The comparison of pure uterine serous carcinoma and mixed tumor with serous component: a single-institution review of 91 cases.

BACKGROUND: Pure uterine serous carcinoma (p-USC) and mixed tumors with serous component (m-USC) are aggressive subtypes of endometrial cancer associated with high mortality rates. This retrospective study aimed to compare clinicopathologic features and outcomes of p-USC and m-USC in a single center. METHODS: This study retrospectively reviewed patients diagnosed with USC at Peking University People's Hospital between 2008 and 2022. T-tests and chi-square tests were used to compare clinicopathological characteristics between p-USC and m-USC. Kaplan-Meier survival curve and Cox regression analysis were used to analyze the impact of clinical and pathological variables on OS and PFS. RESULTS: Among the 91 patients who underwent surgery, 65.9% (n = 60) were p-USC, and 34.1% (n = 31) were m-USC. Patients with p-USC had earlier menopause (P = 0.0217), a lower rate of progesterone receptor(PR) expression (P < 0.001), and were more likely to have positive peritoneal cytology (P = 0.0464). After a median follow-up time of 40 months, 28 (46.7%) p-USC and 9 (29%) m-USC patients had progression disease, 18 (30%) and 8 (25.8%) patients died of their disease. 5-year PFSR were 51.2% and 75.3%, respectively, and 5-year OS rates were 66% and 67.4%. Kaplan-Meier survival analysis showed that p-USC was more likely to relapse than m-USC (P = 0.034), but there was no significant difference in OS. Cox regression analysis showed that lymph node metastasis and surgical approach were risk factors for OS, and myoinvasion depth ≥ 1/2 was an independent risk factor for PFS. CONCLUSIONS: p-USC was more likely to relapse than m-USC, but there was no significant difference in OS between the two subtypes.

Cervical cancer-derived exosomal miR-663b promotes angiogenesis by inhibiting vinculin expression in vascular endothelial cells.

BACKGROUND: Angiogenesis provides essential nutrients and oxygen for tumor growth and has become the main mechanism of tumor invasion and metastasis. Exosomes are nanoscale membrane vesicles containing proteins, lipids, mRNA and microRNA (miRNA), which mediate intercellular communication and play an important role in tumor progression. Accumulated evidence indicates that tumor-derived exosomal miRNAs participate in the tumor microenvironment and promote angiogenesis. METHODS: Bioinformatic target prediction and dual luciferase reporter assays were performed to identify the binding site between miR-663b and the 3'-UTR of vinculin (VCL). VCL overexpression lentivirus and miR-663b overexpression/inhibition lentivirus were used to create a VCL overexpression model and miR-663b overexpression/inhibition model in-vitro. Immunohistochemistry (IHC) assays and western blot assays were used to detect protein expression. Exosome-cell cocultures, wound healing assays, tube formation assays and transwell assays were used to measure the migration and tube formation ability of vascular endothelial cells [human umbilical vein endothelial cells (HUVECs)]. siRNA targeted VCL was used to knockdown VCL. RESULTS: In the present study, we found that miR-663b was elevated in cervical cancer tissue and exosomes. miR-663b could bind the 3'-UTR of VCL and inhibit its expression. VCL is downregulated in cervical cancer, and decreased VCL has a negative correlation with a high level of miR-663b. Further studies demonstrated that exosomes secreted by cervical cancer cells can deliver miR-663b to HUVECs and inhibit the expression of VCL, thereby promoting angiogenesis and tumor growth. CONCLUSIONS: miR-663b derived from cancer cell exosomes acts as a driving factor for angiogenesis and a potential target of antiangiogenic therapy in cervical cancer. Our findings illustrated a new signaling pathway, including exosomes, miRNAs and target genes, which provides potential targets for antiangiogenic therapy.

CD155 Promotes the Progression of Cervical Cancer Cells Through AKT/mTOR and NF-κB Pathways.

Expression of the immunoglobulin superfamily member CD155 was increased in a variety of human malignancies, but the role of CD155 in tumorigenesis and tumor development in cervical cancer has not been elucidated. In this study, immunohistochemistry and enzyme-linked immunosorbent assay analyses showed that CD155 expression gradually increases with the degree of cervical lesions. In vitro and in vivo, reducing the expression of CD155 inhibited cell proliferation, cell viability and tumor formation and arrested the cell cycle in G0/G1 phase. Antibody array-based profiling of protein phosphorylation revealed that CD155 knockdown can inhibited the AKT/mTOR/NF-κB pathway and activated autophagy and apoptosis; the opposite effects were observed upon CD155 has overexpression. We proved that there is an interaction between CD155 and AKT by immunoprecipitation. We further confirmed the mechanism between CD155 and AKT/mTOR/NF-κB through rescue experiments. AKT knockdown reversed the anti-apoptotic effects and activation of the AKT/mTOR/NF-κB pathway induced by CD155 overexpression. Our research demonstrated that CD155 can interact with AKT to form a complex, activates the AKT/mTOR/NF-κB pathway and inhibit autophagy and apoptosis. Thus, CD155 is a potential screening and therapeutic biomarker for cervical cancer.

Management of high-grade squamous intraepithelial lesion patients with positive margin after LEEP conization: A retrospective study.

ABSTRACT: To explore the optimal way to manage patients with high-grade squamous intraepithelial lesion (HSIL) and positive margin by identifying the risk factors for its recurrence and residue.A retrospective study was conducted on 267 cases of a pathologically confirmed HSIL with positive margin following conization by loop electrosurgical excisional procedure (LEEP) between January 2010 and December 2015. One hundred two cases were selected for regular follow-up every 6 months, and 165 cases were selected for a second surgery (repeat cervical conization or hysterectomy) within 3 months of initial LEEP. We analyzed the association between recurrent or residual diseases and these factors: age, menopausal status, ThinPrep cytologic test (TCT) results, high-risk human papillomavirus (HR-HPV) infection, pathological grades of the margin, number of involved margins, and glandular involvement.The recurrence rate among 102 cases who underwent follow-up was 17.6% (18/102). The factors: atypical squamous cells of undetermined significance cannot exclude HSIL (ASC-H) or higher lesions in the pre-LEEP TCT (P = .038), persistent HR-HPV infection at the 6th month post-LEEP (P = .03), HSIL-positive margin (P = .003), and multifocal-involved margin (P = .002) were significantly associated with recurrent disease, while age, menopause, and pre-LEEP HR-HPV infection were not associated with recurrent disease (P > .05). The residual rate among 165 patients who underwent a second surgery was 45.5% (75/165), of which 15 cases were residual cervical cancer. The factors: menopause (P = .02), ≥ASC-H in pre-LEEP TCT (P = .04), pre-LEEP HR-HPV infection (P = .04), ≥HSIL-positive margin (P < .001), and multifocal-involved margin (P < .001) significantly increased the risk of residual disease. No correlation existed between residual disease and age or glandular involvement (P > .05).For patients with a positive margin after LEEP, regular follow-up or second surgery should be selected according to fertility requirement and pathological characteristics of the positive margin, as well as TCT and HR-HPV infection condition.

Exosomal miR­663b exposed to TGF­ß1 promotes cervical cancer metastasis and epithelial­mesenchymal transition by targeting MGAT3.

Transforming growth factor (TGF)­ß1 is a key cytokine affecting the pathogenesis and progression of cervical cancer. Tumor­derived exosomes contain microRNAs (miRNAs/miRs) that interact with cancer and stromal cells, thereby contributing to tissue remodeling in the tumor microenvironment (TME). The present study was designed to clarify how TGF­ß1 affects tumor biological functions through exosomes released by cervical cancer cells. Deep RNA sequencing found that TGF­ß1 stimulated cervical cancer cells to secrete more miR­663b­containing exosomes, which could be transferred into new target cells to promote metastasis. Further studies have shown that miR­663b directly targets the 3'-untranslated regions (3'­UTR) of mannoside acetylglucosaminyltransferase 3 (MGAT3) and is involved in the epithelial­mesenchymal transition (EMT) process. Remarkably, the overexpression of MGAT3 suppressed cervical cancer cell metastasis promoted by exosomal miR­663b, causing increased expression of epithelial differentiation marker E­cadherin and decreased expression of mesenchymal markers N­cadherin and ß­catenin. Throughout our study, online bioinformation tools and dual luciferase reporter assay were applied to identify MGAT3 as a novel direct target of miR­663b. Exosome PKH67­labeling experiment verified that exosomal miR­663b could be endocytosed by cervical cancer cells and subsequently influence its migration and invasion functions which were measured by wound healing and Transwell assays. The expression of miR­663b and MGAT3 and the regulation of the EMT pathway caused by MGAT3 were detected by quantitative real­time transcription­polymerase chain reaction (qPCR) and western blot analysis. These results, thus, provide evidence that cancer cell­derived exosomal miR­663b is endocytosed by cervical cancer cells adjacent or distant after TGF­ß1 exposure and inhibits the expression of MGAT3, thereby accelerating the EMT process and ultimately promoting local and distant metastasis.

CD155/TIGIT, a novel immune checkpoint in human cancers (Review).

CD155/T cell immunoreceptor with Ig and ITIM domains (TIGIT) is a novel type of immune checkpoint. CD155 is an adhesion molecule that is upregulated during tumor progression and promotes the proliferative and migratory abilities of tumor cells via various pathways. TIGIT, an inhibitory receptor, is mainly expressed on natural killer (NK), CD8+ T, CD4+ T and T regulatory (Treg) cells. CD155 transmits immune signals via interacting with the inhibitory checkpoint receptor TIGIT, thereby inhibiting the function of T and NK cells. Several preclinical studies have supported the use of TIGIT blockade as a monotherapy or combined with other immune checkpoint inhibitors for the treatment of advanced solid malignant tumors. The present review summarized the current knowledge on CD155/TIGIT and the lymphocyte­mediated inhibitory mechanism of CD155/TIGIT. An in­depth understanding of the role of CD155/TIGIT in tumors may aid to improve the application of immune checkpoint inhibitors in tumor therapy.

Role and mechanism of FLNa and UCP2 in the development of cervical cancer.

Recent studies have reported that filamin A (FLNa) and uncoupling protein 2 (UCP2) are highly expressed in various types of cancer, but little is currently known about their roles in cervical cancer (CC). In the present study, immunohistochemical staining of paraffin sections of cervical tissues was performed in order to compare the differential expression of FLNa, UCP2, p16 and Ki67 between CC and high­grade intraepithelial neoplasia (HSIL). UCP2 and FLNa were knocked down in CC cell lines to investigate the effects on cell proliferation, cell cycle arrest, apoptosis, migration and invasion. In addition, the present study investigated the expression of cell­associated proteins [extracellular signal­regulated kinase (ERK), phosphorylated (p) ERK, protein kinase B (AKT), p­AKT and B­cell lymphoma­2 (Bcl­2)] and the mRNA levels of cellular proteins such as Ras, matrix metalloproteinase (MMP)­2 and MMP­9. FLNa and UCP2 expression levels were significantly higher in CC tissues than in HSIL tissues, with no significant differential expression of p16 or Ki67. UCP2 expression was significantly different in patients with clinical stage II or higher or lymph node metastasis compared with in other patients with cervical cancer. FLNa or UCP2 knockdown slowed or decreased SiHa and HeLa cell proliferation, migration and invasion, with no significant change in apoptosis, and downregulated the protein levels of p­ERK1/2, and the mRNA levels of Ras, MMP­2 and MMP­9. UCP2 knockdown arrested the cell cycle at the G2 phase in SiHa and HeLa cells, while FLNa knockdown arrested the cell cycle at the G2 phase in HeLa cells. The results of the present study revealed that FLNa and UCP2 play roles in the development and progression of CC via the Ras/MAPK/ERK signalling pathway. FLNa and UCP2 are superior to p16 and Ki67 for early prediction of CC, indicating that FLNa and UCP2 may be used for the early diagnosis of CC. UCP2 may be used to predict the prognosis of CC.

Clinicopathologic significance and treatment of ASC-US in cervical cytology.

BACKGROUND: In China, cervical cancer is one of the most common gynecologic malignancies. Cervical cytology is an essential method for screening cervical cancer and cervical intraepithelial neoplasia (CIN), and the most common cytological abnormality result is atypical squamous cells of undetermined significance (ASC-US). Therefore, how to effectively deal with ASC-US cytology has become the focus of scholars. OBJECTIVE: We aim to analyze the final histopathologic results, clinicopathologic significance and current rationale of ASC-US cytology. METHODS: All patients with first ASC-US cytological reports who attended our gynecological outpatient clinic in Qilu Hospital of Shandong University during January 2010 to December 2015 were recruited to this study. The data were derived from clinical records and evaluated retrospectively. The results of age, High-Risk HPV (DNA) testing, colposcopy, and pathological outcomes were obtained. Directed biopsy was performed if there were any suspicious cervical lesions under colposcopy, while four quadrant biopsy and/or ECC were performed if no suspicious lesions were noted in colposcopies. RESULTS: A total of 1246 patients diagnosed with ASC-US were involved in the final statistical analysis. Mean age of patients was 41.6 years and the age range between 40-49 years represented 38.52% of all ASC-US women in this study. All patients were evaluated for HPV (DNA) and positive percent for High-Risk HPV was (67.1%). According to the final histopathologic outcomes after ASC-US cytology, 15.6% and 1.1% of patients had ≥ CIN2+ and invasive carcinoma respectively. Patients with invasive carcinoma were associated with HPV16+ and HPV18+. The detection rate of ≥ CIN2+ among the ASC-US/High-Risk HPV+ group was (53.9%) with a negative-predictive-value (NPV) of 100%. Our findings showed that the final pathologic results of ≥ CIN2+ were consistent with colposcopy with a coincidence rate of (77%), and colposcopical impression sensitivity and specificity for ≥ CIN2+ was (91.1% and 96.1%) respectively. CONCLUSION: women with ASC-US have a wide range of final pathologic results, and it can be the initial warning of high-grade CIN or cervical cancer. In China, HPV (DNA) testing triage is a useful shunting measure for ASC-US patients, and an immediate colposcopy is a consequential strategy for dealing with ASC-US cytology to increase the detection rate of high-grade cervical lesions or invasive cancer.

The role of low density lipoprotein receptor-related protein 11 as a tumor promoter in cervical cancer.

BACKGROUND: It is unclear whether low density lipoprotein receptor-related protein 11 (LRP11), a newly found lipoprotein receptor regulatory protein, has the carcinogenic effects in cervical cancer. METHODS: Bioinformatics analysis, immunohistochemical (IHC) staining and evaluation, cell proliferation assay, flow cytometry, transwell migration and invasion assays, Western blotting, growth of LRP11-silenced cells in athymic nude mice were performed in this research. RESULTS: We found that LRP11 expression was higher in high-grade squamous intraepithelial lesions (HSIL) and cervical cancer tissue than in normal cervix, and high expression of LRP11 was associated with differentiation degree (P=0.0266), indicating poor prognosis (P=0.0210). The silencing of LRP11 in SiHa and CaSki cell lines inhibited cell proliferation, reduced migration and invasion and suppressed cell growth in nude mice, which possibly related to cell cycle protein regulation of CDK 2/4, cyclin D1/E1, MMP-2/9, and VEGF. Furthermore, LRP11 showed substantial positive correlation with P16 in vivo and in vitro. CONCLUSION: LRP11 plays important roles in proliferation, migration and invasion, with the potential to be a useful prognostic marker and therapeutic target for patients with HSIL and cervical cancer.