中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2009年
46期
3285-3288
,共4页
任明华%倪少滨%陈起引%徐万海%王长林%付宜鸣%焦治兴%麻立%赵忠山%刘兴汉
任明華%倪少濱%陳起引%徐萬海%王長林%付宜鳴%焦治興%痳立%趙忠山%劉興漢
임명화%예소빈%진기인%서만해%왕장림%부의명%초치흥%마립%조충산%류흥한
内皮抑素类%膀胱肿瘤%蛋白表达%抗肿瘤
內皮抑素類%膀胱腫瘤%蛋白錶達%抗腫瘤
내피억소류%방광종류%단백표체%항종류
Endostatins%Bladder neoplasms%Protein expression%Anti-tumor
目的 摸索重组人内皮抑素(rhES)蛋白的最佳表达条件.探索内皮抑素(Es)对膀胱癌实体肿瘤及细胞系的抑制作用.方法 分别从不同异丙基-β-D硫代半乳糖苷(IPTG)浓度、加IPTG 时机、表达时间等几个方面对 rhES 的蛋白表达条件进行摸索和优化.活性检测:(1)动物实验:观察内皮抑素对裸鼠皮下移植瘤的生长情况的影响.(2)细胞实验:MTT法、肿瘤细胞生长曲线观察内皮抑素对T-24膀胱癌细胞系的作用.结果 rhES 在大肠杆菌中的表达量占总菌体蛋白的58.65%,纯化后的蛋白纯度为96.22%.动物实验:rhES的抑瘤率为66.8%.细胞实验:MTT实验中rhES的半数抑制浓度IC_(50)=22μg/ml;肿瘤细胞生长曲线表明第7天时rhES组的细胞数比对照组减少了58.75%.结论 本实验所优化的最佳表达条件可获得高表达、高活性的rh Es蛋白,Es不但对裸鼠接种的膀胱癌有抑制作用,对膀胱癌细胞系也有抑制作用.
目的 摸索重組人內皮抑素(rhES)蛋白的最佳錶達條件.探索內皮抑素(Es)對膀胱癌實體腫瘤及細胞繫的抑製作用.方法 分彆從不同異丙基-β-D硫代半乳糖苷(IPTG)濃度、加IPTG 時機、錶達時間等幾箇方麵對 rhES 的蛋白錶達條件進行摸索和優化.活性檢測:(1)動物實驗:觀察內皮抑素對裸鼠皮下移植瘤的生長情況的影響.(2)細胞實驗:MTT法、腫瘤細胞生長麯線觀察內皮抑素對T-24膀胱癌細胞繫的作用.結果 rhES 在大腸桿菌中的錶達量佔總菌體蛋白的58.65%,純化後的蛋白純度為96.22%.動物實驗:rhES的抑瘤率為66.8%.細胞實驗:MTT實驗中rhES的半數抑製濃度IC_(50)=22μg/ml;腫瘤細胞生長麯線錶明第7天時rhES組的細胞數比對照組減少瞭58.75%.結論 本實驗所優化的最佳錶達條件可穫得高錶達、高活性的rh Es蛋白,Es不但對裸鼠接種的膀胱癌有抑製作用,對膀胱癌細胞繫也有抑製作用.
목적 모색중조인내피억소(rhES)단백적최가표체조건.탐색내피억소(Es)대방광암실체종류급세포계적억제작용.방법 분별종불동이병기-β-D류대반유당감(IPTG)농도、가IPTG 시궤、표체시간등궤개방면대 rhES 적단백표체조건진행모색화우화.활성검측:(1)동물실험:관찰내피억소대라서피하이식류적생장정황적영향.(2)세포실험:MTT법、종류세포생장곡선관찰내피억소대T-24방광암세포계적작용.결과 rhES 재대장간균중적표체량점총균체단백적58.65%,순화후적단백순도위96.22%.동물실험:rhES적억류솔위66.8%.세포실험:MTT실험중rhES적반수억제농도IC_(50)=22μg/ml;종류세포생장곡선표명제7천시rhES조적세포수비대조조감소료58.75%.결론 본실험소우화적최가표체조건가획득고표체、고활성적rh Es단백,Es불단대라서접충적방광암유억제작용,대방광암세포계야유억제작용.
Objective To explore the optimal method for protein expression in rhES (recombinant human endostatin ) and study the anti-tumor activities of rhES in solid tumor and established cell line.Methods Different IPTG concentrations.the timing of adding IPTG into the culture medium and the different time of expression were employed to explore the optimized conditions of protein expression.Activity examination:(1) animal experiment:nude mice bearing subcutaneous cancer in treated group and controlled group were observed.(2) cellular experiment:the inhibitory effect of rhES in T-24 established cell line were observed by MTT assay and cancer cell growth curve.ResulIs The expression of rhES protein was 58.65%.Of all the E.coli body proteins.the protein purity after purification was 96.22%.Activity examination indicated that rhES could inhibit the growth of solid tumor and the established cell line.In animal experiment.the tumor inhibition rate Was 66.8%.Cell experiment:the 50% inhibitory concentration (IC_(50)) was 22 μg/ml.The cell population decreased 58.75% than the control group at Day 7 in the tumor cell growth curve.Conclusion A high expression and activity of rhES protein can be obtained by the optimized expression conditions.rhES can inhibit the cellular growth in both solid tumor and established cell line of bladder cancer.
