DU Hong-Liang, He Deng-Qi, Wang Jia-Li, Che Yin-Fu, Yan Qing-Han
Department of Oral and Maxillofacial Surgery, The First Hospital of Lanzhou University. Lanzhou 730000, Gansu Province, China. E-mail:
Shanghai Kou Qiang Yi Xue. 2022 Oct;31(5):483-490.
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.
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.
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).
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.
探讨姜黄素靶向miR-155-5p/TP53INP1轴诱导氧化应激以调控涎腺肿瘤细胞增殖和凋亡的机制。
培养A253细胞，加入姜黄素并转染miR-155-5p模拟物和/或pcDNA3.1-TP53INP1。采用CCK-8法检测细胞增殖，流式细胞术检测细胞凋亡，划痕试验检测细胞迁移能力。通过双荧光素酶报告基因检测法验证miR-155-5p与TP53INP1的靶向关系。采用qRT-PCR检测miR-155-5p、TP53INP1 mRNA表达。通过蛋白质免疫印迹法检测TP53INP1、Caspase8、Caspase3、Bcl-2、Bax蛋白表达；采用酶联免疫吸附测定法测定超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（Gpx）和丙二醛（MDA）含量。使用SPSS 22.0软件包进行统计分析。
双荧光素酶报告基因检测法证实TP53INP1是miR-155-5p的下游靶标调控分子。与二甲基亚砜（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，而Bcl-2蛋白表达、miR-155-5p mRNA及细胞迁移数量显著减少（P<０.０５）。与姜黄素＋miR-155-5p模拟物组相比，姜黄素＋pcDNA3.1-TP53INP1组和姜黄素＋miR-155-5p模拟物＋pcDNA3.1-TP53INP1组细胞凋亡、Caspase8、Caspase3、Bax蛋白表达显著增加；Bcl-2蛋白表达显著增加（P<０.０５），姜黄素＋pcDNA3.1-TP53INP1组超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-PX）活性及细胞迁移数量显著减少，丙二醛（MDA）含量显著增加（P<０.０５）。
姜黄素抑制A253细胞增殖并促进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。
