组织蛋白酶B在SAHA诱导乳腺癌MCF-7细胞凋亡过程中的调控作用

The regulation of Cathepsin B on cell apoptosis induced by SAHA in breast cancer cell line MCF-7

目的阐明组织蛋白酶B(cathepsin B,Cat B)在SAHA诱导的乳腺癌雌激素受体阳性细胞系MCF-7细胞凋亡中的调控作用。方法采用MTT法检测SAHA对乳腺癌MCF-7细胞生长状况的影响;应用ELISA方法测定SAHA作用于MCF-7细胞后相关蛋白表达变化的情况;通过Bio Station^(IM)活细胞工作站实时收集各种处理因素对乳腺癌MCF-7细胞增殖干预的形态学影响;并通过自动细胞分析仪Muse Cell Analyzer分析SAHA对MCF-7细胞活力和细胞凋亡的影响。结果 SAHA能明显抑制乳腺癌MCF-7细胞的增殖,其最佳作用浓度为10μmol·L^(- 1),最佳作用时间为24 h。ELISA结果表明,SAHA能诱导乳腺癌MCF-7细胞中Cat B的表达。实时活细胞工作站成像实验从形态学上证明,Cat B抑制剂Cystatin C和SAHA联合应用可有效阻止SAHA对MCF-7细胞生长的抑制作用。细胞学实验结果表明,SAHA能使MCF-7细胞活力下降,细胞凋亡发生率明显增高;然而经过Cystatin C处理后的MCF-7细胞活力增加,细胞凋亡发生率下降。结论 Cat B在SAHA诱导乳腺癌雌激素受体阳性细胞MCF-7凋亡过程中具有重要的调控作用。

To clarify the regulation role of ca-thepsin B （ Cat B ） in cell proliferation and apoptosisinduced by SAHA in ER-positive breast cancer cellline MCF-7. Methods MTT was used to screen theoptimal concentration and treatment time of SAHA .The expression levels of related proteins were determinedby ELISA,and the morphological changes wereobserved through time-lapse live cell imaging acquisition.Cell viability and apoptosis assay in MCF-7 cellswere assessed by Muse Cell Analyzer with SAHA and/or Cystatin C treatment. Results MTT assay showedthat the anti-tumor efficacy of SAHA was significant.The optimal concentration and treatment time were 10μmol·L^-1 and 24 h respectively. ELISA assayshowed that SAHA could induce expression of Cat B inMCF-7 cells. Real-time live-cell imaging experimentsdemonstrated that the combination treatment of CystatinC and SAHA significantly resumed the inhibitory effectcaused by SAHA alone. Cytology test showed that SAHA alone obviously depressed the cell viability and inducedapoptosis. However, the effect was reversedwith the combination of Cystatin C . Conclusion Cat Bplays an important role in apoptosis induced by SAH Ain ER＋ breast cancer cells MCF-7.