CircularRNA abhydrolase domain containing 16A-microRNA-1237-5p-apoptosis associated tyrosine kinase axis regulates the apoptosis of michigan cancer foundation-7 in breast cancer cells

Abstract

Objective To investigate the circular RNA abhydrolase domain containing 16A (circRNA ABHD16A)-microRNA (miRNA, miR)-1237-5p-Apoptosis associated tyrosine kinase (AATK) axis on apoptosis of breast cancer cell munggan cancer foundation-7 (MCF-7). Methods The correlation between ABHD16A and miR-1237-5p and miR-1237-5p and AATK was analyzed by CSCD and TargetScan online analysis. The luciferase assay was used to detect whether circRNA ABHD16A targets miR-1237-5p and whether miR-1237-5p targets AATK. The expression of apoptotic marker proteins in AATK and breast cancer cells MCF-7 was detected by Western blot after over-expression or knockdown of circRNA ABHD16A. Cell viability of breast cancer cells MCF-7 was analyzed by methyl thiazolyl tetrazolium (MTT) assay; staining by Annexin-V was performed by MTT assay. The level of apoptosis of breast cancer cells MCF-7 was measured. Student’s t-test analyzed two sets of one-way analysis of variance. Results circRNA ABHD16A targets miR-1237-5p; miR-1237-5p targets the 320-326 and 469-475 regions of the AATK 3’ UTR. When overexpressing ABHD16A, AATK expression increased significantly (t=15.295, P<0.01), apoptosis-related protein bcl-2 antagonist/killer 1 (bak), bcl-2 Associated X Apoptosis Regulator (bax) and cleaved Caspase-9 (cleaved-CAS9) expression levels increased significantly (t=7.403, 8.381, 21.583, P<0.01). The expression of bcl-2 Apoptosis Regulator (bcl-2) was significantly decreased (t=6.301, P<0.01), the activity of MCF7 cells was decreased (t=17.392, P<0.01), and the level of apoptosis was increased (t=8.491, P<0.05). When ABHD16A was knocked down, the expression of AATK was significantly decreased (t=9.290, P<0.01), and the expression levels of apoptosis-related proteins bak, bax and cleaved-CAS9 were significantly decreased (t=8.381, 5.296, 11.492, P<0.01), while the expression of bcl-2 increased significantly (t=5.609, P<0.01), the activity level of MCF7 cells increased (t=5.694, P<0.01), and the level of apoptosis decreased (t=13.502, P<0.01). Conclusion circRNA ABHD16A targets miR-1237-5p and increases the expression of AATK and promotes the apoptosis of breast cancer cells MCF-7. Key words: CircularRNA abhydrolase domain containing 16A; MicroRNA-1237-5p; Apoptosis associated tyrosine kinase; Breast cancer; Michigan cancer foundation-7; Cell apoptosis