Objective Nucleostemin （NS） is a GTP-conjugated protein located in the nucleoli of stem cells and some cancer cells, and maintains cell self-renewal. We aimed to evaluate NS as a potential target for lung carcinoma ...Objective Nucleostemin （NS） is a GTP-conjugated protein located in the nucleoli of stem cells and some cancer cells, and maintains cell self-renewal. We aimed to evaluate NS as a potential target for lung carcinoma gene therapy by investigating NS gene expression and its effect on A549 cell proliferation. Methods NS mRNA and protein expression in A549, HepG2, SMMC-7721, HeLa, and U251 cells was analyzed by RT-PCR and western blotting following transfection of NS siRNAs and negative control siRNA （NC）. The effect on cell proliferation was also analyzed by MTF assays. Results NS mRNA and protein were both expressed in A549 cells and four other tumor cell lines; the relative expression levels were similar in all five cell lines. The three pairs of NS siRNA, either transfected alone or cotransfected into A549 cells, could effectively inhibit the expression of NS mRNA and protein. Moreover, the interference ratio showed an obvious concentration-dependent relationship. NS siRNA treatment resulted in significant inhibition of A549 cell proliferation by 35.7%. Conclusion NS gene was not only highly expressed but also played an important role in A549 cell proliferation. Thus, targeting of NS may be a promising novel strategy for the treatment of lung carcinoma.展开更多
目的探讨核干因子(nucleostemin gene,NS)基因在前列腺癌(prostate cancer,PCa)组织中的表达及其与血清前列腺特异性抗原(prostate specific antigen,PSA)的关系。方法采用逆转录-聚合酶链反应(RT-PCR)及免疫组织化学法检测NS在PCa及良...目的探讨核干因子(nucleostemin gene,NS)基因在前列腺癌(prostate cancer,PCa)组织中的表达及其与血清前列腺特异性抗原(prostate specific antigen,PSA)的关系。方法采用逆转录-聚合酶链反应(RT-PCR)及免疫组织化学法检测NS在PCa及良性前列腺增生(BPH)组织中的表达,分析其在PCa中的表达水平与血清PSA的关系。结果45例PCa组织标本中,NS mRNA的阳性表达率为80.0%(36/45)。20例BPH组织中均未见阳性表达。在高分化、中分化、低分化PCa组织中NS蛋白阳性表达例数分别为4、19、13例。NS蛋白表达与PCa恶性程度呈正相关(r=0.954,P<0.05),与患者血清PSA水平无明显相关性(r=0.237,P>0.05)。结论NS蛋白表达与PCa的分级密切相关,可作为一种诊断PCa的新肿瘤标志物。展开更多
A tumor-targeting gene vector G250mAb-PEI-PEG has been prepared by modification of polyethylenimine (PEI) with polyethyleneglycol (PEG) and G250, a monoclonal antibody against the G250 antigen on tumor cell surface. T...A tumor-targeting gene vector G250mAb-PEI-PEG has been prepared by modification of polyethylenimine (PEI) with polyethyleneglycol (PEG) and G250, a monoclonal antibody against the G250 antigen on tumor cell surface. The transfection efficiency was as high as 70% in G250 positive HeLa cells, whereas the transfection efficiency was relatively low (30%) in normal NIH3T3 cells. A plasmid encoding the short hairpin RNA (shRNA) specific for nucleostemin gene (NS) was efficiently transfected into the HeLa cells with this nonviral gene vector. RNA interference down-regulated the expression of NS gene in HeLa cells, inhibited cells proliferation and induced apoptosis. However, the growth and activity of the NIH3T3 cells were not affected under the same treatment. These results indicate that the reported nonviral gene vector, G250mAb-PEI-PEG, can target and efficiently deliver genes into HeLa cells, and has the potential for the cervical cancer treatment.展开更多
基金supported by the Plan of Promoting Sichuan University science research start up fund (NO.0082204127092)
文摘Objective Nucleostemin （NS） is a GTP-conjugated protein located in the nucleoli of stem cells and some cancer cells, and maintains cell self-renewal. We aimed to evaluate NS as a potential target for lung carcinoma gene therapy by investigating NS gene expression and its effect on A549 cell proliferation. Methods NS mRNA and protein expression in A549, HepG2, SMMC-7721, HeLa, and U251 cells was analyzed by RT-PCR and western blotting following transfection of NS siRNAs and negative control siRNA （NC）. The effect on cell proliferation was also analyzed by MTF assays. Results NS mRNA and protein were both expressed in A549 cells and four other tumor cell lines; the relative expression levels were similar in all five cell lines. The three pairs of NS siRNA, either transfected alone or cotransfected into A549 cells, could effectively inhibit the expression of NS mRNA and protein. Moreover, the interference ratio showed an obvious concentration-dependent relationship. NS siRNA treatment resulted in significant inhibition of A549 cell proliferation by 35.7%. Conclusion NS gene was not only highly expressed but also played an important role in A549 cell proliferation. Thus, targeting of NS may be a promising novel strategy for the treatment of lung carcinoma.
基金Supported by the National Natural Science Foundation of China (Grant No. 20774050)National Natural Science Funds for Distinguished Young Scholar (Grant No. 30725030)National Basic Research Program of China (Grant No. 2009CB918900-G)
文摘A tumor-targeting gene vector G250mAb-PEI-PEG has been prepared by modification of polyethylenimine (PEI) with polyethyleneglycol (PEG) and G250, a monoclonal antibody against the G250 antigen on tumor cell surface. The transfection efficiency was as high as 70% in G250 positive HeLa cells, whereas the transfection efficiency was relatively low (30%) in normal NIH3T3 cells. A plasmid encoding the short hairpin RNA (shRNA) specific for nucleostemin gene (NS) was efficiently transfected into the HeLa cells with this nonviral gene vector. RNA interference down-regulated the expression of NS gene in HeLa cells, inhibited cells proliferation and induced apoptosis. However, the growth and activity of the NIH3T3 cells were not affected under the same treatment. These results indicate that the reported nonviral gene vector, G250mAb-PEI-PEG, can target and efficiently deliver genes into HeLa cells, and has the potential for the cervical cancer treatment.
