新型Hsp90/HDAC双靶点抑制剂对乳腺癌细胞MDA-MB-231增殖、侵袭和迁移的影响及其机制

EFFECT OF A NOVEL HEAT SHOCK PROTEIN 90/HISTONE DEACETYLASE DUAL-TARGET INHIBITOR ON THE PROLIFE-RATION,INVASION,AND MIGRATION OF BREAST CANCER MDA-MB-231 CELLS AND ITS MECHANISM

目的 研究新型热休克蛋白90(Hsp90)/组蛋白去乙酰化酶(HDAC)双靶点抑制剂FXX-W-12-4对乳腺癌MDA-MB-231细胞增殖、侵袭和迁移能力的影响及其机制。方法 采用CCK-8实验检测FXX-W-12-4对MDA-MB-231细胞活力的影响,并计算其半致死浓度(IC50)作为后续实验中浓度设定的参考值。以IC50值为依据,在MDA-MB-231细胞中分别加入终浓度为0、100、200、300 nmol/L的FXX-W-12-4(分别设为A、B、C、D组),采用划痕实验、Transwell实验分别检测FXX-W-12-4对各组MDA-MB-231细胞侵袭以及迁移能力的影响。采用Western Blot实验检测各组MDA-MB-231细胞中ac-H3、Hsp70等蛋白的相对表达量。结果 CCK-8实验结果显示,随着FXX-W-12-4浓度的升高,MDA-MB-231细胞活力逐渐下降(F=2 663.00,P<0.05)。划痕实验结果显示,与A组相比,B~D组MDA-MB-231细胞在第12、24、48小时时迁移能力均明显降低(F=86.47~212.59,P<0.05)。Transwell实验结果显示,与A组相比,随着FXX-W-12-4浓度的升高,B~D组MDA-MB-231细胞的侵袭能力均受到了抑制(F=68.67,P<0.05)。Western Blot检测结果显示,与A组比较,随着FXX-W-12-4浓度的增加,B~D组MDA-MB-231细胞中ac-H3、Hsp70蛋白相对表达量显著性增高(F=645.60、1 017.00,P<0.05),而p-Akt、p-mTOR蛋白表达量显著性下降(F=480.30、14.32,P<0.05)。结论 新型Hsp90/HDAC双靶点抑制剂FXX-W-12-4可有效抑制乳腺癌MDA-MB-231细胞增殖、侵袭和迁移,其抑制作用可能是通过调节Akt/mTOR通路实现的。

Objective To investigate the effect of FXX-W-12-4,a novel heat shock protein 90(Hsp90)/histone deacetylase(HDAC)dual-target inhibitor,on the proliferation,invasion,and migration of breast cancer MDA-MB-231 cells.Methods CCK-8 assay was used to observe the effect of FXX-W-12-4 on the viability of MDA-MB-231 cells,and its half-maximal inhibitory concentration(IC 50)was calculated as the reference value for concentration in follow-up experiment.On the basis of the IC 50 value,FXX-W-12-4 with a final concentration of 0,100,200,and 300 nmol/L was added to MDA-MB-231 cells and was established as groups A,B,C,and D,respectively.Cell scratch assay and Transwell assay were used to observe the effect of FXX-W-12-4 on the invasion and migration of MDA-MB-231 cells in each group.Western blot was used to measure the relative expression levels of proteins such as ac-H3 and Hsp70 in MDA-MB-231 cells.Results CCK-8 assay showed that the viability of MDA-MB-231 cells gradually decreased with the increase in FXX-W-12-4 concentration(F=2663.00,P<0.05).Cell scratch assay showed that compared with group A,groups B,C,and D had a significant reduction in the migration ability of MDA-MB-231 cells at 12,24,and 48 h(F=86.47-212.59,P<0.05).Transwell assay showed that compared with group A,groups B,C,and D showed a significant reduction in the invasion ability of MDA-MB-231 cells with the increase in FXX-W-12-4 concentration(F=68.67,P<0.05).Western blot showed that compared with group A,groups B,C,and D showed significant increases in the relative protein expression levels of ac-H3 and Hsp70(F=645.60,1017.00,P<0.05)and significant reductions in the protein expression levels of p-Akt and p-mTOR(F=480.30,14.32,P<0.05)with the increase in FXX-W-12-4 concentration.Conclusion The novel Hsp90/HDAC dual-target inhibitor FXX-W-12-4 can effectively inhibit the proliferation,invasion,and migration of breast cancer MDA-MB-231 cells,possibly by regulating the Akt/mTOR pathway.