Transforming growth factor (TGF)-β1 is a key cytokine affecting the pathogenesis and progression of cervical cancer. Tumor-derived 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 TGF-β1 affects tumor biological functions through exosomes released by cervical cancer cells. Deep RNA sequencing found that TGF-β1 stimulated cervical cancer cells to secrete more miR-663b-containing exosomes, which could be transferred into new target cells to promote metastasis. Further studies have shown that miR-663b directly targets the 3′-untranslated regions (3′-UTR) of mannoside acetylglucosaminyltransferase 3 (MGAT3) and is involved in the epithelial-mesenchymal transition (EMT) process. Remarkably, the overexpression of MGAT3 suppressed cervical cancer cell metastasis promoted by exosomal miR-663b, causing increased expression of epithelial differentiation marker E-cadherin and decreased expression of mesenchymal markers N-cadherin and β-catenin. Throughout our study, online bioinformation tools and dual luciferase reporter assay were applied to identify MGAT3 as a novel direct target of miR-663b. Exosome PKH67-labeling experiment verified that exosomal miR-663b 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 miR-663b and MGAT3 and the regulation of the EMT pathway caused by MGAT3 were detected by quantitative real-time transcription-polymerase chain reaction (qPCR) and western blot analysis. These results, thus, provide evidence that cancer cell-derived exosomal miR-663b is endocytosed by cervical cancer cells adjacent or distant after TGF-β1 exposure and inhibits the expression of MGAT3, thereby accelerating the EMT process and ultimately promoting local and distant metastasis.

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.

Objective: This study aimed to identify reliable risk factors for residual/recurrent cervical intraepithelial lesions in patients with negative margins after cold-knife conization. Methods: A total of 2352 women with HSILs (high-grade squamous intraepithelial lesions) with negative margins who underwent cold-knife conization between January 2014 and December 2020 were included; in total, 1411 women were assigned to the development cohort, and 941 women were assigned to the validation cohort. Multivariate logistic regression was used to build four predictive models based on the different combinations of follow-up data (Model A: preoperative factors; Model B: first-follow-up data; Model C: second-follow-up data; Model D: data from both follow-ups). The accuracy, sensitivity, specificity, false-positive rate (FPR), false-negative rate (FNR), and area under the receiver operating characteristic curve (AUC) were evaluated on the validation cohort. The predictive power of risk factors was further validated using six machine learning algorithms. Results: Model D demonstrated the highest AUC of 0.91 (95% CI, 0.87 to 0.96) in the validation cohort, whereas Models A, B, and C achieved AUCs of 0.69 (95% CI, 0.59 to 0.78), 0.88 (95% CI, 0.80 to 0.95), and 0.89 (95% CI, 0.81 to 0.97) respectively. The six machine learning methods achieved consistent results. Kaplan-Meier (KM) survival curves demonstrated that our models could effectively stratify patients with all models (p < 0.05 for all models). Conclusion: Our model, which is based on preoperative and follow-up factors, can serve as a complementary screening procedure for the early detection or prediction of recurrence after cold-knife conization in HSIL patients.