CAJ | 학술논문

目的：探讨重楼活性单体 PP-26对人结肠癌 SW620细胞增殖抑制作用及其机制。方法：采用噻唑蓝（MTT）法和克隆形成抑制实验观察不同浓度的重楼单体 PP-26对人结肠癌 SW620细胞增殖抑制作用；PI 单染及AnnexinV-FITC ／PI 双染流式细胞术检测细胞周期变化及细胞凋亡水平；Western blotting 检测 PP-26对细胞周期、细胞凋亡相关蛋白以及 Akt 和 ERK 蛋白的表达。结果：与正常肝 LO2细胞相比，重楼单体 PP-26能显著抑制 SW620细胞的生长，作用呈剂量－效应关系；随着 PP-26浓度的增加，细胞克隆形成逐渐减少，与细胞对照组相比有显著差异；不同浓度 PP-26作用后，细胞阻滞于 G1期；PP-26作用细胞24 h 后，CDK4、CDK6表达下降，P15、cyclin D1表达增加；不同浓度 PP-26作用后，细胞晚期凋亡率增加，随浓度增加有上升趋势；PP-26作用细胞24 h 后，线粒体相关凋亡信号通路蛋白 Caspase-9、Caspase-3表达下降，PARP 切割条带增加，细胞促凋亡蛋白 Bax 的表达增加，抗凋亡蛋白 Bcl-2和 Bcl-xL 减少，p-Akt 和 p-ERK 蛋白表达均下降。结论：重楼活性单体 PP-26通过上调 p15促进结肠癌SW620细胞阻滞于 G1期，通过抑制 P13K／Akt 信号通路及 ERK 信号通路，活化线粒体凋亡通路，诱导细胞凋亡。
목적：탐토중루활성단체 PP-26대인결장암 SW620세포증식억제작용급기궤제。방법：채용새서람（MTT）법화극륭형성억제실험관찰불동농도적중루단체 PP-26대인결장암 SW620세포증식억제작용；PI 단염급AnnexinV-FITC ／PI 쌍염류식세포술검측세포주기변화급세포조망수평；Western blotting 검측 PP-26대세포주기、세포조망상관단백이급 Akt 화 ERK 단백적표체。결과：여정상간 LO2세포상비，중루단체 PP-26능현저억제 SW620세포적생장，작용정제량－효응관계；수착 PP-26농도적증가，세포극륭형성축점감소，여세포대조조상비유현저차이；불동농도 PP-26작용후，세포조체우 G1기；PP-26작용세포24 h 후，CDK4、CDK6표체하강，P15、cyclin D1표체증가；불동농도 PP-26작용후，세포만기조망솔증가，수농도증가유상승추세；PP-26작용세포24 h 후，선립체상관조망신호통로단백 Caspase-9、Caspase-3표체하강，PARP 절할조대증가，세포촉조망단백 Bax 적표체증가，항조망단백 Bcl-2화 Bcl-xL 감소，p-Akt 화 p-ERK 단백표체균하강。결론：중루활성단체 PP-26통과상조 p15촉진결장암SW620세포조체우 G1기，통과억제 P13K／Akt 신호통로급 ERK 신호통로，활화선립체조망통로，유도세포조망。
Aim:To investigate the inhibitory effect and apoptotic effects of PP-26,a monomer purified from Paris Polyphylla,on human colorectal cancer SW620 cells.Methods:MTT assay and cell colony formation inhibitory assay were used to determine the inhibitory effects of PP-26 on human colorectal cancer SW620 cells.The percentage of apoptotic cells and cell cycle distribution were determined by flow cytometry.Expressions of cell cycle and apoptosis associated proteins were analyzed by Western blotting. Results:MTT assay showed that PP-26 inhibited colorectal cancer SW620 cells in a dose-dependent manner (P <0.05),but did not inhibit the growth of normal liver LO2 cells at the same concentration. Cell colony formation inhibitory assay demonstrated that cell colonies decreased with the increase of the drug concentrations.PI staining analysis showed that cell cycle was arrested at G1 phase.Western blot-ting analysis showed that the expressions of CDK4 and CDK6 were decreased,while the expressions of P15 and cyclin D1 were increased.Annexin V-FITC /PI staining analysis showed that the percentage of apoptotic cells was increased in a dose-dependent manner after the treatment with PP-26 for 24 h.After the treatment of PP-26 for 24h,the expressions of Caspase-9,and Caspase-3 were downregulated,sug-gesting that the full length protein was cleaved,and the cleaved-PARP was upregulated,the Bcl-2 anti-apoptotic members Bcl-2 and Bcl-xL were downregulated,and the pro-apoptotic protein Bax was upregu-lated,while phospho-Akt,and phospho-ERK were downregulated.Conclusion:PP-26 inhibits cell pro-liferation and induces cell cycle arrested at G1 phase via upregulation p15,and inducing apoptosis through down-regulation of Akt and ERK pathway,and activating the mitochondrial pathway in human colorectal cancer SW620 cells.