免疫抑制剂FK506对结直肠癌细胞增殖、侵袭、迁移能力的影响及作用机制研究  

Effect of immunosuppressant FK506 on proliferation,invasion and migration of colorectal cancer cells and its mechanism

作　　者：姜海波 李卫[1] 姜霞[1] 裴永彬[1] JIANG Hai-Bo;LI Wei;JIANG Xia;PEI Yong-Bin(The Third Department of General,the First Hospital of Hebei Medical University,Shijiazhuang 050031,China)

机构地区：[1]河北医科大学第一医院普外三科

出　　处：《中国免疫学杂志》2019年第20期2488-2493,共6页Chinese Journal of Immunology

基　　金：河北省科技厅医学科学项目(172777164)

摘　　要：目的:探讨免疫抑制剂FK506对结直肠癌细胞增殖、侵袭、迁移能力的影响及作用机制。方法:用不同浓度FK506(0、10、50、100、200、400μg/L)处理HCT116和SW480细胞,采用MTT测定其对肿瘤细胞的增殖抑制率。将HCT116和SW480细胞分为对照组(0μg/L FK506处理)、FK506组1(100μg/L FK506处理)和FK506组2(200μg/L FK506处理),采用流式细胞测定细胞周期,采用Transwell小室测定细胞侵袭、迁移能力,采用Western blot测定细胞周期蛋白D1(Cyclin D1)、基质金属蛋白酶2(MMP2)、MMP9蛋白水平。结果:HCT116和SW480细胞经不同浓度FK506处理后24 h,与对照组比较,FK506浓度≤50μg/L时对HCT116和SW480细胞增殖的抑制作用不明显(P>0.05),FK506浓度>50μg/L时对HCT116和SW480细胞增殖的抑制作用明显增加(P<0.05),且呈现剂量依赖性。与对照组比较,FK506组1和FK506组2 HCT116和SW480细胞G0/G1期升高(P<0.05),HCT116和SW480细胞G2/M期降低(P<0.05),侵袭细胞数和迁移细胞数减少(P<0.05),Cyclin D1、MMP2、MMP9蛋白水平降低(P<0.05);与FK506组1比较,FK506组2 HCT116和SW480细胞G0/G1期升高(P<0.05),HCT116和SW480细胞G2/M期降低(P<0.05),侵袭细胞数和迁移细胞数减少(P<0.05),Cyclin D1、MMP2、MMP9蛋白水平降低(P<0.05)。结论:免疫抑制剂FK506对结直肠癌细胞增殖、侵袭、迁移能力有抑制作用,且呈剂量依赖性,其机制可能与抑制Cyclin D1、MMP2、MMP9的表达有关。Objective:To investigate the effect and mechanism of immunosuppressant FK506 on proliferation,invasion and migration of colorectal cancer cells.Methods: HCT116 and SW480 cells were treated with different concentrations of FK506(0,10,50,100,200,400 μg/L).The inhibition rates of tumor cell proliferation were measured by MTT.The HCT116 and SW480 cells were divided into control group(treatment with 0 μg/L FK506),FK506 group 1(treatment with 100 μg/L FK506) and FK506 group 2(treatment with 200 μg/L FK506).The cell cycles were measured by flow cytometry.Transwell chamber was used to measure cell invasion and migration ability.The levels of Cyclin D1,matrix metalloproteinase 2(MMP2) and MMP9 were determined by Western blot.Results: HCT116 and SW480 cells were treated with different concentrations of FK506 for 24 h,compared with the control group,the inhibitory effect of FK506 concentration ≤50 μg/L on HCT116 and SW480 cell proliferation was not significant(P>0.05),the inhibitory effect of FK506 concentration >50 μg/L on HCT116 and SW480 cell proliferation were significantly increased(P>0.05),and it was dose-dependent.Compared with the control group,the HCT116 and SW480 cells G0/G1 phase in FK506 group 1 and FK506 group 2 were increased(P<0.05),HCT116 and SW480 cells G2/M phase were decreased(P<0.05),the number of invasive cells and migrated cells were decreased(P<0.05),the levels of Cyclin D1,MMP2 and MMP9 protein were decreased(P<0.05).Compared with FK506 group 1,the HCT116 and SW480 cells G0/G1 phase in FK506 group 2 were increased(P<0.05),HCT116 and SW480 cells G2/M phase were decreased(P<0.05),the number of invasive cells and migrated cells were decreased(P<0.05),the Cyclin D1,MMP2 and MMP9 protein levels were decreased(P<0.05).Conclusion: The immunosuppressive agent FK506 has an inhibitory effect on the proliferation,invasion and migration of colorectal cancer cells with a dose-dependent manner,and its mechanism may be related to inhibition of the expression of Cyclin D1,MMP2 and MMP9.

关 键 词：免疫抑制剂 FK506 结直肠癌 增殖 侵袭 迁移 细胞周期蛋白D1 基质金属蛋白酶2 基质金属蛋白酶9 

分 类 号：R605[医药卫生—外科学]