脂质体介导的huIFN-β基因转染对胶质瘤细胞侵袭力抑制作用研究

Inhibitory effects of liposome-mediated gene transfer of interferon-β on invasive potential of human glioma cells in vitro

目的探讨人β-干扰素(huIFN-β)基因脂质体pSV2IFN-β对人脑胶质瘤细胞体外侵袭力的抑制作用。方法用含huIFN-β基因的脂质体体外转染人脑胶质瘤细胞系SHG44,用Western blot和细胞免疫荧光染色法检测目的基因的蛋白表达。通过Boyden小室法检测SHG44转染前后细胞体外侵袭力的变化,同时采用明胶酶谱法分析基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)的比活性。结果转染pSV2IFN-β后,SHG44细胞中的目的基因蛋白有显著表达;磷酸盐缓冲液(PBS)对照组、空载体转染组、pSV2IFN-β脂质体转染组的侵袭性细胞数分别为49.26±14.67、54.63±12.27和18.17±5.42,转染靶基因后SHG44细胞的体外侵袭力受到明显抑制,这与MMPs活性下降的改变相一致。结论转染huIFN-β基因脂质体pSV2IFN-β使脑胶质瘤细胞系SHG44的侵袭力受到明显抑制,提示脂质体介导的huIFN-β基因治疗可能是抗神经胶质瘤侵袭的一个有效的方法。

Objective To evaluate the effects of liposorne-mediated gene transfer of human interferon-β （hulFN-β）on invasion of human glioma cell line SHG44 in vitro. Method SHG44 cells were transfected with pSV2IFN-β in vitro. The protein expression of hulFN-β was detected with Western blotting and immunofluorescence staining respectively. The relative activity of matrix rnetalloproteinases-2 （MMP-2） and matrix rnetalloproteinases-9 （MMP-9）was determined by gelatin zymogram, and the invasion of SHG 44 in vitro was observed using Boyden chamber. Results The protein expression of hulFN-β increased significantly when SHG 44 cells were transfected with pSV2IFN-β. The number of invasion cells of PBS control group, SHG 44 infected with empty liposome and pSV2IFN-β was 49.26 ＋_ 14. 67, 54. 63 ＋_ 12. 27 and 18. 17 ＋ 5.42 respectively. In vitro invasion of glioma cells was significantly inhibited after infected with pSV2IFN-β, and it was consistent with the decline change in MMPs activity. Conclusion The invasion of glioma cells is inhibited significantly after transfection of pSV2IFN-β. These results imply that liposome-mediated gene tmnffer of huIFN-β may be useful for anti-invasion therapy of malignant glioma.