Search

Objective: To investigate the regulatory mechanism of miR-216a-5p on autophagy and radiosensitivity of gastric cancer cell and its regulatory effect on dual-specificity phosphatase 10 (DUSP10). Methods: SGC-7901 cells were irradiated with linear accelerator 6-MV X-ray at a dose rate of 0.8 Gy/min. The total dose is 8Gy. miR-216a-5p-mimic, NC-mimic, pcDNA-DUSP10 or pcDNA-NC were transfected into SGC-7901 cells with Lipofectamine 2000 reagent. After transfection, the cells were divided into miR-216a-5p-mimic group and NC-mimic group, and each group was divided into two subgroups, 0Gy and 8Gy. In the rescue experiment, the cells were divided into miR-216a-5p mimic+pcDNA DUSP10 group and miR-216a-5p mimic+pcDNA NC group. The mRNA levels of miR-216a-5p and DUSP10 were detected by qRT-PCR. Cell proliferation was detected by 5-ethynyl-2'-deoxyuridine (EdU) incorporation test and colony formation assay. The apoptosis was assessed by flow cytometry. The protein expressions of DUSP10, Bax, Bad, Bcl-2, LC3 and p62 were detected by Western blot. The expression of γH2AX was detected by immunofluorescence method to evaluate DNA double-strand break (DSB) in cells. The targeting relationship between miR-216a-5p and DUSP10 was detected by luciferase reporter gene. Autophagosomes were detected by GFP-mRFP-LC3. Results: Compared with the 8Gy NC-mimic group, the number of colonies, EdU positive rate and Bcl-2 protein expression level in the 8Gy miR-216a-5p-mimic group decreased, while the positive rate of γH2AX, the rate of apoptosis and the expression levels of Bax and Bad proteins increased (P<0.01). Compared with the 8Gy NC-mimic group, the number of autophagosomes and the expression level of LC3II protein in the 8Gy miR-216a-5p-mimic group decreased, while the expression level of p62 protein increased (P<0.001). After co-culture with miR-216a-5p-mimic, the relative luciferase activity of DUSP10-3'-UTR-WT was significantly reduced compared with the DUSP10-3'-UTR-MUT group (P<0.001). Compared with the NC-mimic group, the DUSP10 mRNA and protein expression levels in the miR-216a-5p-mimic group were lower (P<0.001). Compared with the miR-216a-5p mimic+pcDNA NC group, the number of colonies and autophagosomes in the miR-216a-5p mimic+pcDNA DUSP10 group increased, while the apoptosis rate was decreased (P<0.001). Conclusion: miR-216a-5p inhibits cell proliferation, increases radiation-induced apoptosis and inhibits radiation-induced autophagy by inhibiting DUSP10, thereby enhancing the radiosensitivity of gastric cancer cells. [ABSTRACT FROM AUTHOR]

Copyright of Progress in Modern Biomedicine is the property of Publishing House of Progress in Modern Biomedicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Progress in Modern Biomedicine is the property of Publishing House of Progress in Modern Biomedicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)