New mechanisms and biomarkers of lymph node metastasis in cervical cancer: reflections from plasma proteomics.

OBJECTIVE: Lymph node metastasis (LNM) and lymphatic vasculature space infiltration (LVSI) in cervical cancer patients indicate a poor prognosis, but satisfactory methods for diagnosing these phenotypes are lacking. This study aimed to find new effective plasma biomarkers of LNM and LVSI as well as possible mechanisms underlying LNM and LVSI through data-independent acquisition (DIA) proteome sequencing. METHODS: A total of 20 cervical cancer plasma samples, including 7 LNM-/LVSI-(NC), 4 LNM-/LVSI + (LVSI) and 9 LNM + /LVSI + (LNM) samples from a cohort, were subjected to DIA to identify differentially expressed proteins (DEPs) for LVSI and LNM. Subsequently, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed for DEP functional annotation. Protein-protein interaction (PPI) and weighted gene coexpression network analysis (WGCNA) were used to detect new effective plasma biomarkers and possible mechanisms. RESULTS: A total of 79 DEPs were identified in the cohort. GO and KEGG analyses showed that DEPs were mainly enriched in the complement and coagulation pathway, lipid and atherosclerosis pathway, HIF-1 signal transduction pathway and phagosome and autophagy. WGCNA showed that the enrichment of the green module differed greatly between groups. Six interesting core DEPs (SPARC, HPX, VCAM1, TFRC, ERN1 and APMAP) were confirmed to be potential plasma diagnostic markers for LVSI and LNM in cervical cancer patients. CONCLUSION: Proteomic signatures developed in this study reflected the potential plasma diagnostic markers and new possible pathogenesis mechanisms in the LVSI and LNM of cervical cancer.

ECT2 promotes malignant phenotypes through the activation of the AKT/mTOR pathway and cisplatin resistance in cervical cancer.

Epithelial cell transforming sequence 2 (ECT2) is expressed at high levels in various malignancies and contributes to malignant phenotypes in cancers. However, ECT2 is still not fully understood regarding its function and carcinogenic mechanism in cervical cancer. This research indicated that ECT2 expression was elevated in cervical cancer based on bioinformatics analysis and clinical specimens. Experiments in vitro and in vivo confirmed that ECT2 knockdown could suppress the proliferation and metastasis of cervical carcinoma cells. In addition, we found that silencing ECT2 could enhance the sensitivity to cisplatin and promote cell apoptosis. Mechanistically, we observed that ECT2 knockdown could inhibit the AKT/mTOR pathway and activate apoptosis, while ECT2 overexpression induced the opposite effect. The relationship between ECT2 and AKT was further confirmed by immunoprecipitation and rescue experiments. We found that the ECT2 and AKT could interact to form a complex, and knockdown AKT could offset all of the effects induced by ECT2. Our study emphasized the key point of ECT2 in the reversal of cisplatin resistance, and ECT2 could become a potential therapeutic target in cervical cancer.

The efficacy of medical treatment for adenomyosis after adenomyomectomy.

AIMS: To compare the efficacy of gonadotropin-releasing hormone agonist (GnRH-a) and GnRH-a + levonorgestrel-releasing intrauterine system (LNG-IUS) after adenomyomectomy for improved adenomyosis-associated symptoms. METHODS: Overall, 193 patients with adenomyosis included in this study were categorized into three groups: adenomyomectomy (n = 57, group 1), adenomyomectomy + GnRH-a (n = 83, group 2) and adenomyomectomy + GnRH-a + LNG-IUS (n = 53, group 3). Visual Analog Scale (VAS) scores and uterine volumes were determined to evaluate the severity of adenomyosis. Dysmenorrhea improvement and uterine volume were the main outcomes. RESULTS: The VAS scores of all patients reduced from 7.3 (6.0, 8.5) to 0 (0, 0.6) at the 6 months after surgery, which were significantly higher in group 1 compared to other groups (P < 0.05). In groups 1, 2 and 3, there were 14, 7 and 4 patients, respectively, who suffered dysmenorrhea recurrence. The mean recurrent-free-survival (RFS) was 51.6 ± 2.4, 58.0 ± 1.2 and 58.3 ± 1.0 months, respectively, which was significantly shorter in group 1 (P < 0.05). The dysmenorrhea recurrences were 26.3%, 6.1%, 5.9% in groups 1, 2 and 3, respectively, at the 36 months, which was significantly higher in group 1 (P < 0.01). Significantly decreased uterine volumes were observed in all patients from 222.2 (147.6, 350.4) to 77.0 (65.9, 94.1) mL (P < 0.05) at the 6 month after surgery. CONCLUSION: Treatment GnRH-a and LNG-IUS after surgery could significantly reduce the recurrence and prolong the RFS. It seemed that the use of LNG-IUS was beneficial for a lower recurrence in long-term follow-up.

Macrophages alternatively activated by endometriosis-exosomes contribute to the development of lesions in mice.

STUDY QUESTION: Do exosomes play a role in the pathogenesis of endometriosis in a murine model? SUMMARY ANSWER: Exosomes from endometriosis (EMS) can alternatively activate macrophages and thus contribute to the development of lesions in mice. WHAT IS KNOWN ALREADY: The pathogenesis of endometriosis, an inflammatory disease, possibly involves peritoneal macrophages. Exosomes are recognized as a new communicator among cells and a key modulator in several inflammatory diseases. STUDY DESIGN, SIZE, DURATION: We performed in vitr and in vivo experiments to demonstrate the role of exosomes in modulating macrophages. RAW264.7 cells (macrophages) were used to examine the effects of exosomes on macrophages in vitro. An experiment was also conducted in vivo, as follows. Fifty C57BL/6 female mice were randomly allocated to five control and five experimental groups (n = 5/group). The experimental group was injected i.p. with EMS-exosomes derived from eutopic stromal cells, starting on Day-7 then every day for 1 week. The control group received CON-exosomes from mice without endometriosis. Peritoneal macrophages were assessed over the next 6 days. On Day 0, all mice were injected i.p. with endometrium to establish the endometriosis model. On Day 14, all mice were sacrificed, ectopic lesions were counted and measured. PARTICIPANTS/MATERIALS, SETTING, METHODS: Exosomes were isolated from endometrial stromal cells (ESCs) by ultracentrifugation and characterized through transmission electron microscopy, nanoparticle tracking analysis and western blot. After treatment with exosomes, the polarization and phagocytic ability of the macrophages were detected by flow cytometry analysis, immunofluorescent staining and RT-PCR. C57BL/6 mice were utilized to establish an endometriosis model by i.p. injection of endometrial segments. MAIN RESULTS AND THE ROLE OF CHANCE: After treatment with EMS-exosomes, the macrophages were polarized into an M2-like phenotype and their phagocytic ability decreased (P < 0.05 versus treatment with CON-exosomes). The total weight and volume of the lesions in mice treated with EMS-exosomes significantly increased compared with those in mice treated with CON-exosomes (P < 0.05). The infiltration of M2-like macrophages was enhanced in the EMS-exosome group (P < 0.001 versus treatment with CON-exosomes). LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: Detection of endometriosis following exosome treatment was only performed in a murine endometriosis model. Clinical data and additional mechanism studies must be conducted to understand the role of exosomes in the pathogenesis of endometriosis. WIDER IMPLICATIONS OF THE FINDINGS: This study emphasizes the importance of EMS-exosomes in the pathogenesis of endometriosis. Further investigations on the exosome signaling pathways may contribute to the development of effective treatments for endometriosis. STUDY FUNDING/COMPETING INTEREST(S): This research was supported by grants (Nos. 81571417 and 81771552) from the National Science Foundation of China. The authors report no conflict of interest.