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Archive > Volume 29 Issue 3 > 2022,29(3):202-208. DOI:10.3872/j.issn.1007-385X.2022.03.006 Prev Next
miR-620 regulates radiosensitivity of breast cancer MCF-7 cells by targeting ING4
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Abstract:
Objective: To investigate the effect of miR-620 on the radiosensitivity of breast cancer MCF-7 cells and its mechanism. Methods: The cancer tissues and para-cancerous tissue specimens of 21 patients with breast cancer who had surgical resection in Danzhou City People's Hospital of Hainan Province from March 2017 to March 2018 were collected. In addition, breast cancer MCF-7 and BCaP-37 cells and breast epithelial HBL-100 cells were also cultured in vitro. The mRNA expression of miR-620 and inhibitor of growth 4 (ING4) in breast cancer tissues and cells was detected by qRT-PCR. MCF-7 cells were respectively transfected with miR-620 inhibitor (anti-miR-620), inhibitor negative control (anti-miR-NC), anti-miR-620 with ING4 small interfering RNA (si-ING4), and anti-miR-620 with small interfering RNA negative control (si-NC) using Lipofectamine TM 2000 lipofection technology before radiotherapy, namely IR+anti-miR-620 group, IR+anti-miR-NC group, IR+anti-miR-620+si-ING4 group, and IR+anti-miR-620+si-NC group, respectively.The cell radiosensitivity, cell proliferation activity, cell cycle distribution and apoptosis rate were detected by clone formation assay, MTT and FCM, respectively. The dual-luciferase reporter gene assay and Western blotting were used to verify the targeting relationship between miR-620 and ING4. Results: Compared with para-cancerous tissues and HBL-100 cells, the expression of miR-620 was significantly increased while the mRNA expression of ING4 was significantly reduced in breast cancer tissues and cells(all P<0.01). Compared with the IR+anti-miR-NC group, the proliferation activity and S phase cell proportion of MCF-7 cells in the IR+anti-miR-620 group were significantly decreased (all P<0.01), but the apoptosis rate, the proportion of G0-G1 phase cells, and the radiosensitivity were significantly increased (all P<0.01). Compared with the IR+anti-miR-620+si-NC group, the proliferation activity and the S-phase cell proportion of MCF-7 cells in the IR+anti-miR-620+si-ING4 group were significantly increased (all P<0.01), but the apoptosis rate, the proportion of G0-G1 phase cells, and the radiosensitivity were significantly decreased (all P<0.01). The dual-luciferase reporter gene assay showed that ING4 is a target gene of miR-620, and miR-620 targeted and negatively regulated the expression of ING4. Conclusion: Knockdown of miR-620 could inhibit the proliferation of MCF-7 cells but promote the apoptosis and radiosensitivity of MCF-7 cells by up-regulating ING4.
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Clc Number:
R737.9；R730.2
