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The role of hypoxia-induced circ_0007766 expression in the proliferation, migration and invasion of PCa cells
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Abstract:
[Abstract] Objective: To explore the role of circ_0007766 molecule cyclized from the erb-b2 receptor tyrosine kinase 2 (ERBB2) gene in prostate cancer (PCa) cells and the impact of hypoxia on the regulation of its expression. Methods: The expression of circ_0007766 molecule in PCa cells and tissues was detected by qRT-PCR. The siRNA of circ0007766 was transfected to PCa cells (DU145 cell and PC3 cell) respectively; Colony formation assay, CCK-8 assay and Transwell assay were performed to detect the effect of circ_0007766 knockdown on the colony formation, proliferation, migration, and invasion abilities of PCa cells. Hypoxia cell model of DU145 and PC3 cells were established by hypoxic chamber method, and WB was performed to detect the protein expression of HIF-1α, a key molecule of the hypoxic pathway. qRT-PCR was performed to detect the expression of circ_0007766 molecule in hypoxic cell model, and the protein expression of RNA-binding protein eukaryotic translation initiation factor 4A3 (EIF4A3) was detected by WB. RNA immunoprecipitation (RIP) was performed to detect the binding of EIF4A3 to circ_0007766 under hypoxic conditions. qRT-PCR assay was performed to further detect the effect of EIF4A3 knockdown on the expression of circ_0007766 under hypoxic conditions. Results: circ_0007766 was highly expressed in PCa cells (P < 0.01) and PCa tissues (P < 0.05). The knockdown of circ_0007766 (P < 0.05 or P < 0.01) could significantly inhibit the proliferation, migration and invasion abilities of PCa cells (all P < 0.01). The upregulation of HIF-1α protein under hypoxic conditions confirmed the successful establishment of the hypoxia cell model. qRT-PCR analysis revealed that compared with that in the normoxia group, circ_0007766 expression was markedly elevated in the hypoxia group (P < 0.01). WB analysis demonstrated increased EIF4A3 protein expression in cells of the hypoxia group. RIP assays indicated that circ_0007766 was highly concentrated in the EIF4A3-enriched group (P <0.01). Additionally, qRT-PCR showed that hypoxia significantly boosted circ_0007766 expression, whereas EIF4A3 knockdown notably diminished the hypoxia-induced expression of circ_0007766 (P < 0.05 or P < 0.01). Conclusion: Circ-0007766 plays the role of cancer-promoting molecule in PCa cells, and its expression is related to the regulation of EIF4A3 molecule under hypoxic conditions.
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