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探讨克服肿瘤细胞多药耐药(MDR1)性的方法,提高化疗效果。本文采用多药耐药反义基因(MDR1-RSPS-ODN)逆转K562/ADM肿瘤细胞的MDR1,诱导肿瘤细胞凋亡,发现MDR1-ASPS-ODN诱导K562/ADM细胞株细胞产生大量DNA断片,FACS检测发现几乎全部MRD+K562/ADM细胞发生凋亡。其结果表明DMDR1-ASPS-ODN能有效、特异地抑制MDR1基因表达,逆转肿瘤细胞的MDR1,促进阿霉素诱导MDR+1K562/ADM细胞凋亡,为其临床应用提供理论依据 相似文献
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目的:探讨聚乙烯亚胺(polyethylene im ine,PEI)作为生存素(survivin)反义核酸(anti-sense oligodeoxynuc leotides,ASODN)的载体投递肝癌SMMC-7721细胞的条件。方法:凝胶阻滞实验筛选PEI与survivin ASODN形成静电复合物的最佳质量比;W ST-8法检测PEI对肝癌细胞的毒性;流式细胞仪及W ST-8法筛选荧光标记的PEI与ASODN在不同质量比时的瞬时转染效率及48 h转染效率;流式细胞仪及荧光显微镜分别检测荧光标记的ASODN以及PEI-ASODN复合物进入细胞的情况;W ST-8法检测血清对PEI转染效率的影响。结果:m(PEI)∶m(ASODN)为0.625∶1~2.5∶1时可以形成电中性状态的静电复合物;PEI终质量浓度≤4μg/mL对肝癌细胞的毒性较小;m(PEI)∶m(ASODN)为0.75∶1时转染效率最高;血清存在对PEI转染效率无明显影响。结论:PEI是一种低毒的ASODN投递载体,其终质量浓度≤4μg/mL可作为肝癌细胞的转染载体,血清条件下m(PEI)∶m(ASODN)为0.75∶1时,PEI携带sur... 相似文献
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Alternative splicing of Bcl-2-related genes: functional consequences and potential therapeutic applications 总被引:6,自引:0,他引:6
Apoptosis is a morphologically distinct form of cell death. It is executed and regulated by several groups of proteins. Bcl-2 family proteins are the main regulators of the apoptotic process acting either to inhibit or promote it. More than 20 members of the family have been identified so far and most have two or more isoforms. Alternative splicing is one of the major mechanisms providing proteomic complexity and functional diversification of the Bcl-2 family proteins. Pro- and anti-apoptotic Bcl-2 family members should function in harmony for the regulation of the apoptosis machinery, and their relative levels are critical for cell fate. Any mechanism breaking down this harmony by changing the relative levels of these antagonistic proteins could contribute to many diseases, including cancer and neurodegenerative disorders. Recent studies have shown that manipulation of the alternative splicing mechanisms could provide an opportunity to restore the proper balance of these regulator proteins. This review summarises current knowledge on the alternative splicing products of Bcl-2-related genes and modulation of splicing mechanisms as a potential therapeutic approach.Received 5 January 2004; received after revision 31 March 2004; accepted 6 April 2004 相似文献
4.
cDNA cloning and functional analysis of 4-coumarate --CoA: ligase (4CL) gene in Chinese white aspen 总被引:5,自引:0,他引:5
cDNA encoding 4-coumarate: CoA ligase (4CL) was isolated from the secondary developing xylem of Chinese white aspen (Populus tomentosa) by RT-PCR for the first time, which was 1619 bp in length. The coding sequence and putative amino acid sequence showed 97.53% and 97.00% identity to that of Pt4CL1 in quaking aspen (P. tremuloids), respectively. Molecular analysis indicated that 4CL was encoded by multiple genes in P.tomentosa, its mRNA was highly accumulated in xylem and the expression of 4CL revealed a biphasic pattern in one growing season, almost in phase with the expression of other related enzymes in lignin biosynthesis. Transgenic research showed that expression of antisense 4CL cDNA led to the decreasing of lignin content in transgenic tobaccos, among which the average reduction was 10.3% and the highest could be up to 18.9%. These data suggested that 4CL gene was a potential gene used in altering lignin biosynthesis by biotechnology for producing new materials of papermaking. 相似文献
5.
LUO Jia HUANG YanYan XIONG ShaoXiang LIU GuoQuan ZHAO Ruit 《科学通报(英文版)》2007,52(10):1311-1319
The specific interaction between sense and antisense peptides was studied by high-performance affinity chromatography (HPAC) and quartz crystal microbalance (QCM) biosensor. Fragment 1-14 of human interferon-β (hlFN-βwas chosen as sense peptide and its three antisense peptides (AS-IFN 1, AS-IFN 2, and AS-IFN 3) were designed according to the degeneracy of genetic codes. The affinity column was prepared with sense peptide as ligand and the affinity chromatographic behavior was evaluated. Glu-substituted antisense peptide (AS-IFN 3) showed the strongest binding to immobilized sense peptide at pH 7.5. A quartz crystal microbalance-flow injection analysis (QCM-FIA) system was introduced to investigate the recognition process in real-time. The equilibrium dissociation constants between sense peptide and AS-IFN 1, AS-IFN 2 and AS-IFN 3 measured 2.08×10^-4, 1.31×10^-4 and 2.22×10^-5 mol/L, respectively. The mechanism study indicated that the specific recognition between sense peptide and AS-IFN 3 was due to sequence-dependent and multi-modal affinity interaction. 相似文献
6.
To study the effect of the major histocompatibility complex class II (MHC II) transactivator (CIITA) antisense RNA on the
expression of the human leukemia (HLA) class II molecules, 5′ end cDNA sequence of CIITA gene was cloned, and antisense RNA
expression vector pcDNA-II was constructed. HeLa cells transfected with pcDNA-II and pcDNA3 were induced by IFN-γ for 3 d.
The expression of HLA class II molecules on HeLa/pcDNA-II cells was significantly decreased, while it has no effect on the
expression of HLA class I molecules. This result suggests that the CIITA antisense RNA can inhibit the expression of HLA class
II molecules in HeLa cells. It also implies a promising approach to generate immune tolerance in graft transplantation. 相似文献
7.
Cellular apoptosis susceptibility (CAS) gene plays important roles in mitosis, development and export of importin a from the nucleus, but its function in plant is unknown. In this study, a rice CAS ortholog (OsCAS), which encodes a predicted protein of 983 amino acids with 62% similarity to human CAS, was identified. DNA gel blot analysis revealed a single copy of OsCAS in the rice genome. A 973 bp fragment at the 3' end of OsCAS cDNA was cloned from rice cDNA library and transferred into rice in the antisense direction under the control of CaMV 35S promoter via Agrobacterium-mediated transformation method, 105 transgenic lines were obtained. Expression of OsCAS was suppressed in the antisense transgenic lines as revealed by semi-quantitative RT-PCR. The antisense transgenic lines showed dwarf phenotypes. The results indicated that OsCAS was involved in culm development of rice. 相似文献
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SPIO标记的反义探针磁学性能检测 总被引:1,自引:0,他引:1
采用化学交联法制备超顺磁性氧化铁(SPIO)标记的c-erbB2癌基因反义脱氧寡核苷酸探针,经过1.5T磁共振仪测定,其T2弛豫率为0.156×106mol-1·S-1;振动样品磁强计测定出探针的磁化强度为69.423 8 emu/g Fe,比饱和磁化强度68.413 4 emu/g,比剩余磁化强度为30.354 1 emu/g,剩磁为19.734 5 Gs.为了检测探针的有效性,进一步使用该探针转染高表达c-erbB2癌基因的SK-Br-3肿瘤细胞株,同时使用正义和无义探针转染SK-Br-3肿瘤细胞株、反义探针转染正常小鼠肝细胞作对照,结果发现反义探针能够特异性进入SK-Br-3肿瘤细胞内,且能明显改善该细胞的磁学参数并降低磁共振扫描下的信号强度. 相似文献
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基因重叠是高等真核生物中一种普遍存在的基因组排现象.综述了高等真核生物中的重叠基因的分类、数量、互补或自然反义转录本(NATs)可能的生物特性和功能及重叠基因与人类疾病的关系. 相似文献