不同迁移潜能食管癌Ec-109细胞株的建立及生物学特性比较

Establishment of esophageal Ec-109 cell lines with different migration abilities and comparison on their biological characteristics

摘要:

目的：建立高、低迁移能力食管癌Ec-109细胞亚型，并研究其生物学特性差异及其可能机制。方法：改良Transwell小室迁移实验筛选出高、低２种迁移潜能食管癌Ec-109细胞亚群。通过生长曲线、倍增时间检测２种细胞的生长差异；采用Transwell小室检测迁移能力和侵袭的差异；RT-PCR及Western blot检测 RhoC及基质金属蛋白酶－９（matrix metal proteinase-9，MMP-9）表达的差异。结果：改良迁移实验成功筛选出高、低２种迁移潜能食管癌细胞亚群（Ec-109H和Ec-109L），Ec-109H和Ec-109L 食管癌细胞体外生长速度、倍增时间差异无统计学意义（P=0.105）；迁移细胞数分别为（124.9±6.8）和（43.9±6.3）（P=0.000）；侵袭细胞数分别为（50.0±9.7）和（15.2±5.1）（P=0.000）；RT-PCR和Western blot均显示Ec-109H食管癌细胞在基因水平和蛋白水平的 RhoC表达均显著高于Ec-109L食管癌细胞（P=0.000），MMP-9表达差异无统计学意义（P=0.523和 P=0.655）。结论：RhoC在食管癌的迁移转移中发挥重要作用，而MMP-9在食管癌的迁移转移中发挥的作用不明显。

Abstract:

Objective：To establish esophageal Ec-109 cell subtypes with high and low migration abilities and to explore the possible mechanisms of their different biological properties. Methods：Ec-109 cell subtypes with high and low migration abilities were selected out by modified Transwell chamber migration experiment. Growth differences of the two cell types were detected by growth curve and doubling time. Migration and invasion abilities of the two cells were observed by Transwell chamber. Expression of RhoC and matrix metal proteinase-9（MMP-9） in the two cells was detected by RT-PCR and Western blot. Results：Ec-109 cell subtypes with high and low migration abilities（Ec-109H and Ec-109L） were successfully selected out by modified Transwell chamber migration experiment. No statistical difference was observed between Ec-109H and Ec-109L in growth rate in vitro or doubling time（P=0.105）. Num－bers of migrated cells were 124.9±6.8 and 43.9±6.3 respectively（P=0.000）. Numbers of invaded cells were 50.0±9.7 and 15.2±5.1 respectively（P=0.000）. Expression of RhoC was higher in Ec-109H than in Ec-109L demonstrated by RT-PCR and Western blot（P=0.000），but there was no statistical difference in expression of MMP-9 between the two cell types（P=0.523 and P=0.655）. Conclusion：RhoC plays an important role in migration and metastasis of esophageal cancer，while MMP-9 does not.