阿仑膦酸钠抑制骨巨细胞瘤细胞生长和诱导其凋亡的研究

目的]骨巨细胞瘤是一种潜在的恶性病变，具有手术后易复发的特点。二膦酸盐是抗骨质疏松药，可以抑制破骨细胞性骨吸收，近来发现其还有抗肿瘤作用。本研究是探讨第3代二膦酸盐——阿仑膦酸钠是否能够抑制骨巨细胞瘤细胞生长，诱导骨巨细胞瘤细胞凋亡，探讨应用二膦酸盐能否成为一个防止骨巨细胞瘤复发的方法。方法]在体外培养骨巨细胞瘤细胞，给予不同浓度的阿仑膦酸钠，作用不同时间后，应用M1Tr法检测骨巨细胞瘤细胞的活性是否受到抑制，TUNEL染色法观察骨巨细胞瘤细胞经药物作用后是否发生凋亡，流式细胞术检测凋亡率，观察药物作用后凋亡蛋白Caspase-3活性的表达是否增加。结果]经阿仑膦酸钠作用后瘤细胞活性减低，可以发现阿仑膦酸钠抑制骨巨细胞瘤细胞生长的作用可以随时间和浓度的增加而增高。TUNEL法观察到瘤细胞凋亡染色阳性，流式细胞仪检测阿仑膦酸钠作用后骨巨细胞瘤细胞的凋亡率也随着时间和浓度的增加而增高。进一步检测随着阿仑膦酸钠浓度的提高，骨巨细胞瘤细胞的Caspase-3活性表达也增加。结论]阿仑膦酸钠对于体外培养的骨巨细胞瘤细胞的活性有抑制作用，可以抑制其生长并诱导肿瘤细胞内的Caspase-3活性表达，促其凋亡，阿仑膦酸钠可能成为治疗和预防骨巨细胞瘤复发的一个治疗方法，但还需要进一步的体内实验研究。

Study of the Alendronate inhibiting growth and inducing apoptosis of giant cell tumor of bone in vitro

Objective]Giant cell tumor of bone is notorious for its local aggressive behavior and its tendency to recur after operative treatment.Bisphosphonates is the drug of anti-osteoporosis.It is found also have anti-cancer effect recently.We conducted experiment examing the effect of bisphosphonates alendronate on the growth and survival of the cells.To study if bisphosphonates are capable of inducing cells death and significantly inhibiting their growth in vitro.Method]Cells viability was detected by MTT Assay after the tumor cells were cured with different concentration and different time.Tumor cells apoptosis with in situ TUNEL assay and flow cytometry was detected.The active Caspase-3 was also detected.Result]After exposure to alendronate,the cells exhibited the characteristic features of cell shrinkage,rounding and partial detachment,and demonstrated the lobulated appearance of apoptotic cells.It was much more prominent while the treating time prolonged or the concentration increased.Alendronate((5 200) M) treatment for 24 h,resulted in 2.79%~31.17% decrease in cell viability,and 11.13%~49.94% for 72 h,respectively.A significant dose-dependent and time-dependent decrease in the number of viable cells was observed in the GCT cells.After Alendronate treat for 24 h,the mean cell population in apoptosis was 14.32% at concentration 5 mmol/l,and 40.24% at 200 mmol/l.It was up to 18.41% and 42.22% respectively after 48 h.In Alendronate-treated GCT cells,Caspase-3 activation was observed.The cell response varied with doses of Alendronate showing the levels of Caspase-3 expression with a dose dependent response.Conclusion]In conclusion,we demonstrated that bisphosphonate alendronate could inhibit GCT cells in the present study.This response was time-dependent and dose-dependent.Alendronate inducing apoptosis in GCT cells is mediated by the activation of Caspase-3.