抑制Src酪氨酸激酶对非小细胞肺癌细胞增殖的影响

Effect of inhibition of Src tyrosine kinase on cell proliferation of NSCLC

目的探讨抑制Src酪氨酸激酶对非小细胞肺癌(non-smalllungcancer,NSCLC)体内外增殖的影响。方法Westernblot和免疫沉淀法检测Src蛋白在NSCLC细胞株中的表达和磷酸化情况;MTT法检测抑制Src酪氨酸激酶活化对NSCLC细胞体外增殖的影响;采用雄性严重联合免疫缺陷(severecombinedimmunitydeficiency,SCID)小鼠,建立肺腺癌PC-9和A549细胞诱导的皮下肿瘤动物模型,研究抑制Src酪氨酸激酶活化对皮下肿瘤增生的影响。结果NSCLC细胞株中Src蛋白明显活化而没有过度表达。Src酪氨酸激酶抑制剂对NSCLC细胞株中Src蛋白的自主磷酸化呈现剂量依赖的抑制作用。亚微摩尔Src酪氨酸激酶抑制剂(<3μmol/L)对PC-9和A549细胞株体外增生表现出剂量依赖性抑制作用,P<0·05。3μmol/LSrc酪氨酸激酶抑制剂也能够显著抑制H226细胞增生,P<0·01。在PC-9细胞诱导的皮下肿瘤模型中,口服Src酪氨酸激酶抑制剂2周后,10和50mg/(kg·d)治疗组对皮下肿瘤体积抑制率分别为73·28%和93·4%,P<0·001。在A549细胞诱导的皮下肿瘤模型中,50mg/(kg·d)治疗3周后差异有统计学意义,抑制率维持在39·4%,P<0·05。结论抑制Src酪氨酸激酶活化能够抑制NSCLC细胞体内外增殖。

OBJECTIVE： To study the effect of inhibition of Src tyrosine kinase on cell proliferation of NSCLC in vitro and in vivo. METHODS： Western blot and immunoprecipitation were used to evaluate the expression and phosphorylation of Src in NSCLC cells. MTT assay was used to examine the effect of inhibition of Src phosphorylation on cell proliferation of NSCLC in vitro. Subcutaneous tumor model induced by PC-9 and A549 cells was used to explore the effect of inhibition of Src tyrosine kinase on cell proliferation of NSCLC in vivo. RESULTS： Src tyrosine kinase was obviously activated, but not over-expression in NSCLC cells. Src tyrosine kinase inhibitor reduced the phosphorylation of Src in a dose-dependent manner in NSCLC cell lines. Submicromolar Src tyrosine kinase inhibitor （〈3 μmol/L） suppressed the proliferation of PC-9 and A549 cells in a dose dependent manner, P〈0.05, and 3 μmol/L Src tyrosine kinase inhibitor also suppressed the proliferation of H226 cells significantly, P〈0.001. For the subcutaneous tumors, after oral feeding with 10 and 50 mg/（kg·d） Src tyrosine kinase inhibitor for 2 weeks, the volume of subcutaneous tumors induced by PC-9 cells were inhibited by 73.28% and 93.4%（P〈0.001）, respectively. While the same treatment was less effective to subcutaneous tumors produced by A549 cells, 50 mg/（kg·d） Src tyrosine kinase inhibitor did not cause significant inhibition until treatment 3 weeks later, and the inhibition rate was about 39.4%, P〈0.05. CONCLUSION： Inhibition of Src tyrosine kinase could suppress the proliferation of NSCLC cells in vitro and in vivo.