ATIC对乳腺癌细胞MB231迁移和增殖能力的影响

The Effect of ATIC on Migration and Proliferation of MB231 Cells

目的探讨5-氨基咪唑-4-羟酰胺核糖核酸甲酰基转移酶(ATIC)在乳腺癌中的表达情况及其对乳腺癌细胞系MB231生物学功能的影响。方法运用实时荧光定量PCR法(qPCR法)和蛋白印迹法(Western blot法)检测ATIC在9例乳腺癌患者癌组织以及癌旁组织中的表达情况;通过慢病毒干扰技术构建ATIC低表达的乳腺癌细胞系MB231并通过qPCR法和Western blot法检测ATIC的干扰效果;采用CCK-8法、平板克隆实验和EDU染色法联合检测细胞增殖情况;采用Transwell实验和划痕实验测定细胞迁移能力;Western blot法检测ATIC对上皮间质转化关键蛋白(E-cadherin和Vimentin)以及增殖相关蛋白PCNA表达的影响。结果ATIC在乳腺癌组织中的表达明显高于癌旁组织;ATIC在MB231细胞和T47D细胞中表达较高;sh-ATIC-MB231细胞系构建成功,sh-ATIC组ATIC的蛋白和mRNA水平均明显低于sh-NC组;敲低ATIC表达明显减弱乳腺癌细胞MB231的增殖和迁移能力;ATIC下调导致E-cadherin表达水平上调而Vimentin、PCNA相对表达水平下调。结论ATIC可以影响乳腺癌细胞的增殖和迁移,减缓乳腺癌的发展,有望成为乳腺癌治疗的新靶点。

Objective To explore the expression of ATIC in breast cancer tissues and discuss its function on cell proliferation and migration of MB231 cells.Methods The mRNA and protein levels of ATIC were tested by qPCR and Western blotting in 9 pairs of breast cancer tissues and normal tissues.MB231 cells were stably transfected with lentiviral shATIC vector and the expression of ATIC was detected by qPCR and Western blotting.Combined with clone formation,CCK-8 assay and EDU staining,the ability of proliferation was examined;Transwell assay and wound healing were conjointly performed to test the ability of cell migration.Western blotting was used to detect the expression levels of EMT related proteins.Results The mRNA and protein levels of ATIC were obviously increased in breast cancer than in adjacent normal tissues.sh-ATICM-B231 cells were successfully constructed,and the mRNA and protein levels of ATIC were confirmed lower in sh-ATIC group than in sh-NC group.ATIC knockdown inhibited cell proliferation,migration and EMT progress.Conclusion ATIC maybe acts an effective target in breast cancer by regulating cell proliferation,migration and EMT.