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作 者:何跃平[1] 张艳[2] 郑家法[1] 申海荣[1] 田梦秋[1] 梁贵玲[1]
机构地区:[1]南华大学附属南华医院耳鼻咽喉科,湖南衡阳421002 [2]南华大学医学微生物学与免疫学教研室
出 处:《南华大学学报(医学版)》2007年第3期389-391,400,共4页Journal of Nanhua University(Medical Edition)
摘 要:目的建立转染人白细胞介素-24(hIL-24)基因的喉癌细胞株,并研究hIL-24的表达情况。方法将携带人白细胞介素-24的质粒pcDNA3.1(+)-hIL-24转导入人喉表皮样癌细胞Hep-2中,G418筛选阳性克隆,RT-PCR、Western blot和ELISA法对hIL-24的表达进行检测。结果IL-24基因成功地转导入Hep-2细胞中并能高效表达。RT-PCR电泳结果显示hIL-24基因在mRNA水平有表达;ELISA法测得106个转染阳性细胞24 h内培养上清液中hIL-24的含量为162.8±15.4 pg/mL,空载体转染及未转染的细胞未检测到hIL-24。结论hIL-24基因成功转染人喉癌细胞系Hep-2细胞且能持续大量表达。Objective To establish human laryngeal cancer cell line transfected with human IL- 24 cDNA and demonstrate its expression in laryngeal cancer cell. Methods The recombinant expression plasmid pcDNA3. 1( + ) -hIL- 24 was transfected into human laryngeal cancer cell line Hep- 2 cells and the positive clone was screened by G418.Then the expression of hIL-24 in Hep- 2 cells was tested by RT-PCR and Western blot assay; the secretion of hIL - 24 in medium was detected by ELISA. Results Human IL- 24 cDNA was successfully integrated into Hep - 2 cells and overexpressed. The result of RT - PCR showed that IL - 24 mRNA was detect- ed, The concentration of hIL - 24 in the supematants of the 106 transfected Hep - 2 cell cultures in 24 hours as detected by ELISA was( 162.8 ± 15.4)pg/mL, no hIL- 24 expression was detected in control group, Conclusion Human IL - 24 cDNA was successfully integrated into Hep - 2 cells and could be expressed with high efficiencv.
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