摘要
目的 探讨Transwell体外侵袭模型适用于胆管癌侵袭性研究的实验条件和方法.观察M受体兴奋剂及拈抗剂对胆管癌侵袭力的影响.方法 将不同浓度（0.5×10^5/mL、1.0×10^5/mL、1.5×10^5/mL、2.0×10^5/mL）的胆管癌细胞200μL置入Transwell小室侵袭模型,分别培养12 h、24 h、36 h、48 h,观察不同浓度细胞在不同时间穿透基质凝胶多空滤膜的细胞数.选取最适浓度的胆管癌细胞与匹罗卡品共同加入 Transwell 的上室,调整小室内匹罗卡品浓度分别为0 mmol/L、0.1 mmol/L、0．3mmol／L、0．5mmol／L，培养最适时间后染色计数小室膜上的细胞数。调整浓度分别为0．01mmol／L、0．01mmol／L、0．05mmol／L、0．1mmol／L的阿托品对应匹罗卡品浓度分别为0、0．3mmoL／L、0．3rrmaol／L、0．3mmol／L与胆管癌细胞共同加入Transwell小室培养后染色计数。结果随着作用时间及细胞浓度的增加，穿透基质胶的细胞数随之增多，但当时间超过36h，细胞浓度大于1．0×10^5／mL时增长趋于平稳。加入匹罗卡品后，对照组与实验组比较差异有统计学意义（P〈0．05），实验组高中低浓度之间的差异无统计学意义。单阿托品组与空白对照组比较差异无统计学意义（P〉0．05）。加入匹罗卡品及阿托品后，单阿托品组与实验组比较，差异有统计学意义（P〈0．05），低中高浓度组之间差异无统计学意义。结论36h的侵袭时间和1．0×10^5／mL的细胞浓度最能体现不同药物或试剂对胆管癌细胞的侵袭能力的影响，而受细胞浓度和侵袭时间的影响较小。胆管癌细胞可能存在M受体，并且在胆管癌的浸润和转移中具有重要作用。
Objective To explore the experiment condition and method for the application of in vitro in vasive Transwell chamber and to observe muscarinicreceptor stimulant and muscarinicreceptor antagonist＇s influence to cholangiocarcinoma＇s invasiveness.Methods Two hundred microliter cell suspension of various concentrations（0.5×10^5/mL,1.0×10^5/mL,1.5×10^5/mL and 2.0×10^5/mL）was added into the upper chamber of the Transwell chamber,and the cells were allowed to penetrate the matrigel for 12,18,24and 48 hours respectively.The numbers was gotten as the invasive cells on the under surface of the membrane.After optimal cell concentration and time were gotten,pilocarpine of various concentrations（0 mmol/L,0.1 mmol/L,0.3 mmol/L and 0.5 mmoL/L）was added into the upper chamber of the Transwell chamber,then the cells on the matrigel were stained and counted.So did the cells when atropine of various concentrations（0.01 mmol/L,0.01 mmol/L,0.05 mmoVL and 0.1 mmol/L）were added into the upper chamher of the Transwell chamber in according to pilocarpine of various concentrations（0 mmol/L,0.3 mmol/L,0.3 mmol/L and 0.3mmol/L）.Results With the increase of the time and cell concentrations,the cells couts that penetrated the matrigel increased,while the increase tended to he stable when the culture time exceeded 36 hous and the cell concentration Was over 1.0×10^5/mL.By adding pilocarpine,there were significant differences between the control and experimental groups（P〈0.05）,but there were no significant differences in experimental groups with various concentrations.There were no significant differences in blank group and experimental groups with atropine added（P〉0.05）.When added pilocarpine and atropine,there were significant differences between blank and experimental groups（P〈0.05）,but there were no significant differences in experimental groups with various concentrations.Conclusions Thirty-six hours as invasive time,and one cell concentration 1.0 × 10^5/mL were optimal to test invasion abilities
出处
《国际外科学杂志》
2011年第5期298-301,F0003,共5页
International Journal of Surgery
关键词
Transwell小室
胆管癌
M受体
体外侵袭
Transwell chamber
Cholangiocarcinoma
Muscarinicreceptor
In vitro invasiveness
