EFFECT OF ITGA9-AS1ON PROLIFERATION, APOPTOSIS AND MIGRATION OF A549/DDP CELLS BY NEGATIVELY REGULATING MIR-629-5P

Jie Yan^*

Supervision Office, Tianjin Medical College, Tianjin, 300222, China

Objective: To investigate the effect of lncRNA ITGA9-AS1 on proliferation, apoptosis and migration of cisplatin (DDP) resistant lung cancer cells A549/DDP and its mechanism. 

Methods: Using para-carcinoma tissue and lung cancer cell A549 as the control, RT-qPCR was used to detect the expression levels of ITGA9-AS1 and miR-629-5p in lung cancer tissues and A549/DDP cells. A549/DDP cells were divided into pcDNA group, pcDNA-ITGA9-AS1 group, pcDNA-ITGA9-AS1+miR-NC group, and pcDNA-ITGA9-AS1+miR-629-5p group. CCK-8 method and clone formation experiment were used to detect cell proliferation; flow cytometry was used to detect apoptosis; Transwell was used to detect cell migration; and Western Blot was used to detect Ki67, Cleaned-caspase3, Pro-caspase3, N-cadherin, and E-cadherin protein expressions. Dual-luciferase reporter gene assay verified the regulatory relationship between ITGA9-AS1 and miR-629-5p. 

Results: Compared with para-carcinoma tissue, lung cancer tissue had significantly reduced ITGA9-AS1 expression (P<0.05). Compared with A549 cells, A549/DDP cells had significantly reduced ITGA9-AS1expression (P<0.05). Compared with pcDNA group, pcDNA-ITGA9-AS1 group had decreased OD value, clone formation number, migrating cell number and Ki67, N-cadherin and Pro-caspase3 protein expression (P<0.05), but increased apoptosis rate and E-cadherin and Cleaned-caspase3 protein expressions in A549/DDP cells (P<0.05). ITGA9-AS1 negatively regulated miR-629-5p expression in A549/DDP cells. Compared with pcDNA-ITGA9-AS1+miR-NC group, pcDNA-ITGA9-AS1+miR-629-5p group had increased OD value, clone formation number, migrating cell number and Ki67, N-cadherin and Pro-caspase3protein expressions (P<0.05), but decreased apoptosis rate, E-cadherin and Cleared-caspase3 protein expressions (P<0.05) in A549/DDP cells. 

Conclusion: Overexpression of ITGA9-AS1 can negatively regulate miR-629-5p to lower proliferation and migration capability of A549/DDP cells and promote its apoptosis.