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| 人白细胞介素—15cDNA真核表达载体的构建及其在肺癌细胞？… | |
| 引用本文： | Shen Y,Cui L,He W,Ba D. 人白细胞介素—15cDNA真核表达载体的构建及其在肺癌细胞？…[J]. 中国医学科学院学报, 1999, 21(5): 368-372 |
| 作者姓名： | Shen Y Cui L He W Ba D |
| 摘 要： | 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－ＳＭ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细胞增殖法阳性细胞ＩＬ－１５的表达活性。结果 获得３个ＰＧ |
| 关 键 词： | 白细胞介素15 cDNA 基因转导 肺癌 载体构建 |
Construction of eukaryotic vector for human interleukin-15 cDNA and its expression in lung carcinoma cell lines |
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| Shen Y,Cui L,He W,Ba D. Construction of eukaryotic vector for human interleukin-15 cDNA and its expression in lung carcinoma cell lines[J]. Acta Academiae Medicinae Sinicae, 1999, 21(5): 368-372 | |
| Authors: | Shen Y Cui L He W Ba D |
| Affiliation: | National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005. |
| Abstract: | OBJECTIVE: Human interleukin(hIL)-15 expression of lung cancer cell lines transfected by rhIL-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 pL-IL-15-SN was transfected into human lung squmosuse carcinoma (PG) cell line and murine lung adenocarcinoma (LA795) cell line, respectively. Positive clones were obtained by the selection in G418 conditioned culture. Bioactivity on rIL-15 was detected by dependant-proliferation of CTLL-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 expression levels of rhIL-15 ranged from 142 to 201 or from 138 to 178U/(ml.10(6)) for positive PG cells or LA795 cells, respectively. CONCLUSIONS: Human and murine lung carcinoma cells transfected with IL-15 cDNA can express hIL-15 with bioactivity. |
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