麦角胺联合奥沙利铂诱导骨肉瘤细胞增殖和凋亡行为变化

Effect of Ergotamine Combined with Oxaliplatin on Proliferation and Apoptosis of Osteosarcoma Cells

目的探讨麦角胺与奥沙利铂联合应用对骨肉瘤细胞的影响。方法通过使用麦角胺和奥沙利铂分别或联合作用于骨肉瘤细胞,分为空白对照组、麦角胺组、奥沙利铂组和联合用药组。采用细胞增殖实验检测不同处理对骨肉瘤细胞增殖情况的影响;通过克隆形成检测不同处理对骨肉瘤细胞增殖能力的影响;通过细胞凋亡实验检测不同处理对骨肉瘤细胞凋亡行为的影响;Transwell细胞侵袭实验检测不同处理对骨肉瘤细胞侵袭行为的影响;裸鼠成瘤实验检测不同处理对骨肉瘤细胞侵袭行为的影响;Western blot检测PCNA蛋白水平。结果与空白对照组相比,麦角胺、奥沙利铂或联合用药均可抑制骨肉瘤细胞的生长[(1.54±0.23)vs(1.28±0.19)vs(0.78±0.12)vs(0.34±0.08),P<0.01];与空白对照组相比,麦角胺和奥沙利铂处理组中的骨肉瘤细胞集落明显减少,麦角胺和奥沙利铂联合应用时,细胞集落数明显少于单用麦角胺和奥沙利铂[(146.7±17.9)vs(41.2±6.6)vs(38.4±7.1)vs(12.6±4.3),P<0.05];与空白对照组相比,麦角胺和奥沙利铂处理组中的骨肉瘤凋亡细胞数目明显减少,麦角胺和奥沙利铂联合应用时,细胞凋亡数目明显多于单用麦角胺和奥沙利铂[(0.80±0.10)%vs(8.31±0.12)%vs(11.62±1.39)%vs(18.32±3.16)%,P<0.05];与空白对照组相比,麦角胺和奥沙利铂处理组中的骨肉瘤侵袭细胞数目明显减少,麦角胺和奥沙利铂联合应用时,细胞侵袭数目明显少于单用麦角胺和奥沙利铂[(121.3±11.3)vs(68.1±7.6)vs(64.2±7.1)vs(24.6±2.6),P<0.05];与空白对照组相比,麦角胺组裸鼠移植瘤的体积和肿瘤均明显缩小[体积(3.16±0.31)cm^3 vs(0.75±0.13)cm^3,P<0.05;重量(3.08±0.24)g vs(0.69±0.13)g,P<0.05],与空白对照组相比,麦角胺组、奥沙利铂组和联合用药组PCNA蛋白的水平均明显降低[(8.14±0.46)vs(6.32±0.27)vs(3.18±0.19)vs(1.99±0.19),P<0.05]。结论麦角胺可显著促进骨肉瘤细胞的凋亡行为,进而抑制其增殖与侵袭,与药物奥沙利铂联合使用效果更佳。

Objective To investigate the effect of ergotamine combined with oxaliplatin on osteosarcoma cells.Methods Osteosarcoma cells were treated with ergotamine and oxaliplatin,and divided into the control group,the ergotamine group,the oxaliplatin group and combination therapy group. Cell proliferation assay was used to detect the effect of different groups on the proliferation of oxaliplatin cells. The effect of different groups on the proliferation of osteosarcoma cells was detected by cloning. The apoptosis of osteosarcoma cells was detected by apoptosis assay. Transwell cell invasion assay was used to examine the effects of different groups on the invasion behavior of osteosarcoma cells. Nude mouse tumorigenesis assay was used to examine the effects of different groups on osteosarcoma cell invasion. Western blot was used to detect PCNA protein levels. Results Compared with the control group,ergotamine,oxaliplatin or combination inhibited the growth of osteosarcoma cells [（ 1.54±0.23） vs（ 1.28±0.19） vs（ 0.78±0.12） vs（ 0.34±0.08）,P〈0.01]. Compared with the control group,the colonies of osteosarcoma cells in the ergotamine-and oxaliplatin-treated groups were significantly reduced. When ergotamine and oxaliplatin were used in combination,the number of colonies was significantly smaller than when ergotamine or oxaliplatin was used alone[（ 146.7±17.9） vs（ 41.2±6.6） vs（ 38.4±7.1） vs（ 12.6±4.3）,P〈0.05].Compared with the blank control group,the number of apoptotic cells in osteosarcoma in the ergotamine-and oxaliplatin-treated groups was significantly reduced. When ergotamine and oxaliplatin were used in combination,the number of apoptotic cells was significantly larger than when ergotamine was used alone[（ 0.80±0.10） % vs（ 8.31±0.12） % vs（ 11.62±1.39） % vs（ 18.32±3.16） %,P〈0.05].Compared with the control group,the number of osteosarcoma invading cells in the ergotamine-and oxaliplatin-treated groups was significantly reduced. When ergotamine and oxaliplatin were used in combination,the number of cells invaded was significantly smaller than when ergotamine or oxaliplatin was used alone[（ 121.3±11.3） vs（ 68.1±7.6） vs（ 64.2±7.1） vs（ 24.6±2.6）,P〈0.05].Compared with the blank control group,the volume and tumor size of the transplanted tumors in nude mice of the ergotamine group were significantly reduced [（ 3.16± 0.31） vs（ 0. 75 ± 0. 13）,P〈0.05;weight（ 3.08±0.24） vs（ 0.69±0.13）,P〈0.05],and the level of PCNA protein was significantly decreased in the control group,ergotamine group,oxaliplatin group and combination therapy group[（ 8.14± 0.46） vs（ 6.32± 0. 27） vs（ 3. 18 ± 0.19） vs（ 1.99 ±0.19）,P〈0.05].Conclusion Ergotamine can significantly promote the apoptosis of osteosarcoma cells,thus inhibiting their proliferation and invasion. It is even more effective when used in combination with oxaliplatin.