Xiaojing Chen

Knockdown of TRIM47 Overcomes Paclitaxel Resistance in Ovarian Cancer by Suppressing the TGF‐β Pathway via PPM1A

ABSTRACTObjectiveWe investigated the role of tripartite motif 47 (TRIM47) in paclitaxel resistance in ovarian cancer, focusing on its regulation of protein phosphatase magnesium‐dependent 1A (PPM1A), transforming growth factor‐β (TGF‐β) pathway activation, and methyltransferase‐like 3 (METTL3)‐mediated N6‐methyladenosine (m6A) modification.MethodsBioinformatics analysis using Gene Expression Profiling Interactive Analysis (GEPIA), Kaplan–Meier plotter (KM plot), Linkedomics, and sequence‐based RNA adenosine methylation site predictor (SRAMP) databases identified TRIM47 and PPM1A expression patterns, prognostic significance, co‐expression networks, and m6A modification sites. Paclitaxel‐resistant ovarian cancer cell lines were generated. Quantitative reverse transcriptase polymerase chain reaction (qRT‐PCR) and western blot analyzed gene and protein expression. Co‐immunoprecipitation (Co‐IP) and GST pull‐down assays assessed TRIM47‐PPM1A interaction, while cycloheximide (CHX) chase, and IP assays examined PPM1A stability and ubiquitination. RNA immunoprecipitation (RIP) and dual‐luciferase assays determined METTL3’s effect on TRIM47 m6A modification. Functional assays (3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT), colony formation, and flow cytometry) evaluated proliferation, apoptosis, and drug response. An in vivo xenograft model confirmed TRIM47’s role in chemoresistance.ResultsBioinformatics analysis showed that TRIM47 was overexpressed in ovarian cancer and negatively correlated with the expression of PPM1A. Kaplan–Meier analysis showed that high TRIM47 and low PPM1A expression were correlated with poor prognosis. TRIM47 was upregulated in paclitaxel‐resistant ovarian cancer cells. The knockdown of TRIM47 restored drug sensitivity, inhibited cell proliferation, and induced cell apoptosis. Mechanistically, TRIM47 functioned as an E3 ubiquitin ligase, targeting PPM1A for degradation, leading to sustained TGF‐β signaling and enhanced chemoresistance. CHX chase assays demonstrated reduced PPM1A stability in the presence of TRIM47, while IP‐WB confirmed increased PPM1A ubiquitination. METTL3‐mediated m6A modification enhanced TRIM47 mRNA stability, further promoting its oncogenic role. In vivo, TRIM47 knockdown reduced tumor growth and improved paclitaxel efficacy, reinforcing its role in resistance.ConclusionTRIM47 promoted paclitaxel resistance in ovarian cancer by inducing PPM1A degradation and activating the TGF‐β pathway.

Effects of long-acting versus short-acting granulocyte colony stimulating factor after radiotherapy in gynecologic malignancies: a prospective observational cohort study

Little is known about the role of the protective effects of granulocyte colony-stimulating factor (G-CSF) in patients after radiotherapy. The aim of the present study was to explore the prophylactic effects of long-acting granulocyte colony-stimulating factor (G-CSF) on febrile neutropenia (FN) and myelosuppression in chemotherapy patients with gynecologic malignancies after pelvic radiotherapy. Patients voluntarily participated in a study group (long-acting G-CSF for all chemotherapy cycles) and a control group (short-acting G-CSF) after they were educated about G-CSF utilization. The incidences of FN and myelosuppression, as well as adverse events, were compared between the two groups. A regression model was used to determine the risk factors for FN and myelosuppression. From January 6, 2019, to August 22, 2019, 61 patients were included in the final analysis, with 286 chemotherapy cycles. There were 14 (23.0%) and 57 (77.0%) patients in the study and control groups, respectively. The study group had significantly fewer complete blood count tests, fewer outpatient clinic visits, fewer short-acting G-CSF doses, and lower incidences of FN and myelosuppression per chemotherapy cycle. According to the binary regression model, the use of long-acting G-CSF was the only factor associated with a decreased incidence of myelosuppression but not FN. The major adverse event related to G-CSF was mild bone pain. In conclusion, long-acting G-CSF may effectively reduce the incidence of FN and myelosuppression with mild adverse effects during chemotherapy after radiotherapy.Trial registrationRegistered at https://www.clinicaltrials.gov/  on January 4, 2019 (NCT03793205).

Fragmentomics features of ovarian cancer

AbstractOvarian cancer (OC) is a major cause of cancer mortality in women worldwide. Due to the occult onset of OC, its nonspecific clinical symptoms in the early phase, and a lack of effective early diagnostic tools, most OC patients are diagnosed at an advanced stage. In this study, shallow whole‐genome sequencing was utilized to characterize fragmentomics features of circulating tumor DNA (ctDNA) in OC patients. By applying a machine learning model, multiclass fragmentomics data achieved a mean area under the curve (AUC) of 0.97 (95% CI 0.962–0.976) for diagnosing OC. OC scores derived from this model strongly correlated with the disease stage. Further comparative analysis of OC scores illustrated that the fragmentomics‐based technology provided additional clinical benefits over the traditional serum biomarkers cancer antigen 125 (CA125) and the Risk of Ovarian Malignancy Algorithm (ROMA) index. In conclusion, fragmentomics features in ctDNA are potential biomarkers for the accurate diagnosis of OC.