敲低乙酰辅酶A合成酶短链家族成员2(ACSS2)抑制营养缺乏诱导的宫颈癌细胞迁移和侵袭及机制

Knockdown of ACSS2 inhibits invasion and migration of cervical cancer cells induced by nutrient stress and its mechanism

目的探讨营养缺乏诱导的乙酰辅酶A合成酶短链家族成员2(ACSS2)表达上调对宫颈癌细胞侵袭和迁移能力的作用机制.方法免疫组织化学染色法检测井比较20对宫颈癌组织及癌旁组织中ACSS2的表达差异.培养宫颈癌HeLa细胞和HCvEpC宫颈正常上皮细胞,将培养血清由100 ml/L降至10 mL/L进行营养缺乏处理.Western blot法检测细胞ACSS2及GSK-3β、p-GSK-3β、β-catenin、E-actin、E-cadherin、vimentin的蛋白水平.小干扰RNA(siRNA)靶向下调ACSS2表达,划痕实验检测细胞迁移能力的改变,Transwell^TM实验观察细胞侵袭能力.结果与癌旁正常组织相比,宫颈癌瘤体组织中ACSS2表达较高;营养缺乏处理可上调宫颈癌HeLa细胞ACSS2蛋白水平,HCvEpC宫颈正常上皮细胞未见明显变化;营养缺乏处理可以促进HeLa细胞上皮间质转化(EMT)发生,靶向抑制ACSS2可有效逆转这一进程;营养缺乏处理促使HeLa细胞愈合率增加,敲低ACSS2愈合率降低;营养缺乏处理上调宫颈癌细胞的侵袭能力,敲低ACSS2侵袭细胞数减少;营养缺乏处理48 h后,宫颈癌细胞中ACSS2、磷酸化的糖原合成酶激酶-3β(p-GSK-3β)以及核内β-catenin蛋白表达均有升高;敲低ACSS2后,可抑制营养缺乏诱导的Wnt/β-catenin信号活化.结论敲低ACSS2水平可以有效逆转营养缺乏诱导的宫颈癌侵袭和迁移,可能与抑制Wnt/β-catenin信号活化相关.

Objective To investigate the effect of acetyl coenzyme A synthase 2(ACSS2)on the migration and invasion induced by nutrient stress(NS)in cervical cancer cells.Methods Immunohistochemistry was used to detect the expression and distribution of ACSS2 protein in 20 pairs of cervical tumors and their adjacent normal tissues.Cervical cancer HeLa cells and the normal cervical epithelial HCvEpC cells were cultured,and serum content in culture medium was reduced from 100 to 10 miyL as NS treatment.Western blot analysis was performed to detect the expression of ACSS2,GSK-3β,p-GSK-3β,β-catenin,β-actin,E-cadherin,vimentin.Small interfering RNA(siRNA)was used to down-regulate the expression of ACSS2.Cell migration was assessed by wound healing test,and cell invasion was tested by Transwell^TM assay.Results The expression level of ACSS2 in 20 cervical tumors was significantly higher as compared with the adjacent normal tissues.The levels of ACSS2 in HeLa cells could be significantly up-regulated by NS,while no marked change was seen in HCvEpC cells.The treatment of NS promoted the epithelial mesenchymal transformation(EMT)of HeLa cells,which could be effectively reversed by siRNA-AOSS2.The scratch results showed that NS increased the healing rate of HeLa cells,which could be blocked by ACSS2 silencing.Coincidently,the number of invasive cells was elevated after NS treatment,which could be partly reversed by siRNA-ACSS2.The expression of ACSS2,p-GSK-3β and nuclear β-catenin was up-regulated in HeLa cells treated with NS for 48 hours,while siRNA-ACSS2 down-regulated their expression.Conclusion Silencing ACSS2 expression inhibits migration and invasion of cervical cancer cells induced by NS,which is related to down-regulated Wnt/β-catenin signaling pathway activity.