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研究下调骨桥蛋白(osteopontin,OPN)对人U251胶质瘤细胞在裸鼠体内生长的影响并探讨其对胶质瘤生长、侵袭的可能机制.应用RNA干扰技术,将OPN基因的慢病毒干扰载体LV-OPNshRNA感染U251细胞.将对照和试验组U251细胞分别接种裸鼠,建立裸鼠荷瘤模型.3周后测量肿瘤的体积、瘤重并做肿瘤组织病理切片分析;利用RT-PCR和免疫印迹法检测OPN、尿激酶型纤维蛋白酶原激活物(uPA)、基质金属蛋白酶(MMP-2、MMP-9)的mRNA和蛋白表达;免疫组化法检测肿瘤组织微血管密度和血管内皮生长因子(VEGF)表达情况.经OPN的RNA干扰后,能显著降低肿瘤组织OPN mRNA水平及蛋白表达,有效抑制肿瘤细胞生长及侵袭能力,肿瘤体积及重量的减小有统计学意义(P<0.05).感染组uPA、MMP-2和MMP-9的mRNA和蛋白表达明显减少,肿瘤组织的MVD值和VEGF的表达均显著降低.上述结果表明,抑制OPN的表达能明显抑制人U251胶质瘤细胞在裸鼠体内的生长和侵袭,OPN可能通过激活uPA、MMP-2和MMP-9等蛋白酶降解细胞外基质和促进肿瘤血管生成,参与胶质瘤的生长.
To investigate the effect of osteopontin (OPN) on the growth of human U251 glioma cells in nude mice and to explore the possible mechanism of osteopontin (OPN) on the growth and invasion of glioma.Using RNA interference technique, the interference of OPN gene lentivirus The vector LV-OPNshRNA was transfected into U251 cells.U251 cells were inoculated nude mice respectively to establish tumor-bearing model in nude mice.After 3 weeks, the tumor volume and tumor weight were measured and analyzed by RT-PCR and Immunohistochemistry was used to detect the mRNA and protein expression of OPN, uPA and MMP-9. The microvessel density and the expression of vascular endothelial growth factor (P <0.05) .Conclusion: OPN RNA interference can significantly reduce the expression of OPN mRNA and protein in tumor tissue, and effectively inhibit the growth and invasion of tumor cells. The decrease of tumor volume and weight has statistical significance (P <0.05) .The mRNA and protein expressions of uPA, MMP-2 and MMP-9 in the infected group were significantly reduced, while the MVD value and VEGF expression in the tumor tissue were significantly decreased.The above results show that inhibiting the expression of OPN can significantly inhibit the expression of U251 glioma cells In nude mice, OPN may participate in the growth of glioma by degrading extracellular matrix and promoting tumor angiogenesis by activating proteases such as uPA, MMP-2 and MMP-9.