MiR-195-5p对食管鳞癌细胞放射敏感性的影响及作用机制研究

doi:10.3969/j.issn.1004-616x.2021.02.001

Abstract

Abstract: OBJECTIVE: To investigate effects and mechanism of miR-195-5p on radiosensitivity of esophageal squamous cell carcinoma cells in vitro. METHODS: KYSE510 and KYSE150 cells were exposed to an 8 Gy of X-rays,and non-irradiated cells were used as control. mRNA expression levels of miR-195-5p and of survivin were detected by qPCR. Protein expression levels of survivin were detected by Western blot. KYSE510 cells were transfected with different oligos which gave rise to the miR-195-5p mimic,antagomir and negative control groups. After KYSE510 cells were irradiated with X-rays,cell proliferation was measured by the Incucyte real-time dynamic living cell monitoring and EdU incorporation assays. Cell colony survival and apoptosis rates were measured by colony formation assay and flow cytometry analysis,respectively. Dual-luciferase reporter assay was used to verify the luciferase activity of KYSE150 cells in each group. RESULTS: Growth of KYSE510 and KYSE150 cells were significantly inhibited after irradiation. Compared with the non-irradiation group,expression of miR-195-5p in the irradiation group was decreased,and expression of survivin protein was increased (P<0.05 or 0.01). After X-ray irradiation,the survival rate of the miR-195-5p mimic group was lower than that of the negative control group (P<0.01). The rates for EdU positive cells and clonal survival were lower than that of the negative control group (P<0.05 or 0.01),and the apoptosis rate was significantly higher than that of the negative control group (P<0.01). However,the rates for EdU positive cells and clonal survival of the antagomir group was higher than that of the negative control group (P<0.01). The expressions of survivin mRNA and protein in the miR-195-5p mimic group was lower than that of the negative control group. In contrast,the expressions of survivin mRNA and protein in the miR-195-5p antagomir group was higher than that of the negative control group. Over-expression of miR-195-5p in KYSE150 cells inhibited luciferase activity of the BIRC5 3' UTR reporter gene (P<0.05). CONCLUSION: MiR-195-5p could enhance radiosensitivity of esophageal squamous cell carcinoma cells in vitro by targeting survivin gene.

Key words: miR-195-5p, survivin, radiosensitivity, esophageal squamous cell carcinoma

CLC Number:

R730.5

ZHANG Die, LIU Xianghe, HUO Miaomiao, LIU Mei, XU Ningzhi, ZHU Hongxia. Effects of miR-195-5p on radiosensitivity of esophageal squamous cell carcinoma cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(2): 81-88,94.

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Effects of miR-195-5p on radiosensitivity of esophageal squamous cell carcinoma cells

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