人白细胞介素-15cDNA真核表达载体的构建及其在肺癌细胞中的表达

Construction of Eukaryotic Vector for Human Interleukin-15 cDNA and Its Expression in Lung Carcinoma Cell Lines

目的观察人ＩＬ－１５ｃＤＮＡ在腺癌细胞内的表达。方法将人ＩＬ－１５ｃＤＮＡ于ＥｃｏＲＩ、ＢａｍＨＩ位点正向克隆到逆转录病毒载体ｐＬＸＳＮ，构建ｐＬ－ＩＬ－１５－ＳＮ的重组质粒。以Ｌｉｐｏｆｅｃｔｉｎ介导将该重组质粒转染到人肺鳞癌细胞（ＰＧ细胞系）和小鼠肺腺癌细胞（ＬＡ７９５细胞系）。经Ｇ４１８条件培养筛选，获得阳性细胞克隆。再经ＣＴＬＬ－２细胞增殖法测阳性细胞ＩＬ－１５的表达活性。结果获得３个ＰＧ细胞和４个ＬＡ７９５细胞阳性克隆，ＩＬ－１５活性测定显示其表达水平在２４ｈ内分别在１４２～２０１或１３８～１７８Ｕ／（ｍｌ·１０６细胞）。结论ＩＬ－１５ｃＤＮＡ转导的人和小鼠肺癌细胞可表达有生物学活性的ＩＬ－１５。

Human interleukin (hIL)-15 expression of lung cancer cell lines trans feeted byrhIL-15 cDNA in vitro was observed. Methods The recombinant plasmid PL-IL-15-SN was established by inserting IL-15 cDNA into vector PLXSN at sites of EcoR I and BamH I. The pLIL-15-SN was trans feeted into human lung squmosuse carcinoma (PG) cell line and murine lungadenocarcinoma (LA795) cell line, respectively. Positive clones were obtained by the selection inG418 conditioned culture. Bioactivity on rIL-15 was detected by depend ant - proliferation ofCTLL-2 cells in vitro. Results Three positive PG and four positive LA795 cell clones were obtained respectively. The results of the assays for hIL-15 bioactivity showed that the expressionlevels of rhIL-15 ranged from 142 to 201 or from 138 to 178U/ (ml. 106) for positive PG cells orLA795 cells, respectively. Conclusions Human and murine lung carcinoma cells transfectedwith IL-15 cDNA can express hIL-15 with bioactivity.