Study on transfer of interleukin-7 gene into ovarian carcinoma cells in vitro
- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF OBSTETRICS AND GYNECOLOGY
- Issue:
- 4
- DOI:
- 10.3760/j.issn:0529-567X.2003.04.003
- Key Word:
- 白细胞介素7;卵巢肿瘤;杀伤细胞,淋巴因子激活;细胞因子类
Abstract： 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-CMV IL-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(40.6±3.7)%,SKOV3/Neo为(23.2±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24 h、106细胞中(下同)158 ng/L,SKOV3/Neo为726 μg/L,SKOV3为396 ng/L; IL-7基因转染后,SKOV3细胞对LAK细胞的杀伤敏感性在各效靶比均显著升高. 结论 IL-7基因转染后的SKOV3细胞,不改变卵巢癌细胞的增殖和凋亡特性,但上调ICAM-I表达,下调TGF-β1表达,并使LAK细胞的杀伤敏感性增强.
- [1]钟惠珍,谢幸,谢建华,等.卵巢上皮性癌患者白细胞介素-7测定的临床意义[J].2001,(5).
- [2]程蓓,叶大风,谢幸,等.人白细胞介素7基因克隆及其真核表达载体的构建[J].2002,(3).
- [3]D L, Jicha,J J, Mulé,S A, Rosenberg.Interleukin 7 generates antitumor cytotoxic T lymphocytes against murine sarcomas with efficacy in cellular adoptive immunotherapy.[J].1991,174.
- [4]M\u00f6ller P,Sun Y,Dorbic T,etc.Vaccination with IL-7 gene-modified autologous melanoma cells can enhance the anti-melanoma lytic activity in peripheral blood of patients with a good clinical performance status: a clinical phase I study[J].1998.
- [5]D H, Lynch,R E, Miller.Interleukin 7 promotes long-term in vitro growth of antitumor cytotoxic T lymphocytes with immunotherapeutic efficacy in vivo.[J].1994,179.
- [6]B, Wittig,A, M?rten,T, Dorbic,etc.Therapeutic vaccination against metastatic carcinoma by expression-modulated and immunomodified autologous tumor cells: a first clinical phase I/II trial.[J].2001,12.
- [7]Sherry Mou,Courtney A.Humphries,Kristin L.Komschlies,etc.IL-7 Administration Alters the CD4:CD8 Ratio, Increases T Cell Numbers, and Increases T Cell Function in the Absence of Activation[J].2001,166(5).
- [8]Steven M. Dubinett,Min Huang,Shawkat Dhanani,etc.Down-Regulation of Murine Fibrosarcoma Transforming Growth Factor-β1 Expression by Interleukin 7[J].1995,87(8).
- [9]Neubauer A,Trojaneck B,Schadendorf D,etc.Increase of cytotoxic sensitivity of primary human melanoma cells transfected with the interleukin-7 gene to autologous and allogeneic immunologic effector cells.[J].1997,4(4).