人结直肠肿瘤干细胞致瘤能力及Dvl2蛋白表达的研究  

Tumorigenic ability of colorectal tumor stem cells and the expression of Dv12 protein

作　　者：李利发 张井潇 陈小波[1] 周瑜[3] 罗霞[4] 张广军[1] 周何[3] 周彤[1] 

机构地区：[1]川北医学院附属医院胃肠外科,四川省637000 [2]川北医学院附属医院心内科,四川省637000 [3]川北医学院肝胆胰肠研究所 [4]川北医学院临床医学系

出　　处：《中华普通外科杂志》2016年第9期774-776,共3页Chinese Journal of General Surgery

基　　金：四川省教育厅自然科学基金重点资助项目（14ZA0184）

摘　　要：目的研究人结直肠肿瘤干细胞的致瘤能力及Dvl2蛋白的表达情况。方法以无血清培养液培养人结直肠肿瘤HT29球细胞为基础．通过克隆形成实验和裸鼠皮下成瘤实验，在体内和体外观察HT29细胞系及其球细胞的集落形成和肿瘤生长速度．Westernblotting实验检测Dvl2蛋白在两种细胞中的表达情况。结果HT29球细胞组形成的肿瘤细胞集落数量比HT29细胞组多。HT29球细胞组的裸鼠皮下成瘤率为95％（19／20），高于HT29细胞组的60％（12／20），两组相比差异有统计学意义（X^2=5．16，P〈0．05）；皮下成瘤第4周末，HT29球细胞组的肿瘤体积大于HT29细胞组，两组相比差异有统计学意义[（334±71）mm。比（113±26）mm^2，t=12．33，P〈0．05]；皮下成瘤第8周末，HT29球细胞组的肿瘤体积大于HT29细胞组，两组相比差异有统计学意义[（796±181）rnm。比（318±124）mm^3，t=7．68，P〈0．05]。Dvl2蛋白在HT29细胞中的相对表达量高于在HT29球细胞中的相对表达量，两组相比差异有统计学意义[（0．89±0．60）比（0．37±0．09），t=14．85，P〈0．05]。结论与HT29细胞相比，HT29球细胞具有更强的集落形成和促进肿瘤生长的能力，并且这种较强的致瘤能力可能与Dvl2蛋白的低表达有关。Objective To investigate the tumorigenic ability of human colorectal tumor stem cells, as well as the expression of Dv12 protein in spherical ceils of HT29. Methods Based on serum-free culture cultivate the spherical cells of HT29 in human colorectal tumor, via clone formation assay and subcutaneous tumor experiment. We observed the difference of colony forming and tumor growth rate between two cell lines in vivo and vitro; The expression difference of Dvl2 protein was detected by Western blotting assay in these two cell lines. Results The number of tumor cell colony was significantly more in spherical cells of HT29. In subcutaneous tumor aspect, the spherical cells of HT29 were 95 percent （19/20） vs. 60 percent of HT29 cell line （ 12/20）, （X2 = 5.16, P 〈 0. 05 ）. Tumor volume of HT29 spherical cells was larger than HT29 cell line, in 4 weeks [ （334±71 ） mm^3 vs. （ 113± 26） mm^2, t = 12.33, P 〈 0. 05 ], and in 8 weeks [ （796±181）mm^3 vs. （518±124）mm^3 （t =7.68, P 〈 0. 05） ]. Western blotting assay showed that Dvl2 protein expression in HT29 cells （0. 89±0. 60） was significantly higher than the spherical cells of HT29 （0.37±0.09）, （t = 14.85, P 〈0.05）. Conclusions Compared with HT29 cell line, the spherical cells of HT29 is abler in colony forming and promoting tumor growth, which is partly related with low expression of Dvl2 protein.

关 键 词：结直肠肿瘤 肿瘤干细胞 球细胞 

分 类 号：R735.3[医药卫生—肿瘤]