摘要
目的观察雷帕霉素对人甲状腺乳头状癌细胞株K1增殖、迁移能力的影响,并探讨其可能机制。方法配制雷帕霉素和3-甲基腺嘌呤(3-MA)溶液,选择雷帕霉素作用浓度(对K1细胞的半抑制浓度)10μmol/L、3-MA作用浓度1 mmol/L进行后续实验。将K1细胞分为对照组、雷帕霉素组、雷帕霉素+3-MA组。对照组不做任何处理,加入2 mL培养基;雷帕霉素组加入100μmol/L雷帕霉素0.2 ml及培养基1.8 mL;雷帕霉素+3-MA组先加入100μmol/L雷帕霉素0.2 ml及培养基1.79 mL,1 h后加入1 mmol/L 3-MA溶液10 uL。培养24 h后,采用MTT法检测各组细胞的增殖能力,Transwell法检测各组细胞的迁移能力,Western blotting法检测各组细胞中的自噬相关蛋白LC3、p62。结果培养48、72、96 h后,雷帕霉素组细胞增殖能力低于对照组,雷帕霉素+3-MA组细胞增殖能力高于雷帕霉素组(P均<0.01)。雷帕霉素组穿膜细胞数少于对照组,雷帕霉素+3-MA组穿膜细胞数多于雷帕霉素组(P均<0.01)。雷帕霉素组LC3蛋白相对表达量高于对照组,p62蛋白相对表达量低于对照组(P均<0.01);雷帕霉素+3MA组LC3蛋白相对表达量低于雷帕霉素组,p62蛋白相对表达量高于雷帕霉素组(P均<0.01)。结论雷帕霉素可抑制甲状腺乳头状癌细胞的增殖、迁移能力,其机制可能与诱发自噬有关。
Objective To observe the effects of rapamycin on the proliferation and migration of human papillary thyroid carcinoma cell line K1 and to explore its possible mechanism. Methods We formulated rapamycin and 3-methyladenine（ 3-MA） solution,and selected 10 umol/L rapamycin（ semi inhibitory concentration of K1 cells） and 1 mmol/L 3-MA for the follow-up experiment. K1 cells were divided into the control group,rapamycin group,and rapamycin ＋ 3-MA group. The cells in the control group were only added with culture medium 2 mL,the cells in the rapamycin group were added with 100 mol/L rapamycin 0. 2 mL and culture medium 1. 8 mL,and the cells in the rapamycin ＋ 3-MA group were first added with rapamycin 0. 2 mL and culture medium 1. 79 mL,and an hour later,were added with 1 mmol/L 3-MA solution 10 u L. After 24-hour culture,the proliferation ability of each group was detected by MTT,the migration ability of each group was detected by Transwell,and the autophagy-related protein LC3 and p62 in each group was detected by Western blotting. Results At 48,72,96 h after culture,the cell proliferation of the rapamycin group was lower than that of the control group; the proliferation ability of rapamycin ＋ 3-MA group was higher than that of rapamycin group（ all P〈0. 01）. The number of transmembrane cells in the rapamycin group was less than that in the control group; the number of transmembrane cells in the rapamycin ＋ 3-MA group was more than that in the rapamycin group（ all P〈0. 01）. The relative expression of LC3 protein in the rapamycin group was higher than that in the control group,and the relative expression of p62 protein was lower than that of the control group（ both P〈0. 01）,and the relative expression of LC3 protein in the rapamycin ＋ 3 MA group was lower than that of the rapamycin group,and the relative expression of p62 protein was higher than that of the rapamycin group（ both P〈0. 01）. Conclusion Rapamycin can inhibit the proliferation and migration of papillary thyroid carcinoma cells,which may be related to the induction of autophagy.
作者
赵永魁
王晓涛
陈建立
王长友
张国志
ZHAO Yongkui;WANG Xiaotao;CHEN Jianli;WANG Changyou;ZHANG Guozhi(The Affiliated Hospital of North China University of Science and Technology,Tangshan 063000,Chin)
出处
《山东医药》
CAS
2018年第23期9-12,共4页
Shandong Medical Journal
基金
河北省卫计委科技成果推广项目(20150523)
关键词
甲状腺乳头状癌
雷帕霉素
细胞增殖
细胞迁移
自噬
微管相关蛋白轻链
P62
papillary thyroid carcinoma
rapamycin
cell proliferation
cell migration
autophagy
microtubule associated protein light chain
p62
