聚甲基丙烯酸甲酯骨水泥对原代培养脊柱转移瘤细胞的影响和意义

Effect and significance of polymethylmethacrylate bone cement in primary spinal metastasis cells

背景:目前经皮椎体成形技术已广泛应用于治疗老年骨质疏松椎体压缩性骨折以及椎体肿瘤并取得良好疗效,但仍有一些问题需关注和探讨。目的:观察聚甲基丙烯酸甲酯骨水泥对脊柱转移瘤细胞周期变化和细胞凋亡的影响,分析其对脊柱转移瘤的体外抗肿瘤效应。方法:原代培养脊柱转移瘤细胞,将其分为对照组、拉丝期聚甲基丙烯酸甲酯骨水泥组、冷却后聚甲基丙烯酸甲酯骨水泥组,药物处理24h后利用流式细胞术观察凋亡率以及细胞周期,通过SYBRGreenⅠ荧光实时定量-聚合酶链反应法检测肿瘤细胞Bax和Caspase-3mRNA表达的变化。结果与结论:聚甲基丙烯酸甲酯骨水泥有促进脊柱转移瘤细胞凋亡的作用,对照组、拉丝期聚甲基丙烯酸甲酯骨水泥组、冷却后聚甲基丙烯酸甲酯骨水泥组的凋亡率分别为(3.80±1.09)%,(43.46±8.55)%,(28.77±9.39)%,两骨水泥组均能明显提高细胞凋亡率,拉丝期聚甲基丙烯酸甲酯骨水泥组作用更明显(P<0.05)。两骨水泥组均能明显减少G0/G1期细胞,增加S期细胞(P<0.05)。拉丝期聚甲基丙烯酸甲酯骨水泥组作用于脊柱转移瘤细胞后Bax、Caspase-3mRNA的表达显著高于对照组(P<0.05)。提示聚甲基丙烯酸甲酯骨水泥可在体外细胞水平诱导肿瘤细胞凋亡,拉丝期聚甲基丙烯酸甲酯骨水泥作用更加显著。

BACKGROUND： Percutaneous vertebroplasty （PVP） has been widely used and achieved good results in the treatment of senile osteoporotic vertebral compression fracture and vertebral tumor. However, some issues still need further study. OBJECTIVE： To evaluate the effects of polymethylmethacrylate bone cement （PMMA） on the apoptosis and cell cycles of spinal metastasis cells, and to investigate the possible mechanisms of PMMA acting on spinal metastases in vitro. METHODS： After primary culture, spinal metastasis cells were divided into three groups： control group, drawing-off stage PMMA group, cool stage PMMA group. After 24-hour treatment, the apoptosis of spinal metastases cells and its relationship with celt cycles were determined by flow cytometry. The expression of Bax and Caspase-3 mRNA were observed by SYBR green 1 fluorescence quantitative PCR method. RESULTS AND CONCLUSION： The cell apoptosis rates of the control, drawing-off stage PMMA and cool stage PMMA groups were （3.80±1.09）%, （43.46±8.55）%, （28.77±9.39）%, respectively. The latter two groups had significant effect （P 〈 0.05）. Both groups of PMMA remarkably reduced the number of cells in the G0/G1 phase, and increased that in the S phase （P 〈 0.05）. Drawing-off stage PMMA group could markedly increase the mRNA expression of Caspase-3 and Bax as compared with the control group after treatment for 24 hours （P 〈 0.05）. PMMA can significantly induce the apoptosis of spinal metastasis cells and arrest their cell cycles in the S phase in vitro, as well as, with even more significant effect in drawing-off stage than in cool stage.