Krishnan Manu, Saraswathy Seema, Singh Sanjana, Saggu Gursimran Kaur, Kalra Namita, Agrawala Paban K, Abraham Kurian Mathew, Das Toora Bishamber
Commanding Officer & Classified Specialist (Orthodontics), Military Dental Centre, Ahmednagar, India.
Faculty (Biochemistry), Army College of Medical Sciences (ACMS), Delhi Cantt, India.
Med J Armed Forces India. 2024 Jul-Aug;80(4):412-419. doi: 10.1016/j.mjafi.2022.03.005. Epub 2022 Jun 10.
Biologic compounds have recently generated interest in cancer chemoprevention. Sulforaphane (SFN), an isothiocyanate from cruciferous vegetables, has profound epigenetic actions. Since epigenetic aetiology is crucial for oral cancer, this study evaluated the role of SFN in oral cancer prevention.
Oral squamous cell carcinoma cells (UPCI-SCC-172) were treated with SFN in three concentrations: 10 μM, 20 μM and 30 μM for two time periods: 24 h and 48 h. MTT assay assessed cell proliferation. Histone deacetylase (HDAC) enzyme activity was colorimetrically estimated in the nuclear extracts. Flow cytometry determined cell cycle stages, reactive oxygen species (ROS) generation and mitochondrial membrane potential (MMP) changes. Extrinsic and intrinsic apoptotic pathways were evaluated from caspase enzyme assays.
Cell proliferation and HDAC activity (44% in 24 h and 40% in 48 h) were significantly inhibited (p < 0.01). For 10 μM concentration, G2/M cell cycle arrest was found with a reduction in G1 phase cell population at 24 h and 48 h. Concentrations of 20 μM and 30 μM SFN presented cells in apoptosis marked by increased sub G1 cells at 48 h. Concentrations of 10 μM and 20 μM SFN showed a 1.3 to 2.8-fold increase in ROS generation at 24 h and 48 h. The concentration of 30 μM SFN showed a drop in ROS production, denoting cells already in apoptosis. Fall in MMP was also dose- and time-dependent. Caspase enzyme assays (p < 0.001) demonstrated activation of both extrinsic and intrinsic apoptotic pathways.
Inhibitory action of SFN on oral cancer cell proliferation and HDAC activity led to cell cycle arrest and apoptosis. These effects marked by increase in ROS, a decrease in MMP and activation of apoptotic pathways offer exciting therapeutic options.
生物化合物最近在癌症化学预防方面引起了关注。萝卜硫素（SFN）是一种来自十字花科蔬菜的异硫氰酸盐，具有深远的表观遗传作用。由于表观遗传病因学对口腔癌至关重要，本研究评估了SFN在口腔癌预防中的作用。
用三种浓度（10μM、20μM和30μM）的SFN处理口腔鳞状细胞癌细胞（UPCI-SCC-172），处理两个时间段（24小时和48小时）。MTT法评估细胞增殖。采用比色法测定核提取物中的组蛋白脱乙酰酶（HDAC）活性。流式细胞术确定细胞周期阶段、活性氧（ROS）生成和线粒体膜电位（MMP）变化。通过半胱天冬酶酶分析评估外源性和内源性凋亡途径。
细胞增殖和HDAC活性（24小时时为44%，48小时时为40%）受到显著抑制（p<0.01）。对于10μM浓度，在24小时和48小时时发现G2/M期细胞周期停滞，同时G1期细胞群体减少。20μM和30μM SFN浓度在48小时时使细胞呈现凋亡，表现为亚G1期细胞增加。10μM和20μM SFN浓度在24小时和48小时时使ROS生成增加1.3至2.8倍。30μM SFN浓度显示ROS生成下降，表明细胞已处于凋亡状态。MMP的下降也是剂量和时间依赖性的。半胱天冬酶酶分析（p<0.001）表明外源性和内源性凋亡途径均被激活。
SFN对口腔癌细胞增殖和HDAC活性的抑制作用导致细胞周期停滞和凋亡。这些以ROS增加、MMP降低和凋亡途径激活为特征的效应提供了令人兴奋的治疗选择。
