β-Lapachone通过诱导ROS生成调控口腔鳞状细胞癌增殖与凋亡的机制研究

doi:10.13701/j.cnki.kqyxyj.2022.01.009

摘要/Abstract

摘要： 目的:探明NQO1蛋白在口腔鳞状细胞癌(OSCC)组织中的表达,研究NQO1生物可激活药物β-拉帕醌(β-Lapachone)体外抑制OSCC发生与发展的分子机制。方法:(1)免疫组织化学染色法(IHC)检测50例OSCC组织和9例正常口腔黏膜组织中NQO1蛋白的表达,分析NQO1表达与OSCC患者病理特征之间的关系;(2)体外常规培养HN22和HSC4细胞。采用CCK-8、平板克隆实验检测β-Lapachone对OSCC细胞增殖的抑制作用。流式细胞术检测β-Lapachone在OSCC细胞凋亡中作用;(3)β-Lapachone处理后,检测OSCC细胞活性氧(ROS)生成。在ROS抑制剂NAC(N-acetyl-L-cysteine)干预下,检测β-Lapachone对 OSCC细胞的抗增殖能力是否有影响。结果:(1)免疫组织化学结果显示NQO1蛋白在OSCC中表达阳性率和强阳性率显著高于正常组织(P<0.05);χ²检验结果显示,NQO1蛋白表达水平与OSCC患者肿瘤大小、病理分级密切相关(P值分别为0.007和0.041);(2)CCK-8、平板克隆、Edu实验结果显示β-Lapachone可抑制OSCC的增殖能力。流式细胞术检测结果显示β-Lapachone诱导OSCC细胞凋亡;(3)DCFH-DA检测结果显示β-Lapachone可诱导ROS的生成。 NAC可部分逆转β-Lapachone在OSCC细胞中的抗增殖作用。结论:NQO1蛋白与OSCC的发生发展密切相关,且其生物激活剂β-Lapachone通过诱导ROS生成,调控OSCC细胞的增殖和凋亡。

关键词: 口腔鳞状细胞癌, NQO1, β-拉帕醌, 活性氧, 细胞增殖, 细胞凋亡

Abstract: Objective: To evaluate the expression level of NQO1 in OSCC, and investigate the effect of β-Lapachone on proliferation, apoptosis, and ROS generation in OSCC. Methods: The expression of NQO1 was detected by IHC staining in 50 cases of OSCC tissues and 20 cases of normal oral tissues. Cell viability was detected using CCK-8 and colony formation and Edu incorporation assay after β-Lapachone treatment. The effect of β-Lapachone on cell apoptosis was detected by flowcytometry. ROS generation was determined by DCFH-DA method. ROS inhibitor (NAC) was applied to determine the effect of ROS generation on OSCC cell proliferation. Results: IHC staining showed that the positive and strongly positive rate of NQO1 was significantly higher than that of normal tissues (P<0.05). The expression of NQO1 was correlated with tumor size and histological grade. The results of CCK-8 and colony formation and Edu incorporation assay showed that β-Lapachone suppressed OSCC cell proliferation. In addition, Annexin V/7-AAD staining showed that β-Lapachone induced OSCC cell apoptosis. Additionally, β-Lapachone could induce ROS generation. ROS inhibitor (NAC) could partially attenuate the effect of β-Lapachone on cell proliferation. Conclusion: The expression of NQO1 was correlated with OSCC development and progression. β-Lapachone regulated cell proliferation and apoptosis through regulating ROS generation.

Key words: OSCC, NQO1, β-Lapachone, ROS, proliferation, apoptosis

林晶, 金成日, 玄云泽. β-Lapachone通过诱导ROS生成调控口腔鳞状细胞癌增殖与凋亡的机制研究[J]. 口腔医学研究, 2022, 38(1): 39-44.

LIN Jing, JIN Chengri, XUAN Yunze. β-Lapachone Suppresses OSCC Proliferation and Induces Cell Apoptosis through ROS Generation[J]. Journal of Oral Science Research, 2022, 38(1): 39-44.

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