目的 探讨斑蝥酸钠对食管癌EC9706细胞增殖、迁移及侵袭的影响及机制.方法 将食管癌EC9706细胞随机分为空白对照组、低剂量斑蝥酸钠组、中剂量斑蝥酸钠组、高剂量斑蝥酸钠组及顺铂组,低、中、高剂量斑蝥酸钠组EC9706细胞分别给予终质量浓度1.0、2.5、5.0 mg·L-1的斑蝥酸钠干预,顺铂组EC9706细胞给予终质量浓度140 mg·L-1的顺铂干预,空白对照组使用达尔伯克改良伊格尔培养基培养;采用细胞计数试剂-8法检测各组细胞增殖率,划痕实验检测各组EC9706细胞迁移率,Transwell实验检测各组EC9706细胞侵袭率,实时定量聚合酶链式反应法检测各组EC9706细胞中Wnt3a及β-catenin mRNA水平,Western blot法检测各组EC9706细胞中Wnt3a及β-catenin蛋白水平.结果 培养24、48、72 h,低剂量斑蝥酸钠组、中剂量斑蝥酸钠组、高剂量斑蝥酸钠组和顺铂组EC9706细胞增殖率显著低于空白对照组(P<0.05);低剂量斑蝥酸钠组、中剂量斑蝥酸钠组EC9706细胞增殖率显著高于高剂量斑蝥酸钠组和顺铂组(P<0.05);低剂量斑蝥酸钠组EC9706细胞增殖率显著高于中剂量斑蝥酸钠组(P<0.05);高剂量斑蝥酸钠组与顺铂组EC9706细胞增殖率比较差异无统计学意义(P>0.05);低剂量斑蝥酸钠组、中剂量斑蝥酸钠组、高剂量斑蝥酸钠组和顺铂组EC9706细胞增殖率随着培养时间的延长显著降低(P<0.05).低剂量斑蝥酸钠组、中剂量斑蝥酸钠组、高剂量斑蝥酸钠组和顺铂组EC9706细胞迁移率、侵袭率显著低于空白对照组(P<0.05);低剂量斑蝥酸钠组、中剂量斑蝥酸钠组EC9706细胞迁移率、侵袭率显著高于高剂量斑蝥酸钠组和顺铂组(P<0.05);低剂量斑蝥酸钠组EC9706细胞迁移率、侵袭率显著高于中剂量斑蝥酸钠组(P<0.05);高剂量斑蝥酸钠组与顺铂组EC9706细胞迁移率、侵袭率比较差异无统计学意义(P>0.05).低剂量斑蝥酸钠组、中剂量斑蝥酸钠组、高剂量斑蝥酸钠组和顺铂组EC9706细胞中Wnt3a及β-catenin mRNA和蛋白相对表达量显著低于空白对照组(P<0.05);低剂量斑蝥酸钠组、中剂量斑蝥酸钠组EC9706细胞中Wnt3a及β-catenin mRNA和蛋白相对表达量显著高于高剂量斑蝥酸钠组和顺铂组(P<0.05);低剂量斑蝥酸钠组EC9706细胞中Wnt3a及β-catenin mRNA和蛋白相对表达量显著高于中剂量斑蝥酸钠组(P<0.05);高剂量斑蝥酸钠组与顺铂组EC9706细胞中Wnt3a及β-catenin mRNA和蛋白相对表达量比较差异无统计学意义(P>0.05).结论 斑蝥酸钠能够显著抑制食管癌EC9706细胞的增殖、迁移和侵袭能力,其机制可能与抑制Wnt3a/β-catenin通路有关.
Objective To investigate the effect and mechanism of sodium cantharidate on the proliferation,migration and invasion of esophageal carcinoma EC9706 cells.Methods Esophageal cancer EC9706 cells were randomly divided into blank control group,low-dose sodium cantharidate group,medium-dose sodium cantharidate group,high-dose sodium canthari-date group and cisplatin group.The EC9706 cells in the low-,medium-and high-dose sodium cantharidate groups were given fi-nal mass concentration of 1.0,2.5 and 5.0 mg·L-1 sodium cantharidate intervention,respectively.The EC9706 cells in the cisplatin group were treated with the final mass concentration of 140 mg·L-1 cisplatin,and the cells in the control group was cultured in Dulbecco's modified Eagle's medium.The cell proliferation rate in each group was detected by cell counting reagent-8 method,the mobility of EC9706 cells in each group was detected by scratch test,the invasion rate of EC9706 cells in each group was detected by Transwell method,and the levels of Wnt3a and β-catenin mRNA in EC9706 cells in each group were detected by real-time quantitative polymerase chain reaction method,and the levels of Wnt3a and β-catenin protein in EC9706 cells in each group were detected by Western blot.Results At 24,48 and 72 h of cultivation,the proliferation rate of EC9706 cells in the low-dose sodium cantharidate group,medium-dose sodium cantharidate group,high-dose sodium canthari-date group and cisplatin group was significantly lower than that in the blank control group(P<0.05);the proliferation rate of EC9706 cells in the low-dose sodium cantharidate group and medium-dose of sodium cantharidate group was significantly higher than that in the high-dose sodium cantharidate group and cisplatin group(P<0.05);the proliferation rate of EC9706 cells in the low-dose sodium cantharidate group was significantly higher than that in the medium-dose sodium cantharidate group(P<0.05);there was no significant difference in the proliferation rate of EC9706 cells between the high-dose sodium cantharidate group and cisplatin group(P>0.05);the proliferation rate of EC9706 cells in the low-dose sodium cantharidate group,medium-dose sodium cantharidate group,high-dose sodium cantharidate group and cisplatin group was significantly decreased with the extension of culture time(P<0.05).The mobility rate and invasion rate of EC9706 cells in the low-dose sodium cantharidate group,medium-dose sodium cantharidate group,high-dose sodium cantharidate group and cisplatin group were significantly lower than those in the blank control group(P<0.05);the mobility rate and invasion rate of EC9706 cells in the low-dose sodium cantharidate group and medium-dose sodium cantharidate group were significantly higher than those in the high-dose sodium cantharidate group and cisplatin group(P<0.05);the migration rate and invasion rate of EC9706 cells in the low-dose sodium cantharidate group were significantly higher than those in the medium-dose sodium cantharidate group(P<0.05);there was no significant difference in the mobility rate and invasion rate of EC9706 cells between the high-dose sodium cantharidate group and cisplatin group(P>0.05).The relative expression levels of Wnt3a and β-catenin mRNA and protein in EC9706 cells in the low-dose sodium cantharidate group,medium-dose sodium cantharidate group,high-dose sodium cantharidate group and cisplatin group were significantly lower than those in the blank control group(P<0.05);the relative expression levels of Wnt3a and β-catenin mRNA and protein in EC9706 cells in the low-dose sodium cantharidate group and medium-dose sodium cantharidate group were significantly higher than those in the high-dose sodium cantharidate group and cisplatin group(P<0.05);the relative expressions levels of Wnt3a and β-catenin mRNA and protein in EC9706 cells in the low-dose sodium cantharidate group were significantly higher than those in the medium-dose sodium cantharidate group(P<0.05);there was no significant difference in the relative expression levels of Wnt3a and β-catenin mRNA and protein in EC9706 cells between the high-dose sodium cantharidate group and cisplatin group(P>0.05).Conclusion Sodium cantharidate can significantly inhibit the proliferation,migration and invasion of esophageal carcinoma EC9706 cells,and its mechanism may be related to the inhibition of the Wnt3a/β-catenin pathway.