白细胞介素-2基因转导对卵巢癌细胞SKOV3形态及增殖的影响

Interleukin 2 Gene Transfer on the Proliferation and Morphology of Ovarian Cancer Cell Line SKOV3

目的观察白细胞介素２（ＩＬ２）基因转导对卵巢癌细胞镜下形态及增殖性的影响。方法构建携带ＩＬ２基因逆转录病毒载体，将ＩＬ２基因导入卵巢癌细胞系ＳＫＯＶ３，观察细胞光镜下的形态和超微结构的变化，测定细胞核分裂相、生长曲线、３ＨＴｄＲ掺入量及细胞周期。结果以质粒ｐＰＳＲＶ／ＩＬ２为模板，聚合酶链反应（ＰＣＲ）扩增出ＩＬ２ｃＤＮＡ功能编码区，长４９０ｂｐ，插入逆转录病毒载体ｐＬＸＳＮ构建携带ＩＬ２基因载体ｐＬＩＬ２ＳＮ，包装后导入卵巢癌细胞系ＳＫＯＶ３。培养上清液ＩＬ２活性２４小时为３０～３１８Ｕ／１０６细胞。与亲本细胞相比较，光镜下ＩＬ２基因修饰细胞体积增大、形态趋于一致、核浆比缩小；电镜下ＩＬ２基因修饰细胞质高度空泡化，染色质集聚。ＳＫＯＶ３／ＩＬ２细胞核分裂相减少、生长缓慢，倍增时间由３８．４小时延长为６５．４小时，３ＨＴｄＲ掺入量下降５２．３％。细胞周期分析显示，细胞阻滞于Ｇ０／Ｇ１期。结论逆转录病毒可有效介导ＩＬ２基因导入卵巢癌细胞，并长期表达；ＩＬ２基因转导可引起肿瘤细胞形态和超微结构的变化，这种改变可能预示肿瘤细胞生物学特性的变化；ＩＬ２基因转导可抑制ＳＫＯＶ３细胞增?

Objective To study the effect of interleukin 2(IL-2) gene transfer on the proliferation and morphology of ovarian cancer cell line SKOV3. Method IL-2 gene was introduced into ovarian cancer cell line SKOV3 The morphologic changes, mitosis, proliferation curve, 3H uptaking and cell cycle were studied. Results IL 2 cDNA, 490bp, was transduced IL 2 gene expression was detected IL 2 activity was 30～318 U/10 6 cells/24hr. IL 2 gene modified cell is squamas like with smaller nucleus/cytoplasm ratio, lager cell area and more homogeneous. The ultrastructural study showed that the cytoplasma of IL 2 gene modified cells is markedly vacuolized and the chromatin is condensed. There was less mitosis found in SKOV3/IL 2 cells. SKOV3/IL 2 cell proliferate more slowly, the doubling time prolonged from 38.40hr to 65.36hr, 3H uptaking decreased 52.3% of the parent cell. The cell cycle study showed that SKOV3/IL 2 cell was blocked to phase G 1/G 0. Conclusions The retrovirous vector can transduce Il 2 gene into ovarian cancer cell line SKOV3 effectly. IL 2 gene transfer can induce morphologic changs of SKOV3 cells, which may indicate the biologic behavior changes of the targets. IL 2 gene transfer can induce the proliferation inhibition of SKOV3 cells.