姜黄素调控唾液腺肿瘤细胞增殖和凋亡的机制探讨

目的: 探讨姜黄素靶向miR-155-5p/TP53INP1轴诱导氧化应激调控唾液腺肿瘤细胞增殖和凋亡的机制。方法: A253细胞加入姜黄素及转染miR-155-5p mimic和（或）pcDNA3.1-TP53INP1,共同培养。采用CCK-8法检测细胞增殖;流式细胞术检测细胞凋亡;划痕实验检测细胞迁移能力;双荧光素酶报告实验验证miR-155-5p与TP53INP1之间的靶向关系;qRT-PCR检测细胞miR-155-5p、TP53INP1mRNA的表达;Western印迹法检测细胞TP53INP1、Caspase8、Caspase3、Bcl-2、Bax蛋白表达;ELISA法检测细胞SOD、Gpx、MDA含量。采用SPSS 22.0软件包对数据进行统计学分析。结果: 双荧光素酶报告实验证实,TP53INP1是miR-155-5p的下游靶向调控分子;与DMSO组相比,姜黄素组细胞凋亡、Caspase8、Caspase3、Bax蛋白表达及TP53INP1表达显著增加,Bcl-2蛋白表达、miR-155-5p mRNA及划痕迁移细胞数显著降低（P<0.05）;与姜黄素+miR-155-5p mimic组相比,姜黄素+pcDNA3.1-TP53INP1组、姜黄素+miR-155-5p mimic+pcDNA3.1-TP53INP1组细胞凋亡、Caspase8、Caspase3、Bax蛋白表达显著增加,Bcl-2蛋白表达显著升高（P<0.05）,姜黄素+pcDNA3.1-TP53INP1组SOD、GSH-PX活性及划痕迁移细胞数显著降低,MDA含量显著升高（P<0.05）。结论: 姜黄素抑制A253细胞增殖,促进A253细胞凋亡,其机制可能与靶向miR-155-5p/TP53INP1轴诱导氧化应激调控有关。

Curcumin targeting miR-155-5p/TP53INP1axis induced oxidative stress to regulate salivary gland tumor cell proliferation,apoptosis

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.