α-干扰素对骨巨细胞瘤细胞凋亡和bFGF及其受体表达的影响

Interferon alfa on Cultured Cells of Giant Cell Tumor of Bone In vitro: Apoptosis and Expression of Basic Fibroblast Growth Factor and Its Receptor

目的:探讨α-干扰素对体外培养的骨巨细胞瘤细胞碱性成纤维细胞生长因子(basicfibroblastgrowthfactor,bFGF)及受体Bek表达与生物学行为的影响,为骨巨细胞瘤术后复发转移的防治探索新途径。方法:采用不同浓度的α-干扰素诱导体外培养的第三代骨巨细胞瘤细胞,观察α-干扰素对瘤细胞生物学行为的影响,流式细胞术与细胞-酶联免疫吸附法分析α-干扰素诱导48h后瘤细胞凋亡情况及Bek/bFGF表达的变化。结果:浓度为1000IU/ml以上的α-干扰素作用10～12h左右,瘤细胞即出现退缩现象,生长明显受到抑制;至48h,瘤细胞数目明显减少,且出现大量细胞碎片。α-干扰素可诱导第三代骨巨细胞瘤细胞发生凋亡,其凋亡率随α-干扰素作用浓度增加而递增,呈明显的剂量依赖关系。α-干扰素可抑制第三代骨巨细胞瘤细胞bFGF和Bek的表达,其抑制强度也随α-干扰素作用浓度的增加而增加,呈明显的量效关系。当α-干扰素作用浓度达到1000IU/ml以上时,其诱导凋亡与抑制Bek/bFGF表达的效应均发生显著性变化。结论:α-干扰素对体外培养的第三代骨巨细胞瘤细胞具有明确的抗肿瘤效应,可作为骨巨细胞瘤术后辅助治疗,以防治和减少其复发转移。

Objective: Giant cell tumor of bone(GCT) is an aggressive, potentially malignant tumor which occurs more frequently in Chinese people than in western people. The study attempts to elucidate the effect of Interferon alfa(IFN-α) on biological behavior and expression of basic fibroblast growth factor (bFGF) and its receptor Bek in cultured cells of GCT in vitro, and to explore an adjuvant therapy for preventing recurrence and pulmonary metastases of GCT. Methods: The fresh specimens of GCT in 15 cases were cultured and the biological behavior of cultured GCT cells were observed in vitro. The third generation cells of GCT incubated with different concentration IFN-α in vitro were observed. The DNA content and apoptosis of the cells treated with IFN-α for 48 hours were analyzed by flow cytometry(FCM), and the expression of bFGF and Bek in these cells were tested by FCM and by cell enzyme-linked immunosorbent assay (C-ELISA). Results: The third generation cells of GCT treated for 10 to 12 hours with IFN-αat the concentration of more than 103IU/ml began to shrink and their growth were significantly inhibited and their numbers decreased. The cells treated for 48 hours with IFN-αdeveloped apoptosis in a dose dependent fashion. IFN-αat the concentration of more than 100IU/ml could significantly inhibited the expression of bFGF and Bek in the cells. The inhibitory effect is also a dose of IFN-αdependent model. Conclusion: IFN-αhas a significant antitumor effect on GCT cells in vitro and may be used as an adjuvant therapy for GCT in order to prevent or decrease the recurrence and metastases.