Micro (mi)RNAs are crucial participants in the progression of cervical cancer (CC). Growing evidence indicates that miRNA (miR)-34c-5p is a pivotal tumor suppressor in numerous types of cancer and its functions in CC require further investigating. The present study demonstrated that there was a decreased level of miR-34c-5p in CC-associated cell lines compared with healthy control samples. It also demonstrated that miR-34c-5p targeted Notch1 and suppressed CC progression. Dual-Luciferase reporter assays verified the targeted relationship of miR-34c-5p and Notch1. The expression of Notch1 in HeLa cells was markedly reduced following miR-34c-5p overexpression and the proliferation, migration and invasion of HeLa cells were reduced although apoptosis was accelerated. However, overexpression of miR-34c-5p was reversed following the addition of Notch1, which supported the finding of the targeted relationship between miR-34c-5p and Notch1. Flow cytometry demonstrated that miR-34c-5p inhibited the proliferation of HeLa cells while accelerating apoptosis. The present study concluded that miR-34c-5p was a tumor suppressor in CC and may be a novel measure for the future treatment of CC.

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This study tried to evaluate whether 8% polyethylene glycol (PEG) 6000 precipitation combined with differential ultracentrifugation (PPDU) was an efficient and practical method for the enrichment and purification of extracellular vesicles (EVs) derived from the culture supernatant of human ovarian cancer cell line A2780 and from body fluids of patients with high-grade serous carcinoma (HGSC). Methods: PPDU was used to enrich and purify the EVs derived from body fluids of patients with HSGC and cell culture supernatant of subclones of human ovarian cancer cell line A2780 with high/low invasive capacity (named as A-H/A-L, respectively). Transmission electron microscope (TEM) and nanoparticle tracking analysis (NTA) were used to identificate the EVs size and distribution. Western blots (WB) were used to detect the expression of CD9, CD63, Alix and Calnexin. The high-purity EVs derived from the cell culture supernatant of A-H/A-L were detected by the protein profile. Expression of integrins (ITGs) αV, β1 and β3 in the EVs derived from body fluids of HGSC patients was also evaluated. Results: The diameter of EVs was about 30-260 nm observed under the TEM. Under the NTA identification, the peak size of EVs was ranged from 70 to 159nm. EVs derived from different specimens did not significantly differ in mean size and peak size. Presence of CD9, CD63 and Alix and absence of Calnexin were confirmed in the EVs. The protein concentrations of EVs' sample extracted from A-H/A-L cell culture supernatant were 0.36µg/µL and 0.20µg/µL, respectively. The total amount of protein obtained from 300ul EVs was 108.02ug and 61.44ug, respectively. Totally, 2397 peptides and 952 proteins were identified by isobaric tags for relative and absolute quantitation (ITRAQ). The expression of ITGαV, β1, and β3 in the EVs from plasma and ascites of HGSC patients was significantly higher than the control group (plasma: all P<0.0001; ascites: P=0.036, 0.001 and 0.004, respectively). The expression level of ITGαV and β1 in EVs of HGSC's ascites was significantly higher than that in plasma (P= 0.004, 0.001, respectively). The expression of ITGβ3 was also slightly elevated in EVs-derived HGSC patients' ascites (P=0.492). Conclusion: PPDU was an efficient and practical method to enrich EVs from body fluids and cell culture supernatant. The characteristic expression of ITGαV, β1 and β3 in ascites and plasma EVs of patients with HGSC provided useful information on the development of EVs in HGSC.

Background Intra-tumoral heterogeneity (ITH) has resulted in treatment failure of ovarian cancer(OC). Exosomes and microRNA plays a crucial role in the progression of OC. Therefore, we aimed to explore the effect of exosomes and microRNA421 (miR-421), mediated by exosomes, on the ITH by activating the PI3K/AKT pathway and the diagnosis of OC. Method Exosomes derived from AHC/ALC cells (AHE/ALE) were extracted by differential centrifugation. CCK8, 5-ethyl-2'-deoxyridine(EdU), Transwell, Colony formation and Wound healing assays were performed to explore the proliferation, invasion, and migration abilities. Western blot (WB) assay was used to detect the changes in the epithelial-mesenchymal transition (EMT) and PI3K/AKT pathway. Immunofluorescence assay was used to detect changes in EMT. qRT-PCR was used to detect microRNA levels in serum exosomes from high grade serous ovarian cancer (HGSOC) and benign patients. We also measured the levels of CA125 in serum exosomes. Result AHE and miR-421, mediated by exosomes, significantly increased the malignancy of ALC cells by activating the PI3K/AKT pathway. the expression of miR-421 was significantly increased in the serum exosomes derived from HGSOC patients. Receiver operating characteristic (ROC) curve analysis showed that the combination of miR-421, and serum CA125 can significantly improve the specificity of serum CA125 in the diagnosis of HGSOC. Conclusion MiR-421, mediated by exosomes, could induce the transformation of high-invasive cell subpopulations from low-invasive cell subpopulations of OC cells by activating the PI3K/AKT pathway. MiR-421 could serve as a potentially effective therapeutic target and a novel tumor marker for early diagnosis of OC.