of ATF5, FBXO46, P2RX4, SMARCD3, DAPK3, and C1orf53. Comprehensive results demonstrated a correlation between a low CRGPI score and better overall survival, younger age, early stage, immune cells and functions activation, high tumor mutation burden and high microsatellite instability, as well as better benefit from ICI therapy, and its significance for forecasting immunotherapeutic effects was verified as well (IMvigor210 cohort: HR, 1.358; 95% CI, 1.047, 1.761; p=0.02; GSE78220 cohort: HR, HR = 3.857, 95% CI, 1.009, 14.74; p=0.034). CRGPI anticipated the immunotherapy medication. Conclusions: CRGPI is a prospective biomarker to estimate the prognosis, immune and molecular characteristics, and treatment benefit of immunotherapy in EC.

Uterine incision dehiscence 3 mo after cesarean section causing massive bleeding: A case report.

BACKGROUND: The traditional definition of late postpartum hemorrhage is a massive uterine hemorrhage from 24 h after delivery to the puerperal period. Here, we report a case of late postpartum hemorrhage that occurred 3 mo after cesarean section and endangered the patient's life. The cause of the case we are reporting was poor incision healing. By reporting this case, we hope to make doctors aware that late postpartum hemorrhage due to poor incision healing may happen as late as 3 mo after cesarean section. CASE SUMMARY: A 31-year-old woman complained of acute, severe vaginal bleeding for 1 h; the patient had a history of cesarean section 3 mo prior. After receiving anti-inflammatory treatment, fluid supplementation, blood transfusion, oxytocin administration, and hemostatic treatment, the vaginal bleeding ceased, and the patient's clinical status improved. Unfortunately, she experienced recurrent massive vaginal bleeding, and uterine contractile agents did not decrease the persistent bleeding. To save the patient's life, she was admitted for emergency laparotomy. At exploratory laparotomy, dehiscence and necrosis of the previous cesarean section scar were noted; the dehiscence penetrated through the entire thickness of the uterine muscle wall and extended to the left uterine artery. Ultimately, we performed a total hysterectomy. CONCLUSION: Late postpartum hemorrhage due to poor incision healing after cesarean section may occur in the 3 mo after cesarean section or even later. Therefore, obstetricians-gynecologists should monitor for this potential complication in all patients post-cesarean section. Such hemorrhages can be severe enough to endanger the patient's life.

Circular RNA circNRIP1 promotes migration and invasion in cervical cancer by sponging miR-629-3p and regulating the PTP4A1/ERK1/2 pathway.

Emerging evidence indicates that circRNAs play essential roles in tumorigenesis and development. However, the role of circRNAs in cervical cancer (CC) remains unclear. CircRNA microarrays performed on the immortal cervical cell line H8 and the cervical cancer cell line SiHa were used to identify a circRNA, termed circNRIP1 (hsa_circ_0004771), which was upregulated in SiHa. QRT-PCR confirmed that circNRIP1 was upregulated in CC tissues, where its expression was correlated with lymphovascular space invasion. Besides, both in vitro and in vivo experiments demonstrated that circNRIP1 promotes cell proliferation, migration, and invasion. Additionally, we found that miR-629-3p induced tumor suppression by regulating PTP4A1 and the ERK1/2 pathway. Finally, we confirmed that circNRIP1 exerts its effect, at least partially, by sponging miR-629-3p and thereby regulating the PTP4A1/ERK1/2 pathway. Therefore, circNRIP1 may be useful as a potential prognostic biomarker and therapeutic target in CC patients.

High-mobility group box 1 is overexpressed in cervical carcinoma and promotes cell invasion and migration in vitro.

The present study aimed to investigate the expression of high-mobility group box 1 protein (HMGB1) in cervical carcinoma and explore whether or not HMGB1 promotes cervical carcinoma cell invasion and migration in vitro and the related mechanism. HMGB1, nuclear factor-κB (NF-κB), E-cadherin and N-cadherin protein expression was analyzed in tissues from 48 cervical carcinomas, 51 cervical intraepithelial neoplasia (CIN) and tissues from 24 healthy controls using immunohistochemistry. HeLa cells were treated with different concentrations of HMGB1 (0, 10, 100, and 1,000 ng/ml) at different time-points (0, 24, 48 and 72 h), and changes in cell morphology and biological behaviors were observed. Changes in the expression levels of E-cadherin, N-cadherin, NF-κB and the inhibitor κB (IκB) in the treated cells were detected by western blot analysis and real-time PCR. HMGB1 expression exhibited a gradually increasing trend in the normal cervical tissues, CIN and cervical cancer, and there was statistical significance between the three groups (P<0.05). HMGB1 expression level was associated with FIGO stage, lymph node metastasis and differentiation (P<0.05). HMGB1 expression was positively related to N-cadherin and NF-κB; and HMGB1 had a negative relationship with E-cadherin. HMGB1 stimulation caused HeLa cells to lose cell polarity and transition from epithelial cells into spindle-shaped cells with sparse cell-cell junctions. The expression levels of E-cadherin and IκB in the cytoplasm were reduced, while N-cadherin expression was increased. The level of NF-κB expression in the nucleus was also increased. Treatment with NF-κB inhibitor (BAY11-7082) and receptor for advanced glycation end products (RAGE) antagonist (anti-RAGE) significantly suppressed HMGB1­mediated epithelial-to-mesenchymal transition in the HeLa cervical cancer cells. The results suggest that HMGB1 is associated with outcomes of cervical cancer and promotes subsequent invasion and metastasis of cervical cancer cells by activating the NF-κB signaling pathway. This potential mechanism could be an important determinant of cervical cancer metastasis.

B7-H3, B7-H4, Foxp3 and IL-2 expression in cervical cancer: Associations with patient outcome and clinical significance.

The aim of this study was to determine the expression of B7-H3, B7-H4, Foxp3 and IL-2 in cervical cancer tissues, and evaluate the corresponding clinical significance. The expression of B7-H3, B7-H4, Foxp3 and IL-2 in 108 cervical cancer specimens was detected using immunohistochemistry, and their relationship with clinicopathologic parameters was determined. B7-H3, B7-H4 and Foxp3 had high levels of expression in cervical cancer cells (72.22, 80.56, and 91.56%, respectively). B7-H3 levels were only significantly associated with tumor size (P=0.013), while B7-H4, Foxp3 and IL-2 levels were significantly associated with International Federation of Gynecology and Obstetrics (FIGO) stage (P=0.023, 0.014 and 0.036, respectively) and tumor size (P=0.045, 0.010 and 0.021, respectively). Their expression levels were not correlated with age, histologic type, differentiation and lymph node metastasis (all P>0.05). Cox regression multivariate analysis confirmed that B7-H3 or B7-H4 overexpression was an independent prognostic factor. In addition, there were significant positive relationships between the expression of B7-H3 and B7-H4 with Foxp3 (P<0.001). In contrast, the expression of B7-H3 and B7-H4 was negatively correlated with IL-2 (P<0.05). B7-H3, B7-H4 and Foxp3 may be useful biomarkers in patients with cervical cancer for predicting treatment.

Roles of Foxp3 in the occurrence and development of cervical cancer.

OBJECTIVE: This study aimed to evaluate the relationship between forkhead box P3 (Foxp3) expression and clinicopathological characteristics of cervical cancer and to explore the influence of Foxp3 on the biological behaviors of cervical cancer cells. METHODS: In this study, immunohistochemistry, lentivirus mediated transfection, Transwell assay; CCK-8 assay, real-time PCR and flow cytometry were employed to confirm the roles of Foxp3 in the occurrence and development of cervical cancer. RESULTS: Foxp3 and p16(INK4a) were highly expressed in the cervical cancer and their expressions were related to the FIGO stage, tumor size, lymph node metastasis and serum SCC. Foxp3 had a high expression in the cervical cancer cells, tumor interstitium and metastatic lymph nodes. Foxp3 expression was positively related to p16(INK4a) expression in the cervical cancer. Foxp3 expression in the cervical cancer was negatively related to the prognosis: high Foxp3 expression predicted a poor prognosis. Silencing of Foxp3 was able to inhibit the proliferation and invasiveness of cervical cancer cells, promote their apoptosis, and induce the change in cell cycle. Silencing of Foxp3 also reduced the mRNA and protein expressions of p16(INK4a) in cervical cancer cells. CONCLUSION: Foxp3 is highly expressed in the cervical cancer, and able to facilitate the proliferation and invasiveness of cervical cancer, change cell cycle and inhibit their apoptosis, resulting in the occurrence, development and metastasis of cervical cancer.

Clinical significance of Wnt-11 and squamous cell carcinoma antigen expression in cervical cancer.

The purpose of the study was to determine the expression patterns of Wnt-11 and squamous cell carcinoma antigen in cervical cancer tissues and to explore their clinical significance and correlation with clinicopathological parameters. The expression of Wnt-11 and squamous cell carcinoma antigen was detected in 127 cervical cancer tissues, 21 cervical intraepithelial neoplasia, as well as in 20 healthy controls by immunohistochemistry, and the relationship of Wnt-11 and squamous cell carcinoma antigen expression with clinicopathological parameters was analyzed. Both Wnt-11 and squamous cell carcinoma antigen were more commonly expressed in cervical cancer than in cervical intraepithelial neoplasia and in normal cervical tissue (respectively; P < 0.05); further, Wnt-11 and squamous cell carcinoma antigen expression in cervical cancer were positively correlated (r = 0.271, P < 0.05). In comparing the expression with clinicopathological parameters of tumor samples, Wnt-11 and squamous cell carcinoma antigen were both associated with FIGO stage, lymph node metastasis, and tumor size (P < 0.05), but not with patient age, pathological type, or differentiation. Increased Wnt-11 protein levels in cervical carcinoma samples were associated with a poor outcome in univariate and multivariate analysis. Wnt-11 and squamous cell carcinoma antigen are related to the malignancy degree and metastasis of cervical cancer, and thus may play a coordinating role in the occurrence and progression of cervical cancer. The study indicated that Wnt-11 may be a useful prognostic indicator for cervical carcinoma.