番茄红素对体外人骨肉瘤细胞增殖及荷骨肉瘤裸小鼠肿瘤生长的影响

Effects of lycopene on human osteosarcoma cells proliferation in vitro and osteosarcoma growth in nude mice

关键词:  番茄红素  骨肉瘤  增殖  凋亡

Key words:  lycopene  osteosarcoma  proliferation  apoptosis

摘要:

[目的] 研究番茄红素（LP）对体外人骨肉瘤MG-63细胞增殖及荷骨肉瘤裸小鼠肿瘤生长的影响，并初探其机制。[方法] 取对数生长期人骨肉瘤MG-63细胞；设空白对照组，LP（5、10、20 μg/mL）组和顺铂（40 μg/mL）组；给药48 h后，观察细胞形态，检测细胞周期、细胞增值抑制率，观察细胞凋亡状况并测定细胞凋亡率。采用背部皮下注射接种人骨肉瘤细胞的方法建立荷骨肉瘤裸小鼠模型，设模型对照组，LP[5、10、20mg/（kg·d）]和顺铂[2 mg/（kg·d）]，各20只；称量瘤体质量并计算抑瘤率，观察肿瘤组织形态结构变化和细胞凋亡状况，检测肿瘤组织凋亡相关蛋白（bcl-2、Bax、caspase-3）表达。[结果] 体外实验：空白对照组人骨肉瘤MG-63细胞呈贴壁生长、增殖迅速，LP干预组和顺铂组细胞呈现变圆、脱壁、细胞间接触松散状态；与空白对照组比较，LP组和顺铂组细胞周期G₀/G₁期显着延长，细胞增殖抑制率显着升高，凋亡细胞数量明显增多、凋亡率显着升高，差异均具有统计学意义（P<0.05，P<0.01）.体内实验：与模型组比较，LP 10、20 mg/（kg·d）组瘤质量显着减轻、抑瘤率显着升高；LP组肿瘤组织呈片状坏死、瘤细胞皱缩等病理性改变，细胞凋亡指数（AI）显着升高，肿瘤组织bcl-2表达下调、Bax上调而Bax/bcl-2比值显着升高，caspase-3表达显着上调，差异均具有统计学意义（P<0.05，P<0.01）.[结论] LP具有抑制体外人骨肉瘤MG-63细胞增殖和荷骨肉瘤裸小鼠肿瘤生长的药理学作用，其机制可能与LP阻滞细胞周期、调节凋亡相关调控蛋白表达有关。

Abstract:

[Objective] To investigate the effects of lycopene (LP) on osteosarcoma cells proliferation in vitro and osteosarcoma growth in nude mice. [Methods] The human osteosarcoma MG-63 cells in logarithmic growth phase were treated with LP (5, 10, 20 μg/mL) and Cisplatin (40 μg/mL), and set blank group, n=10. Forty eight hours later, the cell morphology and apoptosis were observed. The cellular growth inhibition rate, the cell cycle and apoptosis rate were detected. One hundred nude mice models bearing osteosarcoma were made by injecting human osteosarcoma MG-63 cells were divided randomly into five groups: model control group, LP(5, 10, 20 mg/kg) groups and Cisplatin 2mg/kg group. The tumor weight was detected and the inhibition rate was calculated. The histopathological changes and the tumor cells apoptosis were observed, the expression of bcl-2, Bax, caspase-3 were detected. [Results] In vitro experiment: the cells in blank group showed adherent growth and proliferation rapidly, while the cells in LP treated groups showed that the proliferation was inhibited, the cell number was significantly reduced, and showing retraction, off phenomenon. Compared with blank group, the morphological of LP groups was abnormalities, the cellular growth inhibition rate in LP groups and Cisplatin group were significantly increased, the apoptosis rate was significantly increased (P<0.05, P<0.01). In vivo experiment: compared with model group, the tumor weight of LP 10, 20 mg/(kg·d) groups were significantly decreased and the inhibition rate were significantly increased; necrosis presentation, tumor cell shrinkage and other pathological morphological changes appeared in LP 10, 20 mg/(kg·d) groups; the expression of bcl-2 were significantly decreased, the expression of Bax was significantly increasedm, and the ratio of Bax/bcl-2 were significantly increased, the expression of caspase-3 protein were significantly increased. All of the difference above were significant(P<0.05, P<0.01). [Conclusions] LP can effectively inhibit osteosarcoma cells proliferation in vitro and osteosarcoma growth in nude mice, which perhaps related to its effects of altering the expressing of apoptosis-related proteins.