硼替佐米对人甲状腺癌细胞凋亡侵袭及AMPK/mTOR/4EBP1通路的影响  

Effects of Bortezomib on Apoptosis Invasion and AMPK/mTOR/4EBP1 Pathway in Human Thyroid Carcinoma Cells

作　　者：贺椿媛[1] 梁金环[1] 王翠 徐倩[1] HE Chunyuan;LIANG Jinhuan;WANG Cui(Department of Basic Medicine, Cangzhou Medical College, Hebei Cangzhou 061000, China)

机构地区：[1]沧州医学高等专科学校基础医学部生物化学教研室,河北沧州061000

出　　处：《河北医学》2022年第4期544-550,共7页Hebei Medicine

基　　金：河北省沧州市重点研发计划指导项目,(编号:172302090)。

摘　　要：目的:探究硼替佐米对人甲状腺癌(TC)细胞凋亡、侵袭的影响及可能的作用机制。方法:体外培养人甲状腺乳头状癌B-CPAP细胞,MTT法筛选硼替佐米的实验浓度及时间;取指数增长期的甲状腺癌B-CPAP细胞,分为对照组、AICAR组(AMPK激活剂2mmoL/L)、硼替佐米(50nmoL/L)组、硼替佐米+Compound C组(硼替佐米50nmoL/L+AMPK抑制剂Compound C 5μmoL/L);流式细胞仪检测各组B-CPAP细胞凋亡率;Transwell实验检测细胞侵袭能力;Western Blot检测细胞凋亡、侵袭以及AMPK/mTOR/4EBP1通路相关蛋白的表达[B细胞淋巴瘤/白血病-2基因(Bcl-2)、半胱氨酸天冬氨酸蛋白酶-3(Caspases-3)、E-钙黏连蛋白(E-cadherin)、β-连接素(β-catenin)、腺苷酸活化蛋白激酶(AMPK)、哺乳动物雷帕霉素靶蛋白(mTOR)、真核翻译起始因子4E结合蛋白1(4E-BP1)及其磷酸化蛋白]。结果:不同浓度的硼替佐米能不同程度的抑制B-CPAP细胞的生长,与对照组相比,AICAR组和硼替佐米组B-CPAP细胞凋亡率、Caspases-3、E-cadherin、β-catenin的表达、p-AMPK/AMPK的比值显著升高(P<0.05),进入Transwell小室下层的数量、Bcl-2的表达、p-mTOR/mTOR、p-4EBP1/4EBP1的比值显著降低(P<0.05),且AICAR组与硼替佐米组之间无明显统计学差异(P>0.05);使用AMPK抑制剂Compound C可升高p-mTOR/mTOR、p-4EBP1/4EBP1的比值,明显减弱硼替佐米对甲状腺癌B-CPAP细胞侵袭的抑制作用及促凋亡作用。结论:硼替佐米可抑制人甲状腺癌细胞增殖、侵袭,并诱导细胞凋亡,其作用机制可能与激活AMPK,抑制mTOR和4EBP1的活化有关。Objective:To explore the effects of bortezomib on apoptosis and invasion of human thyroid carcinoma(TC)cells and its possible mechanism.Methods:Human papillary thyroid carcinoma B-CPAP cells were cultured in vitro,and the experimental concentration and time of bortezomib were screened with MTT;thyroid carcinoma B-CPAP cells at the exponential growth stage were divided into control group,AICAR group(AMPK activator 2 mmol/L),bortezomib(50nmol/L)group,bortezomib+Compound C group(bortezomib 50nmol/L+AMPK inhibitor Compound C 5μmol/L);flow cytometry to detect the apoptosis rate of B-CPAP cells in each group;Transwell test to detect cell invasion ability;Western Blot to detect apoptosis,invasion and the expression of AMPK/mTOR/4EBP1 pathway-related proteins[B-cell lymphoma/leukemia-2 gene(Bcl-2),cysteine aspartic protease-3(Caspases-3),E-cadherin,β-catenin,adenosine monophosphate activated protein kinase(AMPK),mammalian target of rapamycin(mTOR),eukaryotic translation initiation factor 4E binding protein 1(4E-BP1)and its phosphorylated protein].Results:Different concentrations of bortezomib inhibited the growth of B-CPAP cells to varying degrees.Compared with the control group,the apoptosis rate of B-CPAP cells,the expression of Caspases-3,E-cadherin,β-catenin,and the ratio of p-AMPK/AMPK were significantly higher in the AICAR and bortezomib groups(P<0.05),and the entry Transwell lower layer,expression of Bcl-2,p-mTOR/mTOR,and p-4EBP1/4EBP1 ratio were significantly lower(P<0.05),and there was no statistically significant difference between the AICAR and bortezomib groups(P>0.05);the use of AMPK inhibitor Compound C elevated the ratios of p-mTOR/mTOR,p-4EBP1/4EBP1,and significantly reduced the inhibitory effect and the pro-apoptotic effect of bortezomib on the invasion of thyroid carcinoma B-CPAP cells.Conclusion:Bortezomib inhibits the proliferation and invasion of human thyroid carcinoma cells,and induce apoptosis.Its mechanism may be related to the activation of AMPK and inhibition of mTOR and 4EBP1 activation.

关 键 词：硼替佐米 甲状腺癌 凋亡 侵袭 

分 类 号：R73[医药卫生—肿瘤]