二甲双胍和紫草素对人鼻咽癌细胞CNE-2Z增殖及凋亡的影响及其机制  

Effect of metformin in combination with shikonin on CNE-2Z proliferation and apoptosis in nasopharyngeal carcinoma cells and its mechanism

作　　者：王楠楠 苏凯 崔忆旋 陶露 时宗芬 陈晓芬 翟丽倩 马士崟[1] 赵报 WANG Nannan;SU Kai;CUI Yixuan;TAO Lu;SHI Zongfen;CHEN Xiaofen;ZHAI Liqian;MA Shiyin;ZHAO Bao(Department of Otolaryngology,First Affiliated Hospital,Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院第一附属医院耳鼻咽喉科,蚌埠233000

出　　处：《山西医科大学学报》2020年第8期754-760,共7页Journal of Shanxi Medical University

基　　金：蚌埠医学院科技发展基金资助项目(BYKF1763);蚌埠医学院自然科学基金重点项目(BYKY1855ZD);安徽省高校自然科学研究重点项目(KJ2019A0293);国家自然科学基金青年科学基金项目(81903142);安徽省重点研究与开发计划项目(202004j07020007);蚌埠医学院第一附属医院优秀青年科学基金项目(2019byyfyyq03);蚌埠医学院自然科学基金转化医学项目(BYTM2019010)。

摘　　要：目的探讨二甲双胍和紫草素对人鼻咽癌细胞CNE-2Z增殖及凋亡的影响。方法实验共分为4组:空白组、二甲双胍组、紫草素组和二甲双胍+紫草素组。MTT法检测CNE-2Z细胞增殖;AnnexinⅤFITC/PI双染法检测CNE-2Z细胞凋亡;侵袭迁移实验检测细胞侵袭和迁移能力,集落形成实验检测细胞集落形成能力,Western blot法检测CNE-2Z细胞Bcl-2、Bax、MMP9、RIP1、RIP3的表达水平。结果MTT结果示,二甲双胍+紫草素组细胞增殖抑制率较强,且细胞存活率低于二甲双胍组和紫草素组(P<0.05)。AnnexinⅤFITC/PI双染结果示,二甲双胍+紫草素组诱导CNE-2Z细胞凋亡较二甲双胍组、紫草素组更为明显(P<0.05)。侵袭迁移和集落克隆实验结果示,二甲双胍+紫草素组细胞侵袭迁移能力及集落形成能力均明显低于二甲双胍组和紫草素组(P<0.05)。Western blot实验结果示,二甲双胍+紫草素组Bcl-2/Bax比值及MMP9表达明显低于其他三组;RIP1、RIP3表达较二甲双胍组、紫草素组明显上调(P<0.05)。结论二甲双胍和紫草素可能通过诱导坏死性凋亡、促进CNE-2Z细胞凋亡以及下调MMP9表达来抑制CNE-2Z细胞侵袭转移的能力。Objective To investigate the effects of metformin and shikonin on the proliferation and apoptosis of human nasopharyngeal carcinoma cell line CNE-2Z.Methods CNE-2Z cells were divided into four groups:blank group,metformin group,shikonin group and metformin+shikonin group.The proliferation of CNE-2Z cells was detected by MTT assay,and the apoptosis of CNE-2Z cells was detected by AnnexinⅤFITC/PI double staining.The abilities of invasion and migration were detected using invasion and migration assay,and the colony-forming ability was detected by colony formation test.The expression levels of Bcl-2,Bax,MMP9,RIP1 and RIP3 in CNE-2Z cells were detected using Western blot.Results MTT results showed that the inhibitory rate of cell proliferation was lower in metformin+shikonin group than in metformin group and shikonin group,and the cell survival rate was also significantly lower(P<0.05).AnnexinⅤFITC/PI double staining demonstrated that the apoptosis of CNE-2Z cells in metformin+shikonin group was significantly higher than that in metformin group and shikonin group(P<0.05).The abilities of invasion,migration and colony forming in metformin+shikonin group were significantly lower than those in metformin group and shikonin group(P<0.05).Western blot revealed that Bcl-2/Bax ratio and MMP9 expression in metformin+shikonin group were significantly lower than those in metformin group and shikonin group(P<0.05),while RIP1 and RIP3 expression levels were significantly higher(P<0.05).Conclusion Metformin and shikonin may inhibit the invasion and metastasis of CNE-2Z cells by inducing necroptosis,promoting apoptosis of CNE-2Z cells and down-regulating MMP9 expression.

关 键 词：二甲双胍 紫草素 鼻咽癌 坏死性凋亡 

分 类 号：R739.6[医药卫生—肿瘤]