骨髓间充质干细胞调控人胃癌细胞生物学特性的实验研究  

Experimental study of human bone marrow mesenchymal stem cells on regulating the biological characteristics of gastric cancer cells

作　　者：王嘉[1] 俞继卫[1] 吴巨钢[1] 王守练[1] 陈德虎[1] 杨帆[1] 花豹 席小龙 宋书铮 郑林海[1] 姜波健[1] 

机构地区：[1]上海交通大学医学院附属第三人民医院普外一科,201900

出　　处：《中华胃肠外科杂志》2015年第2期159-165,共7页Chinese Journal of Gastrointestinal Surgery

基　　金：上海市卫生局科研课题基金（20134393）;上海交通大学医学院科技基金（13XJ10028）

摘　　要：目的：探讨骨髓间充质干细胞（BMSC）对胃癌细胞生物学特性的影响及其初步调控机制。方法利用Transwell小室构建人胃癌KATO-Ⅲ细胞和BMSC的非接触共培养模型。 CCK-8法检测胃癌细胞增殖能力及对氟尿嘧啶（5-FU）和顺铂的敏感性。 Transwell法检测胃癌细胞侵袭能力。反转录聚合酶链反应法鉴定干细胞标记、凋亡相关因子及上皮-间质转化因子的表达。结果KATO-Ⅲ细胞增殖能力在共培养组中显著强于单独培养组。5-FU和顺铂处理胃癌细胞后，共培养组KATO-Ⅲ细胞生长抑制率显著低于单独培养组（均P＜0．05），且共培养组KATO-Ⅲ细胞抗凋亡基因Bcl-2 mRNA表达升高，促凋亡基因Bax mRNA表达降低（均P＜0．05）。共培养组KATO-Ⅲ细胞穿膜细胞数显著高于单独培养组[（37．33±5．22）比（14．56±2．54），P＜0．01]，且共培养组KATO-Ⅲ细胞上皮-间质转化相关基因Snail、N-cadherin mRNA表达升高，E-cadherin mRNA 表达降低（均P＜0．05）。共培养组KATO-Ⅲ细胞干细胞相关基因CD133、Nanog和Sox-2 mRNA表达均显著高于单独培养组（均P＜0．05）。结论在非接触共培养条件下，BMSC或可诱导胃癌细胞上皮-间质转化，促进细胞增殖能力、侵袭能力及对化疗药物抵抗力。这些调节作用可能与胃癌细胞部分干细胞标记物的上调有关。Objective To investigate the regulatory mechanism of bone marrow mesenchymal stem cells（BMSC） on the biological profiles of KATO-Ⅲcell lines of gastric cancer. Methods Transwell cubicle was applied to build the co-cultured model in non-contact style. The differences of cell proliferation and the resistance of anti-tumour drug （5-fluoropyrimidinedione, 5-FU and Cisplatin, CDDP） between co-cultured group and single cultured group were evaluated by Cell Counting Kit 8-assay （CCK-8）. The invasion ability was detected by Transwell assay. The expressions of stem cell makers, apoptosis-related factors and epithelium-mesenchymal transition （EMT）-related factors were detected by RT-PCR. Results The proliferation ability of KATO-Ⅲ cells in co-cultured group was significantly stronger than that in single cultured group. The growth rate of KATO-Ⅲ cells in co-cultured group was significantly higher than that in single cultured group after treatment of 5-FU and CDDP （P〈0.05）. The mRNA expression level of Bcl-2 was significantly higher in co-cultured group KATO-Ⅲ cells （P〈0.05）, while the mRNA expression level of Bax was significantly lower in co-cultured group KATO-Ⅲ cells（P〈0.05） in comparison with those in single cultured group. As compared to KATO-Ⅲ cells in single cultured group, the number of infiltrating-membrane cells was significantly higher （37.33±5.22 vs 14.56±2.54, P〈0.01） in co-cultured group, and the mRNA expression levels of Snail and N-cadherin were significantly higher in co-cultured group KATO-Ⅲ cells （P〈0.05）, while the mRNA expression level of E-cadherin was significantly lower in co-cultured group KATO-Ⅲ cells （P〈0.05）. The expressions of CD133, Nanog and Sox-2 mRNA in co-cultured group KATO-Ⅲ cells were significantly higher than those in single cultured group （P〈0.05）. Conclusions In co-cultured model sharing non-contact style, BMSC can enhance such properties of KATO-Ⅲ gastric cancer cells as the proliferation, the invasion an

关 键 词：胃癌细胞 骨髓间充质干细胞 上皮-间质转化 生物学特性 

分 类 号：R735.2[医药卫生—肿瘤]