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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 investigate the effect of inhibition of Src tyrosine kinase on the proliferation of non-small-lung cancer (NSCLC) in vitro and in vivo. Methods Western blot and immunoprecipitation were used to detect the expression and phosphorylation of Src protein in NSCLC cell line. MTT assay was used to detect the effect of Src tyrosine kinase activation on proliferation of NSCLC cells in vitro. Male severe combined immunodeficiency (SCID) was used. In mice, an animal model of subcutaneous tumor induced by lung adenocarcinoma PC-9 and A549 cells was established to investigate the effect of inhibition of Src tyrosine kinase activation on subcutaneous tumor proliferation. Results The Src protein was significantly activated but not overexpressed in the NSCLC cell line. Src tyrosine kinase inhibitors exhibit dose-dependent inhibition of autonomous phosphorylation of Src protein in NSCLC cell lines. Submicromolar Src tyrosine kinase inhibitors (< 3 μmol/L) showed a dose-dependent inhibition of the proliferation of PC-9 and A549 cells in vitro, P<0.05. 3 μmol/LSrc tyrosine kinase inhibitors also Can significantly inhibit the proliferation of H226 cells, P <0.01. In the subcutaneous tumor model induced by PC-9 cells, after oral administration of Src tyrosine kinase inhibitor for 2 weeks, the inhibition rates of subcutaneous tumor volume in the 10 and 50 mg/(kg·d) treatment groups were 73·28% and 93·2·, respectively. 4%, P<0.001. In the subcutaneous tumor model induced by A549 cells, the difference was statistically significant after 3 weeks of treatment at 50 mg/(kg·d), and the inhibition rate was maintained at 39·4%, P<0.05. Conclusion The inhibition of Src tyrosine kinase activation can inhibit the proliferation of NSCLC cells in vitro and in vivo.