启膈散对食管癌细胞间同质黏附力的影响

摘要

Background The incidence of esophageal cancer has distinct geographical characteristics, with approximately 50% of all newly diagnosed cases occurring in China annually. Objective To investigate the effects and mechanism of Qigesan on the cell - cell adhesion of esophageal carcinoma. Methods From March to June in 2015,the human esophageal carcinoma cell lines Eca109,TE13 and TE1 were cultured in vitro. Eca109,TE13 and TE1 were treated with the different dose of Qigesan(0 μg/ ml in control group,50 μg/ ml in low dose group,100 μg/ ml in high dose group),the cell - cell adhesion was tested by mechanical method,the expression level of Cx32 in Eca109,TE13 and TE1 was detected by Western blotting method. The expression of E - cadherin in Eca109,TE13 and TE1 with different dose of Qigesan(0 μg/ ml in control group,100μg/ ml in Qigesan group)was detected by SP immunohistochemistry method. The metastatic abilities of Eca109,TE13 and TE1 with different dose of Qigesan(0 μg/ ml in control group,50 μg/ ml in low dose group,100 μg/ ml in high dose group)were detected by scratch - wound assays. Results The cell - cell adhesion of Eca109,TE13 and TE1 in low dose group and high dose group was higher than that in control group respectively(P < 0. 05);the cell - cell adhesion of Eca109,TE13 and TE1 in high dose group was higher than that in low dose group(P < 0. 05). The expression level of Cx32 of TE1 in low dose group was higher than that in control group(P < 0. 05);the expression level of Cx32 of Eca109,TE13 and TE1 in high dose group was higher than that in control group and low dose group respectively(P < 0. 05). The expression of the E - cadherin was negative in control group and Qigesan group. At 12 h and 24 h,the scratch of Eca109,TE13 and TE1 in low dose group and high dose group was wider than that in control group respectively(P < 0. 05). At 12 h,the scratch of Eca109 in high dose group was wider than that in low dose group(P < 0. 05). At 12 h and 24 h,the scratch of TE13 and TE1 in high dose group was wider than that in low dose group(P < 0. 05). Conclusion Qigesan can improve the expression level of Cx32,then improve the cell - cell adhesive ability and reduce the migratory ability of esophageal carcinoma cells.%背景食管癌的发病率有明显的地理特征，每年约有50%的新诊断病例发生在中国。目的研究启膈散对食管癌细胞间同质黏附力的影响，并探讨其作用机制。方法2015年3—6月，体外培养 Eca109、TE13、TE1，采用机械法检测不同剂量启膈散（0、50、100μg/ ml）（分别为对照组、低剂量组、高剂量组）干预后 Eca109、TE13、TE1同质黏附值，Western blotting 法检测不同剂量启膈散（0、50、100μg/ ml）（分别为对照组、低剂量组、高剂量组）干预后 Eca109、TE13、TE1中 Cx32表达水平，免疫细胞化学法检测不同剂量启膈散（0μg/ ml 和100μg/ ml）（分别为对照组、启膈散组）干预后 Eca109、TE13、TE1中 E - cadherin 表达情况，划痕实验检测不同剂量启膈散（0、50、100μg/ ml）（分别为对照组、低剂量组、高剂量组）干预后 Eca109、TE13、TE1迁移能力。结果低剂量组、高剂量组 Eca109、TE13、TE1同质黏附值大于对照组（P ＜0.05）；高剂量组 Eca109、TE13、TE1同质黏附值大于低剂量组（P ＜0.05）。低剂量组 TE1中 Cx32表达水平高于对照组（P ＜0.05）；高剂量组 Eca109、TE13、TE1中 Cx32表达水平高于对照组、低剂量组（P ＜0.05）。对照组、启膈散组 Eca109、TE1、TE13中 E - cadherin 表达均为阴性，未见表达增强。低剂量组、高剂量组12 h、24 h时 Eca109、TE13、TE1划痕宽度大于对照组（ P ＜0.05）；高剂量组12 h时Eca109划痕宽度大于低剂量组，12 h、24 h时 TE13、TE1划痕宽度大于低剂量组（P ＜0.05）。结论启膈散能够增加Cx32表达水平，从而提高食管癌细胞间同质黏附力，进而抑制食管癌细胞的迁移。