【摘要】 目的　观察白细胞介素 (IL) 7基因体外转染卵巢癌细胞后 ,细胞的增殖特性 ,表面抗原表达和细胞因子分泌的变化 ,及对淋巴因子激活的杀伤细胞 (LAK细胞 )杀伤敏感性的影响。方法　对卵巢上皮性癌细胞株SKOV3进行体外培养 ,将重组质粒pBK CMVIL 7导入SKOV3建立SKOV3 IL 7,将双相表达载体pBK CMV导入SKOV3建立SKOV3 Neo ,应用倒置相差显微镜、四甲基偶氮唑蓝比色试验及流式细胞术 ,观察IL 7基因转染前后 ,SKOV3的增殖特性变化 ;间接标记异硫氰酸荧光素 流式细胞术 ,测定细胞表面抗原表达 ;酶联免疫吸附试验测定细胞因子分泌 ;应用乳酸脱氢酶释放法观察细胞对LAK细胞杀伤敏感性的影响。结果　IL 7基因转染前后 ,SKOV3细胞基本形态、增殖特性及细胞周期分布无明显变化 ;细胞表面人类白细胞抗原ABC(HLA ABC)表达阳性率为 91 8%～99 9% ;人类白细胞抗原DR(HLA DR)未能检测到 ;IL 7基因转染后SKOV3细胞间粘附分子I(ICAM I)表达显著提高 ,SKOV3 IL 7为 (4 0 6± 3 7) % ,SKOV3 Neo为 (2 3 2± 2 7) % ,SKOV3为 (18 9± 7 2 ) % ;转化生长因子 β1 (TGF β1 )表达显著降低 ,SKOV3 IL 7为每 2 4h、10 6 细胞中 (下同 ) 15 8ng L ,SKOV3 Neo为 72 6 μg L ,SKOV3为 396ng L ;IL 7基因转染后 ,SKOV3细胞对LAK细胞?更多还原

【Abstract】 Objective To investigate proliferative activity, expression of cell surface antigens and cytokines of ovarian carcinoma cells transferred by interleukin-7 （IL-7）gene, and its cytotoxic sensitivity to lymphokine activated killer （LAK）cells in vitro through gene transfection technique. Methods Ovarian carcinoma cell line SKOV3 were cultured and transferred by IL-7cDNA.The proliferation activity of IL-7 gene transferred and nontransferred SKOV3 was observed with inverse phase-contrast light microscope, thiazoyl blue tetrazolium bromide assay and flow cytometry. The expressions of cell surface antigen human leucocyte antigen-ABC（HLA-ABC）, human leucocyte antigen-DR（ HLA-DR） and intercellular adhesion molecule-I（ICAM-I） were detected by indirect targeted FITC- flow cytometry. The secretions of IL-2,tumor necrosis factor α（TNF-α）and transforming growth factor（TGF-β₁） were assayed by enzyme linked immunosorbent assay. The cytotoxic sensitivity against LAK was detected by LDH release method. Results The morphology, proliferation,cell cycle distributions and HLA-ABC, HLA-DR expression of IL-7 gene transferred SKOV3 didn’t change after IL-7 gene transfer. The expression of ICAM-I in IL-7 gene transferred SKOV3 significantly increased and TGF-β₁ secretion of IL-7 gene transferred SKOV3 significantly decreased. The cytotoxic sensitivity of IL-7 gene transferred SKOV3 to LAK significantly elevated. Conclusion Ovarian carcinoma cell line SKOV3, after transferred by IL-7 gene, remains unchanged proliferation and apoptosis, upregulation of ICAM-I, downregulation of TGF-β₁ and increases the cytotoxic sensitivity to LAK in vitro.更多还原