舒尼替尼与吉西他滨联合及序贯应用对K-RAS突变A549细胞的影响

Combinational and sequential effect of sunitinib and gemcitabine on K-RAS mutant A549 cells

目的研究舒尼替尼与吉西他滨单药、联合及序贯应用对人肺腺癌A549细胞增殖及凋亡的作用及其机制。方法实验分为对照组、舒尼替尼组、吉西他滨组、舒尼替尼序贯吉西他滨组、吉西他滨序贯舒尼替尼组和联合组。A549细胞经舒尼替尼与吉西他滨单药、联合及序贯作用后,MTT法检测细胞增殖;Hoechst 33258染色法检测细胞凋亡形态;流式细胞术检测细胞凋亡及周期分布;Western blotting检测磷酸化细胞外调节蛋白激酶(P-ERK1/2)及磷酸化蛋白激酶B(P-AKT)的表达。结果 A549细胞对舒尼替尼耐药,对吉西他滨敏感。与吉西他滨组相比,吉西他滨序贯舒尼替尼组A549细胞增殖抑制率及凋亡率明显增高(P<0.05)。舒尼替尼与吉西他滨分别阻滞A549细胞于G1期及S期。与对照组相比,舒尼替尼序贯吉西他滨组G1期细胞增多(P<0.05),S期细胞减少(P<0.05)。与舒尼替尼组相比,吉西他滨序贯舒尼替尼组A549细胞P-ERK1/2与P-AKT的表达降低(P<0.05)。结论吉西他滨序贯舒尼替尼作用于A549细胞具有协同作用,其机制与酪氨酸激酶受体信号通路的表达有关。

Objective To study the effect of monotherapeutic, combinational and sequential applications of sunitinib and gemcitabine on the proliferation and apoptusis of human lung adenocarcinoma A549 cells and explore its mechanism. Methods The cells were divided into the control group, sunitinib group, gemcitabine group, gemcitabine following sunitinib group, sunitinib following gemcitabine group, and combination group. After A549 cells were treated with monotherapeutic, combinational and sequential applications of sunitinib and gemcitabine, the cell growth inhibitory rate was measured by MTT assay; cellular apoptotic morphology changes were detected by Hoechst 33258 staining; cell cycle and apoptosis rate were evaluated by flow cytometry; the expressions of phosphorylation extracellular regulated protein kinase （P-ERK1/2） and phosphorylation protein kinase B （P-AKT） were detected by Western blotting. Results A549 cells were resistant to sunitinib while sensitive to gemcitabine. The growth inhibitory and apoptosis rate in sunitinib following gemcitabine group were higher than those in gemcitabine group （ P 〈 0.05 ）. Gemcitabine and sunitinib mainly blocked A549 cells on the S phase and Gj phase respectively. The cells in G1 phase of sunitinib following gemcitabine group increased while the cells in S phase decreased than those in the control group （ P 〈 0.05 ）. Expressions of P-ERK1/2 and P-AKT were further inhibited in sunitinib following gemcitabine group than sunitinib group （P 〈 0.05 ）. Conclusion The application of sunitinib following gemcitabine on A549 cells has synergistic effect. The mechanism is related to the expression of tyrosine kinase receptor signal pathway.