冬凌草甲素促进骨肉瘤细胞143B凋亡及其可能机制

Oridonin induces apoptosis of human osteosarcoma cell line 143B and related mechanism

目的研究冬凌草甲素(oridonin,ORI)对人骨肉瘤细胞株增殖抑制和凋亡诱导作用的分子机制。方法以结晶紫染色和Western blot检测ORI对143B细胞增殖的抑制作用,Annexin V-EGFP染色法和Western blot检测ORI对143B细胞凋亡的促进作用;RT-PCR和Western blot检测ORI对143B细胞内Wnt/β-catenin信号的影响;沉默或过表达β-catenin后,结晶紫染色分析ORI对143B细胞增殖的抑制作用的变化。结果 ORI能够抑制143B细胞株增殖,并诱导其凋亡(P<0.05);呈浓度依赖性抑制143B细胞PCNA的表达,同时促进其Caspase3、Bad的表达;能够抑制143B细胞内Wnt/β-catenin信号的表达,沉默或过表达β-catenin后则可以相应地促进或抑制143B细胞的凋亡(P<0.05)。结论ORI能够浓度依赖性抑制143B骨肉瘤细胞株增殖并诱导其凋亡,可能与ORI能够抑制其Wnt/β-catenin信号转导有关。

Objective To study the molecular mechanism of oridonin （ORI） in inhibiting proliferation and inducing apoptosis of human osteosarcoma cell line 143B. Methods 143B cells were treated with different doses of ORI in a certain time before being used in the experiment. The proliferation inhibitory effect of ORI was tested by crystal violet staining and Western blotting. The effect of inducing apoptosis was detected by Annexin V-EGFP staining and Western blotting. The influence on WNT/β-catenin signal pathway was investigated by RT-PCR and Western blotting. After Si-β-catenin or Ad-β-catenin was used to silence or over-express β-catenin in 143B cells, the proliferation inhibitory effect of ORI was tested by crystal violet staining again. Results ORI could inhibit the proliferation and induce apoptosis of 143B cells in a concentration-dependent manner. It also suppressed the expression of proliferating cell nuclear antigen （PCNA） , promoted the expression of Caspase 3 and Bad, and decreased the level of β-eatenin. The apoptosis of 143B cells was promoted or inhibited by silencing or over-expressing β-eatenin. Conclusion ORI can inhibit the proliferation and induce the apoptosis of 143B ceils in a concentration-dependent manner, which may be associated with the down-regulation of 13-catenin.