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目的:探讨维拉帕米和利血平对肺癌细胞株A549和H460侧群细胞(side population,SP)的阻断差异机制及意义,探讨CD133作为肺癌干细胞表面分子标志的可行性。方法:应用荧光激活细胞分选方法检测和分选A549和H460中的SP和非SP细胞。分别应用维拉帕米、利血平作为SP细胞的阻断剂。应用逆转录聚合酶链反应(RT-PCR)方法检测SP和非SP细胞内ABCG2、MDR-1、CD133的表达情况。结果:在阻断A549和H460的SP细胞对Hoechst33342的外排方面,利血平的阻断效果更好。阻断剂在低浓度加入并提前给药效果较好。A549和H460的SP细胞中ABCG2、MDR-1均高表达。CD133在A549和H460肺癌细胞株的SP内过表达。结论:针对ABCG2编码蛋白的阻断剂或许可以更好的克服肺癌因ABC转运蛋白产生的SP细胞耐药。CD133可能被用来作为肺癌干细胞的分子表面标志。
OBJECTIVE: To investigate the mechanism and significance of verapamil and reserpine on the blocking of side population (SP) of lung cancer cell lines A549 and H460, and to explore the feasibility of using CD133 as a molecular marker on the surface of lung cancer stem cells. Methods: The fluorescence activated cell sorting method was used to detect and sort SP and non-SP cells in A549 and H460 cells. Verapamil and reserpine were used as blockers of SP cells respectively. The expression of ABCG2, MDR-1 and CD133 in SP and non-SP cells was detected by reverse transcription-polymerase chain reaction (RT-PCR). Results: Blocking A549 and H460 SP cells in Hoechst33342 efflux, reserpine blocking effect is better. Blockers added at low concentrations and premedicated better. ABCG2 and MDR-1 were highly expressed in the A549 and H460 SP cells. CD133 is overexpressed in the SP of A549 and H460 lung cancer cell lines. CONCLUSIONS: Blockers against ABCG2-encoded proteins may be better able to overcome the resistance of lung cancer cells to ABC cell-mediated SP cell production. CD133 may be used as a molecular surface marker for lung cancer stem cells.