Yan Gao

Controversy on Positive Peritoneal Cytology of Endometrial Carcinoma

Endometrial carcinoma (EC) is one of the most common gynecological malignancies. Its incidence rate has been increasing year by year. The prognostic factors and treatment strategies of EC have aroused wide concern. The effects of peritoneal cytology on the prognosis and treatment of EC remain controversial. Some factors, such as differentiation degree, muscle invasion, and tumor size, are related to positive peritoneal cytology. Hysteroscopy is commonly used in the diagnosis and treatment of endometrial cancer, but hysteroscopic surgery may cause the tumor to spread into the abdominal cavity, resulting in positive peritoneal cytology. In this review, we discuss the factors related to positive peritoneal cytology and the influence of positive peritoneal cytology on the prognosis of endometrial cancer. Suspicious positive peritoneal cytology may be an independent risk factor for endometrial cancer. The positive rate of peritoneal tumor cells in type II endometrial cancer is higher than other cells and is an independent risk factor for type II endometrial cancer. We also discuss the effects of peritoneal cytology on treatment decisions. Aggressive treatments seem to be more beneficial for patients with positive ascites cytology, but there is a lack of large-scale prospective clinical studies on their effectiveness and safety. The application of peritoneal cytology for endometrial cancer has been decreased in recent years. We believe that peritoneal cytology is necessary for this type of cancer. However, more studies on peritoneal cytology in endometrial cancer should be carried out.

Hypoxia Triggers ALYREF ‐Mediated m 5 C Methylation of KIF20A to Activate KIF20A / BUB1 for Generating Ferroptosis Resistance in Cervical Cancer Cells

ABSTRACT Ferroptosis resistance is a key player in cervical cancer (CC) development. Hypoxia is a negative factor affecting CC treatment by inducing ferroptosis resistance. Our study aimed to investigate the detailed mechanisms of hypoxia‐induced ferroptosis resistance in CC cells. The mRNA and protein levels were assessed using RT‐qPCR and western blot. Immunofluorescence staining was used to analyze the co‐localization of Budding uninhibited by benzimidazole 1 (BUB1) and Kinesin family member 20A (KIF20A) in CC cells. Fe 2+ , MDA, and GSH levels were measured by commercial kits. Cell viability was determined by CCK‐8. The molecular interactions were analyzed by RIP or Co‐IP. MeRIP was adopted to measure the m 5 C level of KIF20A. We found that hypoxia facilitated ferroptosis resistance in CC cells. Hypoxia led to the upregulation of KIF20A, and KIF20A knockdown weakened hypoxia‐mediated ferroptosis resistance in CC cells. Mechanistically, Aly/REF export factor (ALYREF) stabilized KIF20A mRNA stability via m 5 C modification. In addition, KIF20A upregulated BUB1 in CC cells by directly interacting with BUB1. Rescue experiments indicated that BUB1 overexpression partially reversed the inhibitory effect of KIF20A knockdown on hypoxia‐mediated ferroptosis resistance in CC cells. In conclusion, hypoxia triggers ALYREF‐mediated m 5 C methylation of KIF20A to activate the KIF20A/BUB1 axis, thereby inducing ferroptosis resistance in CC cells.

Single‐cell transcriptomics identify TNFRSF1B as a novel T‐cell exhaustion marker for ovarian cancer

AbstractBackground:Ovarian cancer (OC) patients routinely show poor immunotherapeutic response due to the complex tumour microenvironment (TME). It is urgent to explore new immunotherapeutic markers.Methods:Through the single‐cell RNA sequencing (scRNA‐seq) analyses on high‐grade serous OC (HGSOC), moderate severity borderline tumour and matched normal ovary, we identified a novel exhausted T cells subpopulation that related to poor prognosis in OC. Histological staining, multiple immunofluorescences, and flow cytometry were applied to validate some results from scRNA‐seq. Furthermore, a tumour‐bearing mice model was constructed to investigate the effects of TNFRSF1B treatment on tumour growth in vivo.Results:Highly immunosuppressive TME in HGSOC is displayed compared to moderate severity borderline tumour and matched normal ovary. Subsequently, a novel exhausted subpopulation of CD8+TNFRSF1B+ T cells is identified, which is associated with poor survival. In vitro experiments demonstrate that TNFRSF1B is specifically upregulated on activated CD8+ T cells and suppressed interferon‐γ secretion. The expression of TNFRSF1B on CD8+T cells is closely related to OC clinical malignancy and is a marker of poor prognosis through 140 OC patients’ verification. In addition, the blockade of TNFRSF1B inhibits tumour growth via profoundly remodeling the immune microenvironment in the OC mouse model.Conclusions:Our transcriptomic results analyzed by scRNA‐seq delineate a high‐resolution snapshot of the entire tumour ecosystem of OC TME. The major applications of our findings were an exhausted subpopulation of CD8+TNFRSF1B+ T cells for predicting OC patient prognosis and the potential therapeutic value of TNFRSF1B. These findings demonstrated the clinical value of TNFRSF1B as a potential immunotherapy target and extended our understanding of factors contributing to immunotherapy failure in OC.

The Overexpression of Keratin 23 Promotes Migration of Ovarian Cancer via Epithelial‐Mesenchymal Transition

Background. Keratin 23 (KRT23) is a new member of the KRT gene family and known to be involved in the development and migration of various types of tumors. However, the role of KRT23 in ovarian cancer (OC) remains unclear. This study is aimed at investigating the function of KRT23 in OC. Methods. The expression of KRT23 in normal ovarian and OC tissues was determined using the Oncomine database and immunohistochemical staining. Reverse transcription quantitative polymerase chain reaction assay was used to analyze the expression of KRT23 in normal ovarian epithelial cell lines and OC cell lines. Small interfering RNA (siRNA), wound healing assay, and transwell assay were conducted to detect the effects of KRT23 on OC cell migration and invasion. Further mechanistic studies were verified by the Gene Expression Profiling Interactive Analysis platform, Western blotting, and immunofluorescence staining. Results. KRT23 was highly expressed in OC tissues and cell lines. High KRT23 expression could regulate OC cell migration and invasion, and the reduction of KRT23 by siRNA inhibited the migration and invasion of OC cells in vitro. Furthermore, KRT23 mediated epithelial‐mesenchymal transition (EMT) by regulating p‐Smad2/3 levels in the TGF‐β/Smad signaling pathway. Conclusions. These results demonstrate that KRT23 plays an important role in OC migration via EMT by regulating the TGF‐β/Smad signaling pathway.

Single-cell RNA-seq highlights a specific carcinoembryonic cluster in ovarian cancer

AbstractExpounding the heterogeneity for ovarian cancer (OC) with the cognition in developmental biology might be helpful to search for robust prognostic markers and effective treatments. In the present study, we employed single-cell RNA-seq with ovarian cancers, normal ovary, and embryo tissue to explore their heterogeneity. Then the differentiation process of clusters was explored; the pivotal cluster and markers were identified. Furthermore, the consensus clustering algorithm was used to explore the different clinical phenotypes in OC. At last, a prognostic model was construct and used to assess the prognosis for OCs. As a result, eight diverse clusters were identified, and the similarity existed in some clusters between embryo and tumours based on their gene expression. Meaningfully, a subtype of malignant epithelial cluster, PEG10+ EME, was associated with poor survival and was an intermediate stage of embryo to tumour. PEG10 was a CSC marker and might influence CSC self-renewal and promote cisplatin resistance via NOTCH pathway. Utilising specific gene profiles of PEG10+ EME based on public data sets, four phenotypes with different survival and clinical response to anti-PD-1/PD-L1 immunotherapy were identified. These insights allowed for the investigation of single-cell transcriptome of OCs and embryo, which advanced our current understanding of OC pathogenesis and resulted in promising therapeutic strategies.