中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2010年
2期
123-127
,共5页
王伟%陈平%李军利%裴艳芳%双庆翠%刘彩虹%蔡珊%刘绍坤%诸兰艳%周锐
王偉%陳平%李軍利%裴豔芳%雙慶翠%劉綵虹%蔡珊%劉紹坤%諸蘭豔%週銳
왕위%진평%리군리%배염방%쌍경취%류채홍%채산%류소곤%제란염%주예
肺肿瘤%顺铂%蛋白激酶类%Tumstatin
肺腫瘤%順鉑%蛋白激酶類%Tumstatin
폐종류%순박%단백격매류%Tumstatin
Lung neoplasms%Cisplatin%Protein kinases%Tumstatin
目的 观察Tumstatin185～191对肺腺癌细胞增生与凋亡的影响,探讨其与蛋白激酶B(Akt)及细胞外调节蛋白激酶(ERK)活性的关系.方法 以人肺腺癌细胞株A549为研究对象,分别给予不同浓度的Tumstatin185～191、顺铂及Tumstatin185～191联合顺铂进行干预,以四唑盐比色法测定A549细胞的增殖情况;流式细胞仪检测分析A549细胞凋亡的变化;Western blot检测细胞内磷酸化的Akt(p-Akt)及磷酸化的ERK(p-ERK)蛋白表达水平.结果 Tumstatin185～191对A549细胞的半数抑制浓度(IC_(50))为73.7μmol/L,顺铂对A549细胞的IC_(50)为5.2μmol/L,当顺铂与20μmol/L或40μmol/L的Tumstatin185～191联合应用时,顺铂对A549细胞的IC_(50)分别下降为3.5 μmol/L和1.4μmol/L;两药联合使用时,A549细胞的早期凋亡率为(19.34±0.97)%,较顺铂[(12.5±2.1)%]和Tumstatin185～191单用[(9.6±1.6)%]的早期凋亡率显著增加(F=5.74,P<0.01);Western blot检测显示p-Akt及p-ERK在未经干预的A549细胞内均呈高表达状态,Tumstatin185～191可显著抑制p-Akt及p-ERK的表达,顺铂则无明显影响;Tumstatin185～191与顺铂联合使用并不增加Tumstatin185～191对p-Akt及p-ERK的抑制效应.结论 Tumstatin185～191单药或联合顺铂对于A549细胞均有显著抑制作用,Tumstatin185～191能够增加A549细胞对顺铂的敏感性;Tumstatin185～191可能通过下调A549细胞内p-Akt及p-ERK水平发挥其抑制细胞生长、促进细胞凋亡的作用.
目的 觀察Tumstatin185～191對肺腺癌細胞增生與凋亡的影響,探討其與蛋白激酶B(Akt)及細胞外調節蛋白激酶(ERK)活性的關繫.方法 以人肺腺癌細胞株A549為研究對象,分彆給予不同濃度的Tumstatin185～191、順鉑及Tumstatin185～191聯閤順鉑進行榦預,以四唑鹽比色法測定A549細胞的增殖情況;流式細胞儀檢測分析A549細胞凋亡的變化;Western blot檢測細胞內燐痠化的Akt(p-Akt)及燐痠化的ERK(p-ERK)蛋白錶達水平.結果 Tumstatin185～191對A549細胞的半數抑製濃度(IC_(50))為73.7μmol/L,順鉑對A549細胞的IC_(50)為5.2μmol/L,噹順鉑與20μmol/L或40μmol/L的Tumstatin185～191聯閤應用時,順鉑對A549細胞的IC_(50)分彆下降為3.5 μmol/L和1.4μmol/L;兩藥聯閤使用時,A549細胞的早期凋亡率為(19.34±0.97)%,較順鉑[(12.5±2.1)%]和Tumstatin185～191單用[(9.6±1.6)%]的早期凋亡率顯著增加(F=5.74,P<0.01);Western blot檢測顯示p-Akt及p-ERK在未經榦預的A549細胞內均呈高錶達狀態,Tumstatin185～191可顯著抑製p-Akt及p-ERK的錶達,順鉑則無明顯影響;Tumstatin185～191與順鉑聯閤使用併不增加Tumstatin185～191對p-Akt及p-ERK的抑製效應.結論 Tumstatin185～191單藥或聯閤順鉑對于A549細胞均有顯著抑製作用,Tumstatin185～191能夠增加A549細胞對順鉑的敏感性;Tumstatin185～191可能通過下調A549細胞內p-Akt及p-ERK水平髮揮其抑製細胞生長、促進細胞凋亡的作用.
목적 관찰Tumstatin185～191대폐선암세포증생여조망적영향,탐토기여단백격매B(Akt)급세포외조절단백격매(ERK)활성적관계.방법 이인폐선암세포주A549위연구대상,분별급여불동농도적Tumstatin185～191、순박급Tumstatin185～191연합순박진행간예,이사서염비색법측정A549세포적증식정황;류식세포의검측분석A549세포조망적변화;Western blot검측세포내린산화적Akt(p-Akt)급린산화적ERK(p-ERK)단백표체수평.결과 Tumstatin185～191대A549세포적반수억제농도(IC_(50))위73.7μmol/L,순박대A549세포적IC_(50)위5.2μmol/L,당순박여20μmol/L혹40μmol/L적Tumstatin185～191연합응용시,순박대A549세포적IC_(50)분별하강위3.5 μmol/L화1.4μmol/L;량약연합사용시,A549세포적조기조망솔위(19.34±0.97)%,교순박[(12.5±2.1)%]화Tumstatin185～191단용[(9.6±1.6)%]적조기조망솔현저증가(F=5.74,P<0.01);Western blot검측현시p-Akt급p-ERK재미경간예적A549세포내균정고표체상태,Tumstatin185～191가현저억제p-Akt급p-ERK적표체,순박칙무명현영향;Tumstatin185～191여순박연합사용병불증가Tumstatin185～191대p-Akt급p-ERK적억제효응.결론 Tumstatin185～191단약혹연합순박대우A549세포균유현저억제작용,Tumstatin185～191능구증가A549세포대순박적민감성;Tumstatin185～191가능통과하조A549세포내p-Akt급p-ERK수평발휘기억제세포생장、촉진세포조망적작용.
Objective To investigate the antitumor effects of tumstatin 185-191 as a single agent or combination with cisplatin(DDP) on non-small lung cancer(NSCLC) cell lines A549. In addition, the changes of the protein kinase B(Akt) and extracelluarregulated protein kinase(ERK) in cultured NSCLC cells treated by tumstatin 185-191 and cisplatin were evaluated. Methods A549 cells were treated with tumstatin185 -191 and cisplatin. Cell viability was assessed using the modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell apoptosis was measured by flow cytometry. The activation of Akt and Erk were evaluated by Western blotting. Results Tumstatin185 - 191 inhibited the proliferation of A549 and the IC_(50) values of tumstatin185 - 191 was 73. 7 μmol/L. After cotreatment with 20 μmol/L tumstatin185 - 191, IC_(50) values of cisplatin in A549 cells reduced from 5.2 μmol/L to 3. 5 μmol/L, while 40 μmol/L tumstatin185 - 191 reduced from 5.2 μmol/L to 1.4 μmol/L. The early apoptesis rate was (19.34± 0.97)% in the cotreatment group, (12.5±2.1)% in cisplatin group and (9.6±1.6)% in tumstatin185 -191 group (F = 5.74, P <0.01). The levels of phospho-Akt (p-Akt) and phospho-ERK (p-ERK) in the A549 cells were remarkably lower after being treated with tumstatin 185-191, while tumstatin 185 - 191 treatment whether alone, or in combination with cisplatin, had the similar effects on the protein levels of p-Akt and p-ERK in A549 cells. Conclusion Our data suggest that tumstatin185 - 191 might enhance the sensitivity of A549 cells to cisplatin. The effects of promoting apoptosis and downregulation of proliferation induced by tumstatin185 - 191 may be mediated through inactivation of the Akt and ERK pathways.
