hsp702基因在鼻咽癌细胞增殖与凋亡中的作用-role of hsp 702 gene in proliferation and apoptosis of nasopharyngeal carcinoma cells.docx

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hsp702基因在鼻咽癌细胞增殖与凋亡中的作用-role of hsp 702 gene in proliferation and apoptosis of nasopharyngeal carcinoma cells.docx

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hsp702基因在鼻咽癌细胞增殖与凋亡中的作用-role of hsp 702 gene in proliferation and apoptosis of nasopharyngeal carcinoma cells

凋亡百分比，各组间相比无明显变化（P0.05）。另一方面，应用不同浓度的异黄酮类药物 ZSGS-4（0μg/mL、10μg/mL、12.5μg/mL、 15μg/mL 和 17.5μg/mL）分别处理 6-10B 和 5-8F 细胞 12h。各实验组与对照组比较，hsp70-2 蛋白的表达水平明显降低（p0.05）；细胞抑制率明显增强（p0.05）；G0/G1 期和 S 期 DNA 含量明显降低（p0.05）；subG0/G1 期和 G2/M 期 DNA 含量明显升高（p0.05）；细胞凋亡率明显升高（p0.05）。结论：本研究通过上调鼻咽癌细胞 6-10B 的 hsp70-2 蛋白表达，促进其生长增殖；而下调鼻咽癌细胞 6-10B 和 5-8F 的 hsp70-2 蛋白表达，抑制其生长增殖和促使其凋亡。关键词：HSP70-2；鼻咽癌；慢病毒；ZSGS-4ABSTRACTObjective:In this study, the HSP70-2 gene expression of nasopharyngeal carcinoma cell 6-10B could be up-regulated by Lentivirus technology. Then the HSP70-2 gene expression of nasopharyngeal carcinoma cells 6-10B or 5-8F could be down-regulated by isoflavones drugs of ZSGS-4. There should be explored the effects of HSP70-2 gene on the proliferation of nasopharyngeal carcinoma cell.Methods:On the one hand, using of lentiviral technology the product was constructed into the target gene lentiviral vector of pLV-HSP70-2-IRES/eGFP-Neo and the empty lentiviral vector of pLV-CMV-eGFP-Neo. It could get the stably 6-10B cell lines by the target gene lentivirus and the empty lentivirus transducted respectively, i.e.6-10-HSP70-2/eGFP cells (the target gene group) and 6-10B-blank/eGFP cells (the blank control group). On the other hand, the HSP70-2 gene expression of 6-10B or 5-8F cells was down-regulated by isoflavones drugs of ZSGS-4. The hsp70-2 protein expression was analyzed by Western blotting, the proliferation ratios were detected by CCK8 and the cell cycle and the cell apoptosis were analyzed by flow cytometry.Results:In the first place, pEntry221-HSP70-2-IRES/eGFP entry vector was constructed. Then pLV- HSP70-2-IRES/eGFP-Neo lentiviral vector and pLV-CMV-eGFP-Neo empty vector were constructed successfully through LR recombination. In the second place, HSP70-2 lentivirus was packed by 4 lentiviral packaging plasmids system and the clonal cell clusters were screened by G418. hsp70-2 protein expression of 6-10B-HSP70-2/eGFP cells compared with that of 6-10B-blank/eGFP cells or 6-10B ce