基于PI3K/Akt通路探讨健脾清热活血方含药血清对人结肠癌SW480细胞增殖、凋亡、周期及P-β-catenin表达的影响  

Experiment on Jianpi Qingre Huoxue Decoction Drug Serum Regulates SW480 Cell Proliferation,Apoptosis,Cycle and the Expression of β-Catenin by Altering PI3K/Akt Signal Pathway

作　　者：张涛[1] 陈玉[2] 王小平[3] 平倩[2] 林春李 卓少元[2] ZHANG Tao CHEN Yu WANG Xiao-ping PING Qian LIN Chun-li ZHUO Shao-yuan(Ruikang Affiliated Hospital of Guangxi University of TCM, Nanning 530011, China Guangxi University of TCM, Nanning 530000,China Jiangxi Institute of TCM,Fuzhou 344700,China)

机构地区：[1]广西中医药大学附属瑞康医院消化科,广西南宁530011 [2]广西中医药大学,广西南宁530000 [3]江西中医药高等专科学校,江西抚州344700

出　　处：《中药材》2016年第7期1618-1622,共5页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金面上项目(81260536);广西自然科学基金项目(2013GXNSFAA011916);广西卫生厅重点项目(2011041)

摘　　要：目的:观察健脾清热活血方对人结肠癌SW480细胞凋亡、增殖、周期及P-β-catenin蛋白的影响,探讨该方防治结肠癌可能机制。方法:培养人结肠癌SW480细胞,分别予空白胎牛血清、不同浓度健脾清热活血方含药血清及PI3K阻断剂LY294002干预24 h,MTT法及流式细胞术检测细胞增殖、凋亡及细胞周期,免疫荧光染色法检测P-β-catenin核内转位。结果:健脾清热活血方含药血清组细胞增殖抑制率及凋亡率显著高于空白对照组(P<0.05);同时S期细胞比例显著升高、G1期细胞比例显著降低(P<0.05)。健脾清热活血方含药血清组及LY294002组细胞P-β-catenin以膜表达为主,而空白对照组P-β-catenin以膜表达缺失及核内表达增加为主。结论:健脾清热活血方可能通过调控P-β-catenin核内转录,阻滞SW480细胞周期于G1期,诱导细胞凋亡,抑制细胞增殖,从而发挥防治结肠癌的效用。Objective： To study Jianpi Qingre Huoxue decoction（ JPQRHX） in preventing colon cancer by observing SW480 cells proliferation,apoptosis,cycle and the expression of P-β-catenin,and to research its mechanism. Methods： SW480 cells were incubated with serum containing blank serum,different concentrations of JPQRHX decocition and PI3 K blocking agent LY294002 for 24 h,respectively. Cell proliferation was detected by MTT assay,cell cycle and apoptosis were detected by flow cytometry. The protein translocation of P-β-catenin was assayed by immunofluorescent staining. Results： The inhibitory rate and apoptosis rate in JPQRHX decoction group were higher than control group（ P〈 0. 05）,respectively. S phase cells were increased significantly,and G1 phase cells and LY294002 group cells were decreased significantly（ P〈 0. 05）. The P-β-catenin in JPQRHX decoction groups were mainly expressed in membrane,while the P-β-catenin in the control group was characterized by deletion in membrane and increased in nucleus. Conclusion：JPQRHX decoction has the ability in curing colon cancer,and the mechanism is associated with altering the expression of P-β-catenin in the cells nucleus,blocking SW480 cells cycle at G1 phase,inhibiting SW480 cells proliferation,and inducing SW480 cell apoptosis.

关 键 词：健脾清热活血方 PI3K阻断剂 细胞增殖 细胞凋亡 细胞周期 磷酸化β-连环素蛋白 

分 类 号：R285.5[医药卫生—中药学]