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人IL-24基因在CHO细胞中的表达及其抗肿瘤效应 被引量：11

Expression of human IL-24 gene in CHO cells and its anti-tumor effect in vitro
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摘要 目的构建人IL-24基因真核表达载体,在CHO细胞中进行稳定表达,并检测重组表达蛋白rhIL-24的抗肿瘤活性。方法将测序验证的人IL-24基因亚克隆至真核表达载体pcDNA3,构建重组真核表达载体pcDNA3-hIL-24,转染CHO细胞进行稳定表达,经RT-PCR鉴定后用MTT法、Ho-echst染色和流式细胞术检测CHO细胞表达的rhIL-24诱导A549人肺腺癌细胞凋亡的抗肿瘤效应,用ELISA检测其刺激免疫细胞分泌IL-6和IFN-γ的功能。结果经双酶切和PCR鉴定,重组真核表达载体构建正确,人IL-24在CHO细胞中获得稳定表达,且所表达的人IL-24具有较强的诱导A549人肺腺癌细胞凋亡及上调免疫细胞表达IL-6和IFN-γ的免疫刺激活性。结论人IL-24基因的稳定表达和抗肿瘤效应的实验研究,为进一步研究人IL-24抗肿瘤的分子机制及潜在的应用奠定了基础。 Aim To construct the eukaryotic expression vector of hIL-24 cDNA , and express it in CHO cells and detect its anti-tumor effect of recombinant hIL-24 protein. Methods Constructed peDNA3-hIL-24 was identified by endonueleases digestion ＆ PCR. The recombinant expression plasmids were transfeeted into CHO cells, human hIL-24 expressed in CHO cells was detected with RT-PCR. The apoptosis-indueing activities of recombinant protein hIL-24 was tested by MTr assay , Hoeehst＆ FCM assay, and the expression of IL- 6 and IFN-γ/from PBMC induced by rhIL-24 was tested by ELISA. Results The eukaryotie expression vector pcDNA3-hIL-24 was constructed correctly. Stable expression of human IL-24 in CHO cells was identified with RT-PCR. The apoptosis of A549 cells induced by hIL-24 was proved by Hoechst ＆ FCM assay, and the expression of IL-6 and IFN-γ/from PBMC induced by rhIL-24 was identified with ELISA. Conclusion The successful stable expression ＆ experimental study of apoptosis effect of human IL-24 gene lay the foundation for the further study of molecular mechanism of hIL-24 on anti-tumors and potential application.
作者 缪竞诚 陈雄艳 盛伟华 谢宇锋 杨吉成
机构地区 苏州大学医学院细胞和分子生物学教研室
出处 《中国药理学通报》 CAS CSCD 北大核心 2006年第5期567-570,共4页 Chinese Pharmacological Bulletin
基金 苏州大学医学发展基金资助项目(NoEE134517)
关键词 人白细胞介素-24 稳定转染 CHO细胞 凋亡 human IL-24 stable transfection CHO cell apoptosis
分类号 R-329.25 [医药卫生] R342.3 [医药卫生—基础医学]
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参考文献12

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  • 2Jiang H,Lin J J,Su Z Z,et al.Substraction hybridization identifies a novel melanoma-differentiation-associated gene,mda-7,modulated during human melanoma differentiation,growth and progression[J].Oncogene,1995,11(12):2477-86.
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  • 7陈雄艳,盛伟华,谢宇锋,缪竞诚,白艳艳,杨吉成.人IL-24基因的克隆及在COS-7细胞中的表达[J].中国免疫学杂志,2004,20(10):672-676. 被引量：12
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二级参考文献17

  • 1Kiang H, Ling JJ, Su ZZ , et al. Substraction hybridization identifies a novel melanoma- differentiation - associated gene, mda- 7, modulated during human melanoma differentiation, growth and progression [ J ]. Oncogene,1995, 11(12) :2477-2486.
  • 2Candell EG, Mumm JB, Poindexter N, et al. The protein product of the humor suppressor gene, melanoma differentiation - associated gene 7, exhibits immunostimulatory activity and isdesignated IL - 24 [J]. J Immunol, 2002,168(12) : 6041- 6046.
  • 3Madireddi MT, Dent P, Fisher PB, et al. Regulation of mda- 7 gene expression during human melanoma differentiation[J]. Oncogene, 2000, 19(10) : 1362-- 1368.
  • 4Ellerhorst JA, Prieto VG, Ekmekcioglu S, et al. Loss of MDA-7 expression with progression of melanoma[J]. J Clin Oncol, 2002, 20(4) : 1069-1074.
  • 5Lebedeva IV, Su ZZ, Chang Y , et al. The cancer growth suppressing gene mda- 7 induces apoptosis selectively in human melanoma calls[J]. Oncogene, 2002 , 21 (5) :708-718.
  • 6Kawabe S, Nishikawa T, Munshi A, et al. Adenovirusmediated mda - 7 gene expression radiosensitizes nonsmall cell lung cancer cells via TP53 - independent mechanisms[J]. Mol Ther, 2002 , 6(5) : 637-- 644.
  • 7Su Z, Lebedeva IV, Gopalkrishnan RV, et al. A combinatorial approach for selectively inducing programmed cell death in human pancreatic cancer cells[J]. Proc Natl Acad Sci USA, 2001, 98(18):10332--10337.
  • 8Fickenscher H,Hor S,Kupers H et al.The interleukin-10 family of cytokines [J].Trends Immunol,2002;23(2):89-96.
  • 9Jiang H,Lin J J,Su Z Z et al.Substraction hybridization identifies a novel melanoma-differentiation-accociated gene,mda-7,modulated during human melanoma differentiation,growth and progression [J].Oncogene,1995;11(12):2477-2486.
  • 10Lebedeva I V,Su Z Z,Chang Y et al.The cancer growth suppressing gene mda-7 induces apoptosis selectively in human melanoma cells [J].Oncogene,2002;21(5):708-718.

共引文献17

同被引文献134

引证文献11

二级引证文献24

中国药理学通报
2006年 第5期

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