CAJ | 학술논문

目的研究新型酪氨酸激酶抑制剂帕唑帕尼对甲状腺癌细胞株SW579和血管内皮细胞株HUVEC增殖的影响及作用机制.方法将SW579和HUVEC暴露于不同浓度(1、2、4、8和16 μmoL/L)的帕唑帕尼至72 h,无药物处理的细胞作为对照.采用细胞活度测定、图像分析技术及分裂细胞荧光免疫染色观察评价帕唑帕尼对细胞增殖的影响,应用流式细胞仪进行细胞周期时相分析.结果细胞活度测定显示,帕唑帕尼对SW579和HUVEC细胞的生长增殖均有明显的抑制作用,并呈浓度依赖性及时间依赖性,其半数抑制浓度(IC50)分别约为2和4μmol/L.图像分析显示,在IC50作用下,帕唑帕尼使两种细胞的细胞密度降低,分裂指数下降.帕唑帕尼作用48 h时,SW579细胞分裂指数(0.9％)明显低于其对照细胞(2.2％),差异有统计学意义(P＜0.01);HUVEC细胞分裂指数(1.9％)亦明显低于其对照细胞(4.0％),差异有统计学意义(P＜0.01).细胞周期时相分析表明,帕唑帕尼可使SW579和HUVEC发生细胞周期休止,分别休止于G1期和S期.帕唑帕尼作用24、48 h时,SW579在G1期的比例均明显高于其对照组细胞[(60.4±2.2)％比(49.2±1.6)％,(70.4±1.6)％比(58.6±3.3)％],差异有统计学意义(P＜0.01);HUVEC细胞在S期的比例均明显高于其对照组细胞[(47.3±2.7)％比(39.5±0.6)％,(43.5±1.6)％比(28.8±4.2)％],差异有统计学意义(P＜0.01).结论帕唑帕尼对甲状腺癌细胞和血管内皮细胞的增殖均有明显的抑制效应,其作用是藉细胞周期休止作用而实现,但在两种细胞细胞周期休止时相不同.
목적 연구신형락안산격매억제제파서파니대갑상선암세포주SW579화혈관내피세포주HUVEC증식적영향급작용궤제.방법 장SW579화HUVEC폭로우불동농도(1、2、4、8화16 μmoL/L)적파서파니지72 h,무약물처리적세포작위대조.채용세포활도측정、도상분석기술급분렬세포형광면역염색관찰평개파서파니대세포증식적영향,응용류식세포의진행세포주기시상분석.결과 세포활도측정현시,파서파니대SW579화HUVEC세포적생장증식균유명현적억제작용,병정농도의뢰성급시간의뢰성,기반수억제농도(IC50)분별약위2화4μmol/L.도상분석현시,재IC50작용하,파서파니사량충세포적세포밀도강저,분렬지수하강.파서파니작용48 h시,SW579세포분렬지수(0.9％)명현저우기대조세포(2.2％),차이유통계학의의(P＜0.01);HUVEC세포분렬지수(1.9％)역명현저우기대조세포(4.0％),차이유통계학의의(P＜0.01).세포주기시상분석표명,파서파니가사SW579화HUVEC발생세포주기휴지,분별휴지우G1기화S기.파서파니작용24、48 h시,SW579재G1기적비례균명현고우기대조조세포[(60.4±2.2)％비(49.2±1.6)％,(70.4±1.6)％비(58.6±3.3)％],차이유통계학의의(P＜0.01);HUVEC세포재S기적비례균명현고우기대조조세포[(47.3±2.7)％비(39.5±0.6)％,(43.5±1.6)％비(28.8±4.2)％],차이유통계학의의(P＜0.01).결론 파서파니대갑상선암세포화혈관내피세포적증식균유명현적억제효응,기작용시자세포주기휴지작용이실현,단재량충세포세포주기휴지시상불동.
Objective To investigate the in vitro effects of pazopanib,a tyrosine kinase inhibitor,on the proliferations of both thyroid cancer cells and vascular endothelial cells and underlying mechanisms.Methods The cultured human anaplastic thyroid carcinoma cell line SW579 and human umbilical vein endothelial cell line HUVEC were exposed to pazopanib at different concentrations (1,2,4,8 and 16 μmol/L) for up to 72 hours,and the pazopanib-untreated cells were as control.The effects of pazopanib on the cell proliferations were assessed by cell viability assay,cell imaging analysis and mitosis index measure with immunofluorecent staining of mitosisi cells.Cell cycle analysis was performed by flow cytometry.Results Cell viability assay indicated that pazopanib inhibited the proliferations of both SW579 and HUVEC,in time-and dose-dependent manners,with the estimated IC50 of 2 μmol/L and 4 μmol/L,respectively.Cell imaging analysis showed that pazopanib at IC50 inhibited significantly the growths of SW579 and HUVEC,with significant decreased in the cell densities.Immunofluorecent staining for mitosis cells demonstrated that the mitosis index (0.9％) of the pazopanib-treated SW579 at 48 hours was lower significantly than that (2.2％) of control cells (P ＜ 0.01),and also the mitosis index (1.9％) of pazopanib-treated HUVEC at 48 hours was lower significantly than that of control cells (4.0％) (P ＜ 0.01).Flow cytometry demonstrated that pazopanib induced SW579 cells to arrest at G1 phase.At 24 hours and 48 hours,the percentages of pazopanib-treated SW579 in G1 phase were higher significantly than that of control group cells [(60.4 ± 2.2) ％ vs (49.2 ± 1.6) ％,(70.4 ± 1.6) ％ vs (58.6 ± 3.3) ％] (P ＜ 0.01),while the percentages of pazopanib-treated HUVEC in S phase were higher significantly than that of control group cells [(47.3 ± 2.7) ％ vs (39.5 ±0.6)％,(43.5 ±1.6)％ vs (28.8 ±4.2)％] (P9＜0.01).Conclusions Pazopanib inhibits the proliferations of both SW579 and HUVEC by inducing cell cycle arrest at different phases,leading G1 block of SW579 and S phase arrest of HUVEC respectively.Pazopanib is indicated as a new tyrosine kinase inhibitor to be used for the treatment of thyroid cancer by targeting both tumor cells and vascular endothelial cells related to tumor angiogenesis.