阿帕替尼调控上皮间质转化抑制三阴性乳腺癌MDA-MB-231细胞侵袭

Apatinib inhibits invasion of triple negative breast cancer MDA-MB-231 cells by modulating epithelial-mesenchymal transition

目的探讨阿帕替尼对人三阴性乳腺癌MDA-MB-231细胞侵袭作用的影响。方法MTT法检测不同浓度的阿帕替尼(1、2、4、8、16μmol·L^-1)作用于乳腺癌MDA-MB-231细胞48h的细胞毒性,并采用Bliss法计算半数抑制浓度(IC50);Annexin V-FITC/PI流式细胞术检测阿帕替尼对MDA-MB-231细胞凋亡的影响;Transwell实验检测阿帕替尼对MDA-MB-231细胞侵袭能力的影响;Western blot检测阿帕替尼对上皮间质转化(EMT)标志性蛋白上皮钙黏素、神经钙黏素及波形蛋白表达的影响。结果1、2、4、8、16μmol·L^-1阿帕替尼可显著抑制MDA-MB-231细胞的增殖,增殖抑制率分别为(38.38±3.33)%、(53.17±4.31)%、(62.09±6.21)%、(80.97±7.50)%和(98.54±9.75)%。阿帕替尼作用48 h对MDA-MB-231细胞的IC50为1.96μmol·L^-1。与空白对照组相比,阿帕替尼可显著诱导MDA-MB-231细胞凋亡,显著抑制MDA-MB-231细胞侵袭基底膜的能力(P<0.05);阿帕替尼还可显著上调MDA-MB-231细胞上皮钙黏素的表达,下调神经钙黏素及波形蛋白的表达(P<0.05)。结论阿帕替尼可能通过调控EMT进而发挥抗三阴性乳腺癌MDA-MB-231细胞侵袭作用。

AIM To investigate the effects of apatinib on the invasion of human triple negative breast cancer MDA-MB-231 cells.METHODS MDA-MB-231 cells were treated by apatinib at the varying concentrations(1,2,4,8,16μmol·L^-1)for 48 h and the cytotoxicity were detected by MTT assay.The IC50 value of apatinib in MDA-MB-231 cells was calculated by Bliss method.Annexin V-FITC/PI staining was used to observe the effect of apatinib on apoptosis of MDA-MB-231 cells by flow cytometry.Transwell assay was used to investigate the effect of apatinib on the invasion of MDA-MB-231 cells.The effects of apatinib on the expression of epithelial-mesenchymal transition(EMT)related proteins including E-cadherin,N-cadherin and vimentin were detected by Western blot.RESULTS Apatinib(1,2,4,8,16μmol·L^-1)significantly inhibited the proliferation of MDA-MB-231 cells,and the proliferation inhibition rates were(38.38±3.33)%,(53.17±4.31)%,(62.09±6.21)%,(80.97±7.50)%and(98.54±9.75)%,respectively.The IC50 of apatinib in MDA-MB-231 cells was 1.96μmol·L^-1 for 48 h.Compared with the blank control group,apatinib significantly induced apoptosis and suppressed the invasion efficacies of MDA-MB-231 cells(P<0.05).The E-cadherin expression level was upregulated and the N-cadherin and vimentin expression levels were down-regulated in MDA-MB-231 cells by apatinib(P<0.05).CONCLUSION Apatinib could inhibit the invasion of triple negative breast cancer MDAMB-231 cells by modulating EMT.