转染反义人BMP2逆转录病毒表达载体对骨肉瘤细胞生长的影响

Growth of human osteosarcoma cells affected by transfection of human antisense BMP2 retrovirus cxpression vcctors

目的 :进一步从分子水平上明确 BMP2在骨肉瘤中的作用机制 ,并为骨肉瘤的基因治疗提供理论依据。方法 :用脂质体方法将构建的 BMP2反义逆转录病毒表达载体转入有 BMP表达的人骨肉瘤细胞OS- 990 1,经 G4 18筛选获得阳性克隆。免疫组化方法检测肿瘤细胞中 BMP和 PCNA的表达 ,并进行流式细胞仪和电镜观察。结果 :转染反义表达载体后肿瘤细胞的 BMP蛋白的表达明显下降 ,并且肿瘤细胞的 PCNA表达降低 ,G1 期细胞比例增加 ,溶酶体增加 ,染色体有断裂。结论 :BMP2反义逆转录病毒表达载体降低骨肉瘤细胞中

objective: To study the effects of BMP on the growth of osteosarcoma cells. Methods: Human antisense BMP2 retrovirus expression vector was constructed and transfected into human osteosarcoma OS 9901 cells by Lipofect AMINE. Positive cell clones were selected with G 418. The expression of BMP and PCNA were determined by immunohistochemical methods, image analysis and expressed as grey level. The morphology and cell cycle distribution of the cells were studied by electronmicroscope and flowcytometry respectively. Results:The grey levels of BMP and PCNA in the transfected cells were 198.4±8.51 and 197.3±3.22, those of the control were 135.1±12.32 and 142.9±8.47,respectively. G 1, G 2 and S phase cells were 51.9%,18.2% and 22.7% in the cell cycle of transfected cells, while those of the control were 52.8%,11.1% and 36.1%, respectively.Increased lysosome and small pieces of chromatin were observed in the transfected cells under electronmicroscope. Conclusion:Transfection of antisense BMP2 may inhibite the expression of BMP and proliferating activity of osteosarcoma cells.