Effects of LncRNA PRR34-AS1 on the Proliferation, Migration, and Invasion of Colorectal Cancer Cells by Regulating the miR-296-5p/DDI2 Axis
CHEN Haiyang, WANG Chunmei*, SHI Jinsheng, DAI Heyang
This study aims to investigate the effect of LncRNA PRR34-AS1 on the proliferation, migration, and invasion of CRC (colorectal cancer) cells by regulating the miR-296-5p/DDI2 axis. qRT-PCR was used to detect LncRNA PRR34-AS1, miR-296-5p, and DDI2 mRNA levels in CRC cells and tissues. Dual luciferase assay was applied to detect the targeting relationships between LncRNA PRR34-AS1 and miR-296-5p, and between DDI2 and miR-296-5p. HCT116 cells were assigned into Control group, sh-NC group, sh-PRR34-AS1 group, sh- PRR34-AS1+anti-NC group, and sh-PRR34-AS1+anti-miR-296-5p group. Except for the Control group, all other groups were transfected with plasmids. The proliferation, migration and invasion of HCT116 cells were detected by cloning method, scratch method and Transwell method, respectively. The expression levels of related proteins were detected by Western blot and immunohistochemistry. The tumor formation of nude mice was applied to detect the effect of LncRNA PRR34-AS1 on the growth of CRC transplanted tumors and on the expression of miR-296-5p and DDI2. In CRC cells and tissues, the expression of LncRNA PRR34-AS1 and DDI2 mRNA increased, while the expression of miR-296-5p decreased (P<0.05). Online prediction showed that LncRNA PRR34-AS1 had a targeted relationship with miR-296-5p, and DDI2 had a targeted relationship with miR-296-5p. Compared with the sh-NC group, the LncRNA PRR34-AS1, DDI2, PCNA, MMP-2, and MMP-9 in the sh-PRR34-AS1 group were downregulated, while miR-296-5p was upregulated, the number of clones formed, scratch healing rate, and number of invasive cells reduced (P<0.05). Compared with the sh-PRR34-AS1+anti-NC group, the miR-296-5p was downregulated in the sh-PRR34-AS1+anti-miR-296-5p group, DDI2, PCNA, MMP-2, MMP-9 were upregulated, and the number of clones formed, scratch healing rate, and number of invasive cells increased (P<0.05). Compared with the sh-NC group, the sh-PRR34-AS1 group had smaller tumor volume and lower mass, the LncRNA PRR34-AS1 and DDI2 were downregulated, while miR-296-5p was upregulated (P<0.05). Inhibition of LncRNA PRR34-AS1 can target the miR-296-5p/DDI2 axis to inhibit the proliferation, migration, and invasion of CRC cells.
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