埃罗替尼与培美曲塞对人胰腺癌细胞BXPC-3及PANC-1生长的作用

Effects of erlotinib and pemetrexed on the proliferation of pancreatic cancer BXPC-3 and PANC-1 cells

目的:探讨埃罗替尼(erlotinib)与培美曲塞(pemetrexed)不同用药时序联合应用对人胰腺癌细胞BXPC-3及PANC-1生长的影响及可能的细胞学机制。方法:RT-PCR法和Western印迹法分别检测2株细胞中表皮生长因子受体(epidemal growth factor receptor,EGFR)mRNA及蛋白的表达;MTT法测细胞增殖;FCM法检测细胞周期。结果:BXPC-3及PANC-1细胞均有EGFR mRNA及蛋白的表达;埃罗替尼(1×10-3～1×10-1mmol/L)和培美曲塞(1.7×10-7～17mmol/L)浓度依赖性抑制2株细胞的生长;二者联合对2株细胞的抑制作用具有协同性,且协同作用强度与给药时序有关:先用培美曲塞24h后序贯用埃罗替尼抑制作用明显强于同时给药(P<0.05)和先用埃罗替尼24h后序贯用培美曲塞(P<0.05)。细胞周期检测结果显示:埃罗替尼联合培美曲塞不同用药顺序对BXPC-3细胞周期未见明显变化(P>0.05)。结论:埃罗替尼与培美曲塞均能抑制人胰腺癌细胞BXPC-3及PANC-1生长,2者联合具协同作用,先用培美曲塞后用埃罗替尼对2株细胞的抑制作用最强,其协同作用与它们对细胞周期的影响无关。

Objective：To investigate the sequence-dependent effect of erlotinib in combination with pemetrexed on the proliferation of human pancreatic carcinoma BXPC-3 and PANC-1 cells, and to explore its cellular mechanism. Methods： The expressions of epidemal growth factor receptor （EGFR） mRNA and protein were examined by RT-PCR and Western blotting, respectively; MTT assay was used to measure the cell proliferation; cell cycle was detected by flow cytometry. Results： EGFR mRNA and protein were expressed in both BXPC-3 and PANC 1 cells. Both erlotinib（10-3 to 10-1 mmol/L）and pemetrexed （1.7×10-7 to 17 mmol/L） significantly inhibited the proliferation of BXPC-3 and PANC-1 cells in a concentration dependent manner. Erlotinib in combination with pemetrexed had synergistic effects in inhibiting the proliferation of BXPC-3 and PANC-1 cells and the synergistic potency was related with the administration sequence. The sequential administration of pemetrexed following erlotinib at 24 h interval had more obvious inhibitory effect on the cell proliferation than simultaneous administration of both drugs and sequential administration of erlotinib following pemetrexed at 24 h interval（P〈0.05）. Cell cycle studies showed that no significant difference of cell cycle distribution was found when erlotinib and pemetrexed were administered concurrently or sequentially （P〉0.05）. Conclusion： Both erlotinib and pemetrexed inhibited the proliferation of BXPC-3 cells and PANC-1 cells. Erlotinib and pemetrexed had synergistic effects in inhibiting cell proliferation. The inhibitory effects were strongest after sequential administration of pemetrexed following erlotinib at 24 h interval. The synergistic effects were not related with their influence on the cell cycle.