MiR-103a-3p Contributes to the Progression of Colorectal Cancer by Regulating GREM2 Expression
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Purpose: Our research aimed to investigate the influence of miR-103a-3p on the growth and apoptosis of colorectal cancer (CRC)cells. Materials and Methods: Bioinformatics was employed to analyze differentially expressed microRNAs and predict target genes. qRT-PCR was applied to detect the expression of miR-103a-3p in CRC and normal cells. HCT116 and Caco-2 were chosen, andmiR-103a-3p mimics, miR-103a-3p inhibitor, as well as specific siRNAs targeting GREM2, were constructed. We subsequentlyevaluated alternations in cell proliferation, cell cycle and cell cycle regulators, apoptosis, and related proteins (Bcl-2 and Bax) byCCK-8 testing, Western blotting, luciferase reporter, colony formation, and Annexin V-FITC/PI. Possible binding sites for miR-103a-3p on the 3'UTR of GREM2 were checked with luciferase assay, and the impact of GREM2 on miR-103a-3p activity was also validatedwith above biological function testing. Additionally, the effect of miR-103a-3p knockdown in CRC cells and the molecularmechanism of miR-103a-3p targeting GREM2 were also studied. Results: Bioinformatics analysis revealed that miR-103a-3p expression increased remarkably in CRC, and targeted regulatory correlationexisted between miR-103a-3p and GREM2. MiR-103a-3p inhibitor significantly impeded proliferative capacity and caused cellcycle arrest, as well as apoptosis, in HCT116 and Caco-2 cells. Consistent with this finding, overexpression of GREM2 showed similareffects to miR-103a-3p inhibition. Moreover, we demonstrated that miR-103a-3p connected target GREM2 and GREM2 knockdownreversed the effects of miR-103a-3p inhibitor on HCT116 and Caco-2 cell proliferation, cell cycle, and apoptosis. Further study showedthat miR-103a-3p targeting GREM2 appeared to affect CRC progression via the transforming growth factor-β pathway. Conclusion: MiR-103a-3p could augment CRC progression by targeting GREM2 and that miR-103a-3p/GREM2 could be potentialnovel targets for CRC therapy.
Abstract
Purpose: Our research aimed to investigate the influence of miR-103a-3p on the growth and apoptosis of colorectal cancer (CRC)cells. Materials and Methods: Bioinformatics was employed to analyze differentially expressed microRNAs and predict target genes. qRT-PCR was applied to detect the expression of miR-103a-3p in CRC and normal cells. HCT116 and Caco-2 were chosen, andmiR-103a-3p mimics, miR-103a-3p inhibitor, as well as specific siRNAs targeting GREM2, were constructed. We subsequentlyevaluated alternations in cell proliferation, cell cycle and cell cycle regulators, apoptosis, and related proteins (Bcl-2 and Bax) byCCK-8 testing, Western blotting, luciferase reporter, colony formation, and Annexin V-FITC/PI. Possible binding sites for miR-103a-3p on the 3'UTR of GREM2 were checked with luciferase assay, and the impact of GREM2 on miR-103a-3p activity was also validatedwith above biological function testing. Additionally, the effect of miR-103a-3p knockdown in CRC cells and the molecularmechanism of miR-103a-3p targeting GREM2 were also studied. Results: Bioinformatics analysis revealed that miR-103a-3p expression increased remarkably in CRC, and targeted regulatory correlationexisted between miR-103a-3p and GREM2. MiR-103a-3p inhibitor significantly impeded proliferative capacity and caused cellcycle arrest, as well as apoptosis, in HCT116 and Caco-2 cells. Consistent with this finding, overexpression of GREM2 showed similareffects to miR-103a-3p inhibition. Moreover, we demonstrated that miR-103a-3p connected target GREM2 and GREM2 knockdownreversed the effects of miR-103a-3p inhibitor on HCT116 and Caco-2 cell proliferation, cell cycle, and apoptosis. Further study showedthat miR-103a-3p targeting GREM2 appeared to affect CRC progression via the transforming growth factor-β pathway. Conclusion: MiR-103a-3p could augment CRC progression by targeting GREM2 and that miR-103a-3p/GREM2 could be potentialnovel targets for CRC therapy.
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