食管鳞癌细胞对其相关淋巴管内皮细胞增殖及成环能力的影响

The influence of esophageal squamous cell carcinoma on proliferation and ring formation of the tumor- associated lymphatic endothelial cells

目的观察不同分化程度的食管鳞癌细胞对食管癌微淋巴管内皮细胞体外成环及增殖的影响。方法培养高分化食管鳞癌EC9706细胞及低分化食管鳞癌KYSE150细胞，收集其无血清培养基上清，作为食管癌相关淋巴管内皮细胞的条件培养基；分组三维培养食管癌相关微淋巴管内皮细胞：E1组加入KYSE150条件培养基，E2组加入EC9706条件培养基，K组不更换培养基。计数3组细胞形成的管样结构，并采用细胞计数试剂盒（CCK-8）方法检测各组细胞增殖。结果食管癌相关淋巴管内皮细胞E1组、E2组和K组在3d后成环的数目分别为（34．33±2．08）、（23．33±1．53）、0个，E1〉E2〉K，差异有统计学意义（P〈0．05）；随着时间的延长，E1组和E2组的成环数均有不同程度的减少。CCK-8法检测结果：3组细胞的增殖吸光度（A）值分别为2．657±0．058、2．131±0．045和1．567±0．040，E1〉E2〉K，差异有统计学意义（P〈0．05）。结论不同分化程度的食管癌细胞均可促进食管癌微淋巴管内皮细胞的体外成环及增殖能力，低分化食管癌细胞效应更为明显。

Objective To investigate the effect of esophageal squamous cell carcinoma with different differentiation degrees on proliferation and ring formation of tumor - associated lymphatic endothelial cells in vitro. Methods The well - differentiated esophageal squamous carcinoma EC9706 cells and the poorly- differentiated esophageal squamous carcinoma KYSE150 cells were cultured. The supernatant of culture serum - free medium was colleted as the conditioned medium of esophageal cancer - associated lymphatic endothelial cells. The esophageal cancer - associated micro lymphatic endothelial cells were cultured with three - dimensional culture ： KYSE150 conditioned medium was added into group E1, EC9706 conditioned medium was added into group E2, and medium was not replaced in group K. Tube - like structure of three groups was counted, the tube - like structure in each group was digested, and cell proliferation was detected by cell counting kit - 8 （ CCK - 8 ） method. Results The number of tube - like structure of esophageal cancer associated lymphatic endothelial cells in group E1, group E2 and group K were 34. 33 ±2. 08, 23.33 ±1.53 and 0 respectively after culture for 3 days, and there was significant difference among three groups （ P 〈 0. 05 ）. Along with the extension of time, the ring number of group E1 and group E2 was reduced to varying degrees and there was significant difference among three groups （P 〈 0. 05 ）. CCK - 8 method results revealed that the absorbance （A） values in group E1, group E2 and group K were 2. 657 ±0. 058, 2. 131 ±0. 045 and 1. 567 ±0. 040, respectively. The proliferation rate in group E1 was highest, followed by group E2, and group K, and there was significant difference among three groups （P 〈 0.05 ）. Conclusion Esophageal cancer cells of different differentiation degrees can promote ring formation and proliferation ability of esophageal carcinoma lymphatic endothelial cells in vitro, and the efficacy of low dif- ferentiated esophageal cancer cells is more obvious.