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 and the chemoresistance. Furthermore, the regulatory mechanisms may be related to the increase of the expressions of some stem cell markers in gastric cancer cells.
出处
《中华胃肠外科杂志》
CAS
CSCD
北大核心
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）
