硝呋齐特对甲状腺乳头状癌细胞增殖、迁移及侵袭的影响  

Effect of Nifuroxazide on Proliferation,Migration,and Invasion of Thyroid Papillary Carcinoma Cells

作　　者：胡宇[1] 梁利波[2] 张勤[1] 兰菲[1] 林垦[1] 何訸[3] 李双庆[2] HU Yu;LIANG Li-bo;ZHANG Qin;LAN Fei;LIN Ken;HE He;LI Shuang-qing(Departmant of Endocrinology and Metabolism,Chengdu First People 's Hospital,Montpellier France-Chengdu China Research Institution of Endocrine and Metabolic Diseases,Chengdu 610041,China;Department of General Practice, West China Hospital,Sichuan University,Chengdu 610041,China;Department of Laboratory Medicine,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]成都市第一人民医院内分泌代谢科法国蒙彼利埃-中国成都内分泌代谢疾病研究所,成都610041 [2]四川大学华西医院全科医学科,成都610041 [3]四川大学华西医院实验医学科,成都610041

出　　处：《四川大学学报（医学版）》2019年第1期48-54,共7页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金青年科学基金(No.81500645);四川省国际科技合作与交流研发项目(No.2018HH0065);四川省卫计委科研课题(No.130407)资助

摘　　要：目的探讨硝呋齐特对甲状腺乳头状癌细胞BCPAP和TPC-1的增殖、迁移及侵袭的影响。方法用不同浓度(0、1.25、2.5、5、10、20μmol/L)硝呋齐特处理BCPAP和TPC-1细胞,采用MTT法和克隆形成实验观察硝呋齐特对细胞增殖的影响;采用Hoechst 33258染色及流式分选技术观察对细胞凋亡的影响;通过Western blot法检测硝呋齐特处理后BCPAP细胞凋亡相关蛋白及迁移侵袭相关蛋白的表达情况;采用Transwell小室观察细胞迁移和侵袭能力。结果硝呋齐特0、1.25、2.5μmol/L处理BCPAP细胞,0、1.25μmol/L处理TPC-1细胞,24、48、72h后细胞增殖力均未受到明显抑制(P>0.05);硝呋齐特5、10、20μmol/L处理BCPAP细胞,10、20μmol/L处理TPC-1细胞,24、48、72h后细胞增殖力均受到明显抑制(P<0.05),且随浓度增加和时间延长其抑制作用逐渐增强;此外,2.5、5μmol/L硝呋齐特对TPC-1细胞的增殖抑制作用分别从72h和48h起效(P<0.05)。暴露于10μmol/L硝呋齐特10d后,BCPAP和TPC-1细胞的克隆形成均受到抑制(P<0.05)。10μmol/L硝呋齐特处理48h后,BCPAP和TPC-1细胞均出现细胞核改变,凋亡小体出现;流式分析显示BCPAP及TPC-1凋亡细胞增加(P<0.05)。10μmol/L硝呋齐特处理BCPAP细胞48h,促凋亡蛋白CC-3、Bax的表达增加(P<0.05),而抗凋亡蛋白Bcl-2表达减少(P<0.05);促迁移侵袭相关蛋白基质金属蛋白酶(MMP)-2、MMP-9的表达减少(P<0.05),MMPs家族抑制剂——金属蛋白酶组织抑制剂(TIMP)-2表达增加(P<0.05)。10μmol/L硝呋齐特处理BCPAP和TPC-1细胞48h,BCPAP细胞及TPC-1细胞迁移率与侵袭率均下降(P<0.05)。结论硝呋齐特于体外抑制人甲状腺癌细胞系BCPAP和TPC-1的增殖,通过上调CC-3和Bax蛋白的表达诱导细胞凋亡,通过降低MMP-2和MMP-9蛋白的表达阻断细胞的迁移与侵袭。Objective To explore the effect of nifuroxazide on proliferation,migration,and invasion of thyroid papillary carcinoma cells.Methods BCPAP and TPC-1cell lines treated with different concentration(0,1.25,2.5,5,10,20μmol/L)of nifuroxazide,respectively.Cell viability and proliferation of BCPAP and TPC-1was evaluated by MTT and colony formation assay.Apoptosis analysis and cell nuclear changes were determined by staining with Hoechst 33258 and visualized by a fluorescence microscope after treatment with nifuroxazide.Western blot analysis was used to evaluate protein expressions of apoptosis and invasion of BCPAP cells treated(48h)with nifuroxazide.Transwell assay was conducted to evaluate ability of cell migration and invasion.Results After being treated with nifuroxazide(0,1.25,2.5μmol/L and 0,1.25μmol/L)for 24,48,72 hrespectively,decreased proliferations of BCPAP and TPC-1cell lines were not obvious(P >0.05).However,treated BCPAP and TPC-1cells with higher concentration respectively(5,10,20μmol/L and 5,10μmol/L)of nifuroxazide for 24,48,72 h,the inhibitory effects were significantly obvious(P <0.05),and the inhibitory effects were increased in a concentration-and time-dependent manner.The inhibition in proliferation of TPC-1cell with nifuroxazide(2.5,5μmol/L)took effect from 72 hand 48h(P<0.05),respectively.Clone formations of BCPAP and TPC-1cells were significantly inhibited after being exposed to nifuroxazide(2.5,5μmol/L)for 10d(P<0.05).Hoechst33258 staining assay showed that nifuroxazide(10μmol/L)treatment resulted in cell shrinking,nuclear fragmentation and formation of condensed nuclei with bright-blue fluorescence.After 48 h,the percentage of apoptotic cells of BCPAP and TPC-1significantly increased respectively as the concentration of nifuroxazide with 10μmol/L(P<0.005).Pro-apoptotic protein CC-3and Bax expression levels increased significantly(P<0.05),and the expression of anti-apoptotic protein Bcl-2decreased significantly(P<0.05)in BCPAP cells after nifuroxazidetreatment(10μmol/L)for 48 h.The pe

关 键 词：甲状腺乳头状癌 硝呋齐特 细胞增殖 细胞迁移 细胞侵袭 

分 类 号：R736.1[医药卫生—肿瘤]