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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 investigate the molecular mechanism of oridonin (ORI) on the proliferation and apoptosis-inducing effects of human osteosarcoma cell lines. Methods The inhibitory effect of ORI on the proliferation of 143B cells was detected by crystal violet staining and Western blot. The effect of ORI on the apoptosis of 143B cells was detected by Annexin V-EGFP staining and Western blot. The expression of ORI in 143B cells was detected by RT-PCR and Western blot Wnt / β-catenin signal; After the overexpression of β-catenin was silenced or overexpressed, crystal violet staining was used to analyze the inhibitory effect of ORI on the proliferation of 143B cells. Results ORI could inhibit the proliferation and induce the apoptosis of 143B cell line (P <0.05), and inhibit the expression of PCNA in 143B cells in a concentration - dependent manner, and at the same time promote the expression of Caspase3 and Bad and suppress the expression of Wnt / β-catenin Signal expression, silencing or overexpression of β-catenin could then promote or inhibit 143B cell apoptosis (P <0.05). Conclusion ORI can inhibit the proliferation and induce the apoptosis of 143B osteosarcoma cell line in a concentration-dependent manner, which may be related to the inhibition of Wnt / β-catenin signaling by ORI.