汉防已甲素抑制人结肠癌细胞增殖与TGF-β1的关联机制研究  

Relationship between anti-proliferation effect of tetrandrine and TGF-β1 in human colon cancer cells

作　　者：任文艳[1,2] 陈前昭 周林云[1,2] 邵英[1,2] 廖云鹏[1,2] 王涵[1,2] 朱茄慧 何百成[1,2] 

机构地区：[1]重庆医科大学药理学教研室 [2]重庆市生物化学与分子药理学重点实验室,重庆400016

出　　处：《中国药理学通报》2017年第9期1227-1234,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81572226)

摘　　要：目的研究汉防已甲素(tetrandrine,Tet)抑制人结肠癌细胞增殖与TGF-β1的关系。方法利用细胞增殖抑制实验、定量PCR、流式细胞术、Annexin V-EGFP染色以及Western blot实验,分析Tet对HCT116细胞增殖和凋亡的影响;利用定量PCR、Western blot、细胞增殖抑制和免疫荧光实验,分析在结肠癌细胞中Tet抗结肠癌作用与TGF-β1的关系,以及TGF-β1介导Tet抗结肠癌作用可能的分子机制。结果与对照组相比,Tet抑制HCT116细胞增殖、诱导G1阻滞和细胞凋亡;Tet增加TGF-β1在HCT116细胞中的m RNA和蛋白表达水平,TGF-β1增强Tet抑制HCT116细胞增殖和促进其凋亡,抑制TGF-β1明显减弱Tet的这种作用;Tet降低Akt1/2/3的磷酸化水平,抑制PI3K增强Tet对结肠癌细胞增殖的抑制作用;TGF-β1增强Tet对Akt1/2/3磷酸化的抑制作用,而Tet降低Akt1/2/3磷酸化水平的作用可被TGF-β1抑制剂部分逆转;沉默PTEN促进Akt1/2/3磷酸化,但TGF-β1能翻转沉默PTEN对Akt1/2/3磷酸化的促进作用。结论 Tet能抑制结肠癌细胞增殖和促进凋亡,机制可能与其通过促进TGF-β1表达,抑制Akt1/2/3磷酸化有关。Aim To study the relationship between the anti-proliferation effect of tetrandrine（ Tet） and TGF-β1 in human colon cancer cells. Methods Cell viability assay,Western blot,flow cytometry and AnnexinⅤ-EGFP staining were introduced to analyze the anticancer effect of Tet on HCT116 cells. Real-time PCR,Western blot,cell viability and immunofluorescent were employed to determine the relationship between the anti-cancer effect of Tet and TGF-β1 in HCT116 cells,the relationship between the anti-cancer effect of Tet and PI3K/Akt on HCT116 cells,and how TGF-β1 mediated the anti-cancer effect of Tet on HCT116 cells.Results Compared with the control groups,Tet apparently inhibited the proliferation,and induced cell cycle arrest at G1 phase and apoptosis in HCT116 cells. Tet greatly up-regulated the expression of TGF-β1 either the m RNA or protein level,and exogenous expression of TGF-β1 potentiated the anti-cancer effect of Tet in HCT116 cells,while TGF-β1 inhibitor attenuated it notably. Tet decreased the phosphorylation of Akt1/2/3,but no apparent effect was observed on total protein level of Akt1/2; PI3 K inhibitor enhanced the anti-cancer effect of Tet in HCT116 substantially. Exogenous expression of TGF-β1 enhanced the Tet-induced decrease phorphorylation of Akt1/2/3,which was partly reversed by TGF-β1 inhibitor in HCT116 cells. Meanwhile,knockdown of PTEN elevated the level of phorphorylated Akt1/2/3,which was abolished by the exogenous expression of TGF-β1 in HCT116 cells. Conclusion Tet may be a potent candidate drug for colon cancer treatment,and the anti-cancer effect of Tet may be partly mediated by up-regulating TGF-β1 to inactivate PI3K/Akt signal.

关 键 词：汉防已甲素 结肠癌 HCT116细胞 TGF-Β1 PI3K/AKT PTEN 

分 类 号：R-332[医药卫生] R284.1