骨髓间充质干细胞促进结肠癌细胞HT29增殖和迁移

Bone marrow-derived mesenchymal stem cells promote proliferation and migration in colorectal cancer cell line HT29

目的探讨骨髓来源的间充质干细胞(bone marrowe-derived mesenchymal stem cells,BD-MSCs)对结肠癌细胞HT29的增殖与迁移能力的影响及其可能的机制。方法将BD-MSCs与结肠癌细胞系HT29共培养,并观察共培养后对结肠癌细胞的增殖和迁移能力的影响。通过Transwell实验检测共培养组与对照组细胞在迁移能力方面的差异;CCK-8实验、克隆形成和成球实验检测HT29细胞的体外增殖能力;体内成瘤实验比较共培养组和对照组细胞的成瘤能力差异;机制的阐明是通过荧光定量PCR和Western blot分别在mRNA和蛋白水平检测共培养组与对照组细胞间E-Cadherin、N-Cadherin、Vimentin、Fibronectin、Snail1、Snail2、Twist1和Twist2的表达差异。结果结肠癌细胞系HT29在与BD-MSCs共培养后,增殖能力显著增强(P<0.05)。与对照组相比,与BD-MSCs共培养后的HT29细胞的迁移能力[(259.6±19.23)个细胞/视野vs(81.6±15.74)个细胞/视野]显著增强(P<0.05)。克隆形成率[(37±2.94)%vs(15.33±2.87)%]和成球率[(31±3.27)%vs(10.33±2.62)%]均显著提高(P<0.05)。在体内成瘤实验中,与BD-MSCs共同注射的实验组所成肿瘤的体积是显著大于对照组的,并且所成肿瘤的重量也是具有显著差异的[(1.73±0.62)g vs(0.54±0.81)g,P<0.05]。最终在mRNA和蛋白水平检测到,与对照组细胞相比,与BD-MSCs共培养的HT29细胞中上皮样标记物E-Cadherin出现显著下调表达,而间质样标记物Vimentin和Twist1则显著上调表达。结论骨髓来源的间充质干细胞促进结肠癌细胞HT29的增殖与迁移。

Objective To investigate the effect of bone marrow-derived mesenchymal stem ceils （BD-MSCs） on the proliferation and migration of colorectal cancer cell line HT29 and to study the possible mechanism. Methods HT29 cells co-cultured with BD-MSCs were defined as a co-culture group, and HT29 cells cultured alone were selected as a control group. The cell migration was detected by Transwell assay. CCK-8 assay, clone formation and sphere formation were applied to determine in vitro proliferation capacity of the HT29 ceils. The tumorigenicity of the co-culture group and the control group was compared by in vivo test. Fluorescence quantitative PCR and Western blotting were used for detecting the expressions of E-cadherin, N-cadherin, vimentin, fibronectin, Snaill, Snail2, Twistl and Twist2 in the 2 groups. Results After co-culture with BD-MSCs, the proliferation of the HT29 cells was significantly increased （P 〈 0. 05）. Compared with the control group, the cell migration of the co-culture group （259.6 ± 19.23 vs 81.6 ± 15.74 cells/field） was significantly enhanced （ P 〈 0. 05 ）, and the clone formation percentage [ （ 37 ±2.94 ） % vs （ 15.33 ± 2. 87） % ] and the sphere formation percentage [ （ 31± 3.27 ） % vs （ 10. 33 ± 2. 62 ） % ] were significantly improved （P 〈 0. 05 ）. The tumor formed in the experimental by injection of HT29 ceils and BD-MSCs was significantly larger than that formed in the control group by injection of HT29 ceils only, and the tumor weight was also statistically different （ 1.73 ±0.62 vs 0. 54 ± 0. 81 g, P 〈 0. 05 ）. The co-culture group exhibited significant down-regulation of E-cadherin and up-regulation of vimentin and Twistl compared with the control group. Conclusion BD-MSCs promote the proliferation and migration in colorectal cancer cell line HT29.