全反式维甲酸对食管癌KYSE70细胞增殖、迁移及Notch1、DLL4、VEGF-C表达的影响
Effect of all-trans retinoic acid on proliferation, migration, expressions of Notch1, DLL4 and VEGF-C in esophageal carcinoma KYSE70 cells

目的:探讨全反式维甲酸(ATRA)单独及联合维甲酸受体β(RARβ)激动剂CD2314或抑制剂LE135对食管癌KYSE70细胞增殖、迁移的影响以及可能机制。方法:将KYSE70细胞分为ATRA组、ATRA+CD2314组、ATRA+LE135组和对照组,采用CCK-8法检测各组细胞分别处理24、48、72 h的增殖情况,通过划痕实验测定处理24 h后的迁移能力,采用Western blot方法检测处理48 h后各组细胞Notch1、DLL4和VEGF-C 蛋白的表达情况。结果:处理24、48和72 h后,ATRA+CD2314组细胞增殖抑制率高于ATRA组,ATRA+LE135组低于ATRA组(P<0.05); 处理24 h后, ATRA组细胞的迁移能力低于对照组,ATRA+CD2314组细胞的迁移能力低于ATRA组(P<0.05),而ATRA+LE135组高于ATRA组(P<0.05); 处理48 h后,ATRA组细胞Notch1蛋白表达量高于对照组,DLL4和VEGF-C蛋白表达量低于对照组,ATRA+CD2314组细胞Notch1蛋白表达量高于ATRA组,而DLL4和VEGF-C蛋白的表达量低于ATRA组; ATRA+LE135组细胞Notch1蛋白表达量低于ATRA组,DLL4和VEGF-C蛋白表达量高于ATRA组(P<0.05)。结论:ATRA可能通过上调Notch1蛋白,下调DLL4、VEGF-C蛋白的表达抑制KYSE70细胞增殖和迁移,增强RARβ的表达可进一步增强这一作用。
Aim: To investigate the effect of ATRA alone or combined with retinoic acid receptor β(RARβ)agonist CD2314 or inhibitor LE135 on the proliferation and migration of esophageal carcinoma KYSE70 cell line and the possible mechanism.Methods:KYSE70 cells were divided into 4 groups, ATRA group, ATRA+CD2314 group, ATRA + LE135 group and control group.CCK-8 assay was used to detect the proliferation of cells in each group after 24, 48, and 72 h treatment. The migration ability of cells in each group was measured by Wound healing assay after 24 h treatment. Western blot was used to detect the expressions of Notch1, DLL4 and VEGF-C after 48 h treatment.Results: After 24, 48 and 72 h treatment, the proliferation inhibition rate of cells in ATRA+CD2314 group was higher than that in ATRA group, which was lower in ATRA+LE135 group than that in ATRA group(P<0.05). After 24 h treatment, the migration ability of cells in ATRA group was lower than that in control group, and the migration ability of cells in ATRA+CD2314 group was significantly lower than that in ATRA group,which was significantly higher in ATRA+LE135 group than that in ATRA group(P<0.05). After 48 h treatment, the expression of Notch1 protein in ATRA group was higher than that in control group, while the expressions of DLL4 and VEGF-C protein were lower; the expression of Notch1 protein in ATRA+CD2314 group was significantly higher than that in ATRA group, while the expressions of DLL4 and VEGF-C protein were significantly lower; the expression of Notch1 protein in ATRA+LE135 group was significantly lower than that in ATRA group, while the expressions of DLL4 and VEGF-C protein were significantly higher(P<0.05).Conclusion: ATRA can inhibit the proliferation and migration of KYSE70 cells by up-regulating Notch1 protein expression and down-regulating the expressions of DLL4 and VEGF-C protein. The increased RARβ expression enhances