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[Curcumin targeting miR-155-5p/TP53INP1 axis induced oxidative stress to regulate salivary gland tumor cell
Hong-Liang DU1, Deng-Qi He, Jia-Li Wang
1Department of Oral and Maxillofacial Surgery, The First Hospital of Lanzhou University. Lanzhou 730000, Gansu Province, China.
Purpose:
To investigate the mechanism of curcumin targeting miR-155-5p/TP53INP1 axis to induce oxidative stress to regulate salivary gland tumor cell proliferation and apoptosis.
Methods:
A253 cells were cultured by adding curcumin and transfected with miR-155-5p mimic and/or pcDNA3.1-TP53INP1. Cell proliferation was detected by CCK-8 assay cell apoptosis was detected by flow cytometry, cell migration ability was detected by scratch test. The targeting relationship between miR-155-5p and TP53INP1 was verified by dual luciferase reporter assay. miR-155-5p, TP53INP1 mRNA expression was detected by qRT-PCR. Western blot was performed to detect expression of TP53INP1, Caspase8, Caspase3, Bcl-2, Bax protein; and ELISA was used to determine SOD, Gpx, and MDA content. Statistical analysis was performed using SPSS 22.0 software package.
Results:
Dual luciferase reporter assay confirmed that TP53INP1 was a downstream target regulatory molecule of miR-155-5p. Compared with DMSO group, cell apoptosis, Caspase8, Caspase3, Bax protein expression and TP53INP1 expression were significantly increased in curcumin group, while Bcl-2 protein expression, miR-155-5p mRNA and number of cell migration were significantly decreased(P<0.05). Compared with curcumin + miR-155-5p mimic group, cell apoptosis, Caspase8, Caspase3, Bax protein expression was significantly increased in curcumin + pcDNA3.1-TP53INP1 group and curcumin + miR-155-5p mimic + pcDNA3.1-TP53INP1 group; Bcl-2 protein expression was significantly increased(P<0.05), SOD, GSH-PX activities and number of cell migration were significantly decreased and MDA content was significantly increased in curcumin+pcDNA3.1-TP53INP1 group (P<0.05).
Conclusions:
Curcumin inhibited A253 cell proliferation and promoted A253 cell apoptosis. The mechanism may be related to targeting miR-155-5p/TP53INP1 axis to induce oxidative stress regulation.
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