甲硫氨酸限制对口腔癌细胞增殖、迁移及侵袭的影响  

Effects of methionine restriction on oral cancer cell proliferation,migration and invasion

作　　者：彭月荣 郑可欣 陈涵思 王学鹏 赖运浩 周素瑾 赵子建 赵正刚 李芳红 PENG Yue-rong;ZHENG Ke-xin;CHEN Han-si;WANG Xue-peng;LAI Yun-hao;ZHOU Su-jin;ZHAO Zi-jian;ZHAO Zheng-gang;LI Fang-hong(School of Biomedical and Pharmaceutical Sciences,Guangdong University of Technology,Guangzhou 510006,China)

机构地区：[1]广东工业大学生物医药学院,广东广州510006

出　　处：《中国药理学通报》2023年第8期1444-1450,共7页Chinese Pharmacological Bulletin

基　　金：国家重点研发计划(No 2018YFA0800603);广东省重点领域研发计划“新药创制”专项(No 2019B020201015);广东省创新团队(No 2016ZT06Y432);广州市基础与应用基础研究基金(No 202102021117,202102020159)。

摘　　要：目的探讨甲硫氨酸限制对人口腔鳞状癌细胞CAL-27的增殖、迁移和侵袭的影响。方法采用计数法检测甲硫氨酸限制对口腔癌细胞CAL-27增殖的影响;平板克隆法检测CAL-27细胞克隆形成;流式细胞术结合PI单染法检测CAL-27细胞的周期;Annexin V-PE/7AAD双染法检测CAL-27细胞的凋亡;划痕与Transwell实验检测CAL-27细胞的迁移和侵袭能力;Western blot检测细胞凋亡蛋白Bcl-2和Bax、细胞周期蛋白CDK2和CDK4以及迁移侵袭蛋白N-cadherin和E-cadherin的表达水平的变化。结果甲硫氨酸限制明显抑制口腔癌CAL-27细胞的增殖和克隆形成(P<0.01);甲硫氨酸限制将口腔癌细胞CAL-27阻滞在G_(0)/G_(1)期(P<0.01);甲硫氨酸限制明显诱导口腔癌细胞CAL-27细胞发生凋亡,抑制细胞的迁移和侵袭能力(P<0.01);Western blot结果表明甲硫氨酸限制明显下调口腔癌CAL-27细胞的凋亡蛋白Bcl-2的表达以及周期蛋白CDK2和CDK4的表达,并且明显下调迁移和侵袭蛋白N-cadherin以及上调E-cadherin的表达水平。结论口腔癌细胞CAL-27具有甲硫氨酸依赖性;通过甲硫氨酸限制明显抑制口腔癌细胞CAL-27细胞的增殖、迁移和侵袭,可为甲硫氨酸限制疗法应用于口腔癌的治疗提供理论依据。Aim To investigate the effect of methionine restriction on the proliferation,migration and invasion of human oral squamous carcinoma CAL-27 cells.Methods Cell proliferation and colony formation ability were detected by cell counting and colony forming assay.The changes in cell cycle and apoptosis were detected by propidium iodide(PI)staining flow cytometry and Annexin V/7-amino-actinomycin staining flow cytometry.The migration and invasion ability of CAL-27 was detected by scratch and Transwell assay.The expression levels of apoptosis proteins Bax and Bcl-2,cyclins CDK2 and CDK4 and migration and invasion proteins N-cadherin and E-cadherin were examined by Western blot.Results Methionine restriction significantly inhibited the proliferation and clone formation of oral squamous cancer cell CAL-27(P<0.01),induced cell cycle arrest at G_(0)/G_(1) phase(P<0.01)and significantly induced apoptosis.Besides,methionine restriction inhibited cell migration and invasion(P<0.01).Western blot results showed that methionine restriction significantly down-regulated the expression of apoptosis proteins Bcl-2,cyclins CDK2 and CDK4,and also significantly decreased the expression of migration and invasion proteins N-cadherin and increased the expression of E-cadherin in oral cancer CAL-27 cells.Conclusions Oral cancer cell CAL-27 is methionine-dependent,and methionine restriction significantly inhibits the proliferation,migration and invasion of oral cancer cell CAL-27 cells,which may provide a new theoretical basis for methionine restriction therapy with oral cancer.

关 键 词：口腔癌 甲硫氨酸限制 增殖 细胞周期 侵袭 迁移 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R739.8[医药卫生—基础医学] R977.4R979.1