国际生物制品学杂志
國際生物製品學雜誌
국제생물제품학잡지
INTERNATIONAL JOURNAL OF BIOLOGICALS
2009年
4期
175-179
,共5页
白细胞介素类%抗肿瘤药%细胞凋亡%半胱氨酸天冬氨酸蛋白酶3
白細胞介素類%抗腫瘤藥%細胞凋亡%半胱氨痠天鼕氨痠蛋白酶3
백세포개소류%항종류약%세포조망%반광안산천동안산단백매3
Interleukins%Antineoplastic agent%Apoptosis%Caspases 3
目的 构建重组人白细胞介素24(recombinant human interleukin 24,rhIL-24)表达载体并在大肠杆菌中表达,体外评估rhIL-24的生物学活性.方法 用基因工程方法构建rhIL-24表达载体并在大肠杆菌巾表达rhIL-24.表达的重组蛋白经层析法纯化后,分别用噻唑蓝比色法和蛋白印迹法检测rhIL-24对肿瘤细胞生长的影响和对内源性IL-24的诱导,用实时PCR检测不同肿瘤细胞与rhIL-24共孵育后细胞内的胱天蛋白酶3(caspase 3)mRNA变化.结果 rhIL-24能在大肠杆菌中高效表达,并能明显抑制多种肿瘤细胞生长.rhIL-24能诱导正常MRC.5细胞和多种肿瘤细胞产生内源性IL-24,并能改变肿瘤细胞的胱天蛋白酶3水平.结论 rhIL-24能在大肠杆菌中高效表达,且具有生物学活性.
目的 構建重組人白細胞介素24(recombinant human interleukin 24,rhIL-24)錶達載體併在大腸桿菌中錶達,體外評估rhIL-24的生物學活性.方法 用基因工程方法構建rhIL-24錶達載體併在大腸桿菌巾錶達rhIL-24.錶達的重組蛋白經層析法純化後,分彆用噻唑藍比色法和蛋白印跡法檢測rhIL-24對腫瘤細胞生長的影響和對內源性IL-24的誘導,用實時PCR檢測不同腫瘤細胞與rhIL-24共孵育後細胞內的胱天蛋白酶3(caspase 3)mRNA變化.結果 rhIL-24能在大腸桿菌中高效錶達,併能明顯抑製多種腫瘤細胞生長.rhIL-24能誘導正常MRC.5細胞和多種腫瘤細胞產生內源性IL-24,併能改變腫瘤細胞的胱天蛋白酶3水平.結論 rhIL-24能在大腸桿菌中高效錶達,且具有生物學活性.
목적 구건중조인백세포개소24(recombinant human interleukin 24,rhIL-24)표체재체병재대장간균중표체,체외평고rhIL-24적생물학활성.방법 용기인공정방법구건rhIL-24표체재체병재대장간균건표체rhIL-24.표체적중조단백경층석법순화후,분별용새서람비색법화단백인적법검측rhIL-24대종류세포생장적영향화대내원성IL-24적유도,용실시PCR검측불동종류세포여rhIL-24공부육후세포내적광천단백매3(caspase 3)mRNA변화.결과 rhIL-24능재대장간균중고효표체,병능명현억제다충종류세포생장.rhIL-24능유도정상MRC.5세포화다충종류세포산생내원성IL-24,병능개변종류세포적광천단백매3수평.결론 rhIL-24능재대장간균중고효표체,차구유생물학활성.
Objective To construct an expression vector of recombinant human interleukin-24 (rhIL-24)and express it in E coli,and evaluate the biological activity in vitro.Methods An expression vector of rhIL-24 was constructed by genetic engineering and rhIL-24 Was expressed in E coli.After purification of rhIL-24 by chromatography,MTT assay was used to determine the effect of rhIL-24 on the growth of tumor cells and the Western blotting Was used to determine the induction of endogenous IL-24 by rhIL-24.rhIL-24 Was coincubated with different tumor cells.and real-time PCR was used to determine the change of easpase 3 mRNA.Results rhIL-24 was highly expressed in E coli,and the growth of tumor cells was inhibited significantly by rhIL-24.rhIL-24 could induce normal MRC-5 cells and tumor cells to produce endogenous IL-24,and change caspase 3 level of tumor cells.Conclusion rhIL-24 is highly expressed in E.coli and has biological activity.
