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Clinical Research
miR-19 regulates PI3K/Akt signaling pathway by targeting PTEN to promote invasion and migration of endometrial cancer KLE cells
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Abstract:
Objective: To analyze the regulatory effects of miR-19 on PTEN and PI3K/Akt signaling pathway, and to reveal the effects of miR-19 and PTEN on invasion and migration of endometrial cancer KLE cells as well as its working mechanism. Methods: Tumor tissues and para-carcerous tissues of 74 patients with endometrial cancer were collected from the First Hospital of Hebei Medical University from May 2017 to August 2020. All patients did not receive radiotherapy or chemotherapy before surgery and were pathologically diagnosed as endometrial cancer. qPCR and WB were used to detect the effects of miR-19 mimics or inhibitors on PTEN expression in KLE cells. TargetScan and Dual-luciferase reporter gene assay were adopted to predict and verify whether PTEN was the target gene of miR-19. KLE cells were randomized into negative control (NC) group (transfected with miR-NC), miR-19 mimic group (transfected with miR-19 mimics) and miR-19+PTEN group (transfected with miR-19 mimics+PTEN overexpression vector).Transwell assay and Scratch assay were used to detect the effects of miR-19 and PTEN on KLE cell migration and invasion. WB was adopted to detect the expression of PI3K/Akt pathway-related proteins.Results: mRNA and protein expression of PTEN was obviously lower in endometrial cancer tissues compared with para-carcerous tissues (P<0.01). miR-19 could specifically bind to 3'-UTR of PTEN.Up-regulation of miR-19 could inhibit mRNA and protein expression of PTEN, and down-regulation of miR-19 showed an opposite result (P<0.01). Compared with miR-NC group, migratory cell count, invasive cell count, and scratch healing rate were significantly increased in miR-19 mimic group (all P<0.01). Compared with miR-19 mimic group, migratory cell count, invasive cell count and scratch healing rate were significantly decreased in miR-19+PTEN group (all P<0.01). WB analysis showed that, comparing with miR[1]NC group, PTEN protein was obviously decreased, while p-Akt 308 and p-Akt 473 protein was obviously elevated in miR-19 mimic group (P<0.01); however, Akt protein maintained unchanged. Additionally, the levels of p-Akt 308 and p-Akt 473 protein were obviously lower in miR-19+PTEN group comparing with miR-19 mimic group (P<0.01). Conclusion: miR-19 may activate PI3K/Akt signal pathway by inhibiting PTEN expression, thereby promoting migration and invasion of endometrial cancer KLE cells.
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Project Supported:
Project supported by the Youth Science and Technology Project of Scientific Research Foundation from the Department of Health of Hebei Province (No. 20200126)
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