IL-24 cDNA转染人喉癌细胞系的建立及其表达的研究

Establishment of Human Laryngeal Cancer Cell Line Transfected with Human IL-24 cDNA and Its Expression

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.