Xin yixue (Dec 2022)
Exosomal miR-21-5p derived from metastatic cervical cancer cells promotes cervical cancer invasion and metastasis
Abstract
Objective To explore the influence of paracrine function of recurring and metastatic cervical cancer tissues on the spread of cancer cells. Methods Surgical specimens （cancerous and para-cancerous tissues） were obtained from 22 patients with metastatic cervical squamous cancer. Serum specimens were collected from 22 patients with metastatic cervical squamous cancer and 22 healthy volunteers， respectively. The expression levels of miR-21-5p in the cancerous and para-cancerous tissues were determined by quantitative real-time PCR （RT-qPCR）. SiHa cell were transferred with miR-21-5p mimics/inhibitors， respectively. The differences between two groups were analyzed by RT-qPCR， colony formation test， Transwell migration test， MTT， western blotting and dual luciferase reporter assay. Serum exosomes were extracted by high-speed centrifugation. The exosome characterization was performed by nanoparticle tracking analysis （NTA）， western blotting and transmission electron microscopy （TEM）. The differences in the expression levels of miR-21-5p in serum exosomes between cervical squamous cancer patients and healthy volunteers were assessed by RT-qPCR. The diagnostic efficiency of miR-21-5p in serum exosomes was evaluated by the receiver operating characteristic （ROC） curve. Results The expression level of miR-21-5p was up-regulated in cervical cancerous tissues compared with para-cancerous tissues. And over-expressed miR-21-5p could significantly promote SiHa cell colony formation， cell proliferation and migration （all P <0.01）. Dual luciferase reporter assay showed that miR-21-5p could target SPYR2 and regulate the expression level of SPYR2. In SiHa cells transfected with miR-21-5p mimics， the expression levels of p-MSK， p-90RSK， p-C-RAF， p-MEK and p-MAPK proteins were significantly up-regulated （all P < 0.05）. The expression level of miR-21-5p was significantly up-regulated in serum exosomes of patients with cervical squamous cancer （P < 0.05）. The area under the ROC curve （AUC） of serum exosomes for the assessment of cervical cancer metastasis was 0.9375. Conclusions The transfer of miR-21-5p to exosomes in the tissues of patients with metastatic cervical squamous cancer promotes the spread of cancer cells. The mechanism may be related to the targeting of the SPRY2/ERK/MAPK signaling pathway.
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