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1. (1.重庆医科大学实验动物中心,重庆,400016
2. 重庆医科大学附属第一医院分子肿瘤及表观遗传学实验室,重庆,400016
3. 3.泸州医学院药物与功能性食品研究中心,泸州,646000
纸质出版日期:2012,
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潘玥, 向廷秀, 章海林, 等. 真核双基因表达载体pBud-TRAIL-IL-24的构建及其在宫颈癌细胞中的表达[J]. 重庆医科大学学报, 2012,37(4):302-306.
PAN Yue, XIANG Tingxiu, ZHANG Hailin, et al. Construction of eukaryotic co expression plasmid pBud-TRAIL-IL-24 and its expression in cervical cancer cells[J]. Journal of Chongqing Medical University, 2012,37(4):302-306.
潘玥, 向廷秀, 章海林, 等. 真核双基因表达载体pBud-TRAIL-IL-24的构建及其在宫颈癌细胞中的表达[J]. 重庆医科大学学报, 2012,37(4):302-306. DOI:
PAN Yue, XIANG Tingxiu, ZHANG Hailin, et al. Construction of eukaryotic co expression plasmid pBud-TRAIL-IL-24 and its expression in cervical cancer cells[J]. Journal of Chongqing Medical University, 2012,37(4):302-306. DOI:
目的:构建肿瘤坏死因子相关凋亡诱导配体(Tumor necrosis factor related apoptosis inducing ligand
TRAIL)基因和白细胞介素24(Interleutin-24
IL-24)基因的真核双基因表达载体,并观察其对宫颈癌细胞生长的影响情况。方法:采用分子克隆技术,将IL-24基因插入真核表达载体 pBud-TRAIL中,构建双表达质粒 pBud-TRAIL-IL-24,经酶切和测序鉴定后,转染宫颈癌HeLa细胞。荧光显微镜和免疫荧光双标技术检测TRAIL和IL-24在HeLa细胞中的表达。MTT法检测重组质粒对HeLa细胞生长状况的影响。结果:成功构建了真核双基因表达载体pBud-TRAIL-IL-24,并在宫颈癌细胞中表达;转染48 h后可见重组质粒对宫颈癌细胞生长有明显抑制作用(P<0.05)。结论:构建的双表达载体pBud-TRAIL-IL-24能在宫颈癌细胞中表达,并可有效抑制宫颈癌HeLa细胞增殖,为进一步研究TRAIL和IL-24联合应用于宫颈癌的基因治疗奠定了基础。
Objective:To construct the eukaryotic co expression plasmid encoding TNF related apoptosis inducing ligand(TRAIL)gene and interleukin 24(IL-24)gene and to study its inhibitory effect on cervical cancer cell lines.Methods:IL-24 gene was inserted into the expression vector of pBud-TRAIL to obtain co-expression plasmid pBud-TRAIL-IL-24 by molecular cloning technology.The constructed co-expression plasmid was identified by restriction endoenzyme digestion and nucleotides sequencing.The recombinant plasmid of pBud-TRAIL-IL-24 was transfected into HeLa cells.The expressions of TRAIL and IL-24 were detected by fluorescence microscopy and immunofluorescence double labeling staining.The proliferation of cervical cancer cells was assessed by MTT assay.Results:The eukaryotic co-expression plasmid pBud-TRAIL-IL-24 could be successfully constructed and be efficiently co-expressed in HeLa cells.The recombinant plasmid significantly inhibited the growth of cervical cancer cells(P<0.05).Conclusion:The plasmid pBud-TRAIL-IL-24 can be successfully constructed and efficiently co-expressed in HeLa cells
and it can effectively inhibit the proliferation of HeLa cells
which lays the foundation for further study of the joint application of TRAIL and IL-24 in cervical cancer gene therapy.
宫颈癌细胞双表达载体TRAIL基因IL-24基因
cervical cancerco-expression vectortumor necrosis factor related apoptosis inducing ligand gene(TRAIL-gene)interleutin-24(IL-24)
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