昆明医科大学学报
昆明醫科大學學報
곤명의과대학학보
Journal of Kunming Medical University
2014年
1期
5-7,20
,共4页
胡月新%徐天勇%张钰雯%赵瑜%田长富
鬍月新%徐天勇%張鈺雯%趙瑜%田長富
호월신%서천용%장옥문%조유%전장부
抑制%蛋氨酸裂解酶%协同%顺铂%肺腺癌细胞
抑製%蛋氨痠裂解酶%協同%順鉑%肺腺癌細胞
억제%단안산렬해매%협동%순박%폐선암세포
Inhibition%Methionine lyase%Synergy%Cisplatin%Lung adenocarcinoma cell
目的:纯化重组蛋氨酸酶并探讨其联合化疗药物顺铂对人肺腺癌GLC细胞生长的协同抑制效应.方法对原核表达体系pGEX-4T-1-Met/Dh5a菌液破碎后上清过纯化柱(GST)纯化后,采用MTT比色法测定重组蛋氨酸酶与顺铂联合应用对GLC细胞增殖的抑制作用,采用金正均法计算q值判断两药合用是否具有协同作用.结果重组蛋氨酸裂解酶浓度为0.22 mg/mL,纯度达95%,活性为0.568 IU/mg.给药72 h后单用顺铂对GLC细胞抑制率为24.80%,单用蛋氨酸酶抑制率为27.49%,两药合用抑制率为66.80%.=1.47>1.15,表现出明显协同作用.结论蛋氨酸酶及顺铂均有抑制GLC增殖作用,并且两者联合使用存在协同作用.
目的:純化重組蛋氨痠酶併探討其聯閤化療藥物順鉑對人肺腺癌GLC細胞生長的協同抑製效應.方法對原覈錶達體繫pGEX-4T-1-Met/Dh5a菌液破碎後上清過純化柱(GST)純化後,採用MTT比色法測定重組蛋氨痠酶與順鉑聯閤應用對GLC細胞增殖的抑製作用,採用金正均法計算q值判斷兩藥閤用是否具有協同作用.結果重組蛋氨痠裂解酶濃度為0.22 mg/mL,純度達95%,活性為0.568 IU/mg.給藥72 h後單用順鉑對GLC細胞抑製率為24.80%,單用蛋氨痠酶抑製率為27.49%,兩藥閤用抑製率為66.80%.=1.47>1.15,錶現齣明顯協同作用.結論蛋氨痠酶及順鉑均有抑製GLC增殖作用,併且兩者聯閤使用存在協同作用.
목적:순화중조단안산매병탐토기연합화료약물순박대인폐선암GLC세포생장적협동억제효응.방법대원핵표체체계pGEX-4T-1-Met/Dh5a균액파쇄후상청과순화주(GST)순화후,채용MTT비색법측정중조단안산매여순박연합응용대GLC세포증식적억제작용,채용금정균법계산q치판단량약합용시부구유협동작용.결과중조단안산렬해매농도위0.22 mg/mL,순도체95%,활성위0.568 IU/mg.급약72 h후단용순박대GLC세포억제솔위24.80%,단용단안산매억제솔위27.49%,량약합용억제솔위66.80%.=1.47>1.15,표현출명현협동작용.결론단안산매급순박균유억제GLC증식작용,병차량자연합사용존재협동작용.
Objective To purify recombinant methioninase and investigate the synergetic inhibitory effect of methioninase and cisplatin on the proliferation of human lung adenocarcinoma cell line GLC. Methods Recombinant methioninase was purified with GST-column from supernatant after ultrasonic disruption of cultured Escherichia coli in the prokaryotic expression system pGEX-4T-1-Met/Dh5a. MTT assay was used to determine the inhibition rate of methioninase in combined with cisplatin on cell proliferation,and their synergistic effect was evaluated by using the q value judge method. Results The concentration of recombinant methionine cleaving enzyme was 0.22 mg/mL, the purity was 95%, and the activity was 0.568 IU/mg. After 72 h culturing, the inhibition rate of cisplatin methionine was 24.80%and 27.49%respectively,while the inhibition rate of the combined drugs was 66.80% ( =1.47>1.15) which showed a significant synergistic effect. Conclusion Both methioninase and cisplatin have the inhibition effect,and the combined drugs display a significant synergistic effect on the proliferation of GLC.