丹参酮ⅡA通过miR-29a/LC3信号通路抑制鼻咽癌CNE2细胞增殖  

TanshinoneⅡA inhibits the proliferation of nasopharyngeal carcinoma CNE2 cells by regulating miR-29a/LC3 signal pathway

作　　者：王有虎[1] 靳文瑶 王俊博 席克虎[1] 乔自林[3] WANG You-hu;JIN Wen-yao;WANG Jun-bo;XI Ke-hu;QIAO Zi-lin(Department of Otorhinolaryngology Head and Neck Surgery,the First Hospital of Lanzhou University,Lanzhou 730000,Gansu,China;Department of Anatomy and Embryology,Peking University Basic Medicine School,Beijing 100191,China;Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,Gansu,China)

机构地区：[1]兰州大学第一医院耳鼻咽喉头颈外科,甘肃兰州730000 [2]北京大学基础医学院人体解剖与组织胚胎学系,北京100191 [3]西北民族大学生物医学研究中心,甘肃兰州730030

出　　处：《西北师范大学学报（自然科学版）》2021年第6期75-80,87,共7页Journal of Northwest Normal University(Natural Science)

基　　金：教育部创新性团队发展计划项目(IRT-17R88);中央高校基本科研业务费专项资金资助项目(31920190004);甘肃省自然科学基金资助项目(21JRTRA363)。

摘　　要：探究丹参酮ⅡA对鼻咽癌CNE2细胞增殖的影响,并研究其影响的具体机制.通过MTT法检测不同浓度及不同作用时间下丹参酮ⅡA对CNE2细胞增殖的抑制情况并计算细胞增殖率,采用caspase3及caspase9活性试剂盒评估丹参酮ⅡA对CNE2细胞凋亡的影响,利用RT-PCR及Western Blot技术检测了自噬相关分子LC3的表达,并用miR-29a拮抗剂探索miR-29a在CNE2细胞增殖过程中的作用.结果表明,丹参酮ⅡA对鼻咽癌CNE2细胞增殖具有显著的抑制作用(P<0.05),且抑制作用呈剂量和时间依赖性;与对照组相比,4及8μmol·L^(-1)的丹参酮ⅡA作用于CNE2细胞72 h后,细胞内caspase3及caspase9活性均显著升高(P<0.05);丹参酮ⅡA作用后,细胞自噬相关分子LC3表达水平显著升高,丹参酮ⅡA可提高CNE2细胞内miR-29a的表达水平;在CNE2细胞内转染miR-29a拮抗剂后,丹参酮ⅡA对CNE2细胞增殖的抑制作用降低,抑制miR-29a亦可降低CNE2细胞内LC3表达水平.丹参酮ⅡA通过上调miR-29a的表达,增强CNE2细胞自噬及凋亡水平,抑制鼻咽癌CNE2细胞增殖,这为临床治疗鼻咽癌提供了新的潜在药物.In this paper,the effect of tanshinoneⅡA on the proliferation of CNE2 cells was investigated and the underlying mechanism was explored.After treatment of tanshinoneⅡA in different concentration and duration,the cell proliferation of CNE2 was detected by MTT assay,the proliferation rate was calculated,the caspase3 and caspase9 activity kit were used to assess the effect of tanshinoneⅡA on the apoptosis,the expression level of LC3 was detected by using real-time PCR and Western Blot,and the role of miR-29a was accessed by transfecting miR-29 antagonist into CNE2 cells after tanshinoneⅡA treatment.The results showed that administration of tanshinoneⅡA significantly inhibits the proliferation of CNE2 cells,which is dependent on the concentration and duration(P<0.05).Compared with the negative control,treatment of tanshinoneⅡA can lead to an increasing activity of caspase3 as well as caspase9 in the concentration of 4 and 8μmol·L^(-1)(P<0.05).After tanshinoneⅡA administration,the expression of LC3 is obviously increased(P<0.05).Notably,tanshinoneⅡA treatment can significantly upregulate the expression of miR-29a.After transfection of miR-29a antagonist,the proliferation of CNE2 cells is significantly upregulated compared to the controls.Moreover,miR-29a antagonist can remarkedly reduce the expression of LC3 when exposes to tanshinoneⅡA.TanshinoneⅡA can enhance the pyroptosis and apoptosis on CNE2 cells by regulating miR-29a/LC3 signal pathway and lead to an inhibition of the cell proliferation,which can provide a potential drug for the clinic therapy of nasopharyngeal carcinoma.

关 键 词：丹参酮ⅡA 鼻咽癌 细胞增殖 自噬 miR-29a 

分 类 号：R285[医药卫生—中药学]