黄芪-莪术配伍调控EMT对结肠癌HT-29细胞增殖、迁移和侵袭能力的影响  

Astragali Radix-Curcumae Rhizoma combination inhibits proliferation, migration, and invasion of colon cancer HT-29 cells by regulating EMT

作　　者：杨琦 孙正 朱亦邈 向东阳 张群耀 王访 杨刚 杨灏 唐德才[3] 吴晓宇[3] YANG Qi;SUN Zheng;ZHU Yi-miao;XIANG Dong-yang;ZHANG Qun-yao;WANG Fang;YANG Gang;YANG Hao;TANG De-cai;WU Xiao-yu(the First School of Clinical Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210004,China;School of Chinese Medicine·School of Integrated Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学第一临床医学院,江苏南京210023 [2]南京中医药大学附属医院,江苏南京210004 [3]南京中医药大学中医学院·中西医结合学院,江苏南京210023

出　　处：《中国中药杂志》2023年第3期736-743,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(82174466);江苏省自然科学基金面上项目(BK20211390);江苏省社会发展面上项目(BE2019768);江苏省第六期333高层次人才培养工程项目;江苏省研究生培养创新工程研究生科研与实践创新计划项目(SJCX22-0752)。

摘　　要：观察并探讨黄芪-莪术(AC)配伍调控上皮-间质转化(EMT)对结肠癌HT-29细胞增殖、迁移和侵袭能力的影响。分别用0、3、6、12 g·kg^(-1)AC含药血清作用于人结肠癌HT-29细胞株,采用噻唑蓝(MTT)比色法测定细胞存活和生长,5-溴-2-脱氧尿嘧啶(EdU)细胞增殖实验和Transwell实验检测细胞的增殖、迁移和侵袭能力,流式细胞术检测细胞凋亡情况;将BALB/c雄性裸鼠建立结肠癌皮下移植瘤模型,分为空白对照组、6 g·kg^(-1)AC组、12 g·kg^(-1)AC组,造模后观察记录瘤体的体积与质量,对瘤组织进行HE染色,蛋白免疫印迹(Western blot)检测黄芪-莪术作用后HT-29细胞和裸鼠瘤组织中Bax、caspase-3、cleaved caspase-3相关凋亡蛋白和E-cadherin、MMP9、MMP2、vimentin相关EMT蛋白的表达水平。结果显示,与空白对照组相比,AC作用后的细胞存活率显著降低,且处于增殖期细胞数量明显降低;各给药组的迁移和侵袭细胞数明显下降,细胞凋亡数也明显上升;体内实验中,与空白对照组比,各给药组的瘤体积和瘤体质量均降低,瘤组织呈现出细胞体积缩小,细胞核有固缩现象,表明AC配伍可能改善EMT过程;此外,各给药组的HT-29细胞和肿瘤组织中,Bcl2、E-cadherin蛋白的表达均不同程度上升,Bax、caspase-3、cleaved caspase-3、MMP9、MMP2、vimentin蛋白的表达均不同程度下降。综上,AC配伍能够显著抑制体内外结肠癌HT-29细胞的增殖、侵袭迁移和上皮-间质转化,诱导结肠癌细胞凋亡。This study aims to investigate the effect of Astragali Radix-Curcumae Rhizoma(AC) combination on the proliferation, migration, and invasion of colon cancer HT-29 cells based on epithelial-mesenchymal transition(EMT). HT-29 cells were respectively treated with 0, 3, 6 and 12 g·kg^(-1)AC-containing serum for 48 h. The survival and growth of cells were measured by thiazole blue(MTT) colorimetry, and the proliferation, migration, and invasion of cells were detected by 5-ethynyl-2′-deoxyuridine(EdU) test and Transwell assay. Cell apoptosis was examined by flow cytometry. The BALB/c nude mouse model of subcutaneous colon cancer xenograft was established, and then model mice were classified into blank control group, 6 g·kg^(-1)AC group, and 12 g·kg^(-1)AC group. The tumor weight and volume of mice were recorded, and the histopathological morphology of the tumor was observed based on hematoxylin-eosin(HE) staining. The expression of apoptosis-associated proteins B-cell lymphoma-2-associated X protein(Bax), cysteine-aspartic acid protease-3(caspase-3), and cleaved caspase-3, and EMT-associated proteins E-cadherin, MMP9, MMP2 and vimentin in HT-29 cells and mouse tumor tissues after the treatment of AC was determined by Western blot. The results showed that cell survival rate and the number of cells at proliferation stage decreased compared with those in the blank control group. The number of migrating and invading cells reduced and the number of apoptotic cells increased in the administration groups compared with those in the blank control group. As for the in vivo experiment, compared with the blank control group, the administration groups had small tumors with low mass and shrinkage of cells and karyopycnosis in the tumor tissue, indicating that the AC combination may improve EMT. In addition, the expression of Bcl2 and E-cadherin increased and the expression of Bax, caspase-3, cleaved caspase-3, MMP9, MMP2, and vimentin decreased in HT-29 cells and tumor tissues in each administration group. In summary, the AC com

关 键 词：黄芪-莪术 结肠癌 增殖 侵袭迁移 上皮-间质转化(EMT) 

分 类 号：R285[医药卫生—中药学]