SAHA-CTSV轴通过诱导过度自噬抑制乳腺癌MCF-7细胞的生长

目的研究SAHA-CTSV轴通过诱导过度自噬抑制乳腺癌MCF-7细胞生长的作用和机制。方法以人乳腺癌MCF-7细胞为研究目标,Muse细胞分析仪检测SAHA对细胞活力的作用;Real-time PCR和Western blot检测SAHA对CTSV的mRNA及蛋白表达的影响,同时检测利用siRNA干扰技术敲除CTSV后的细胞CTSV表达水平;细胞免疫荧光法检测SAHA对LC3II分子的作用;Real-time PCR和Western blot检测SAHA诱导自噬过程中相关ATG分子和LC3的mRNA和蛋白的变化;Bafilomycin A1抑制自噬,随后利用Western blot和Muse细胞分析仪分别检测p62蛋白水平和SAHA作用后的细胞活力。结果SAHA可以抑制MCF-7细胞的增殖,并促进CTSV的表达。CTSV-siRNA转染细胞后,CTSV的mRNA和蛋白水平均显著性降低。SAHA增加了LC3免疫荧光点的分布,与CTSV-siRNA共同作用可恢复由CTSV-siRNA敲减引起的细胞自噬减弱。SAHA在增强ATG4C和Beclin1表达的同时,LC3B也随之升高而mTOR则下降。抑制CTSV表达后也同时逆转了SAHA的效应。Bafilomycin A1阻断自噬后,被SAHA抑制的p62蛋白的表达和细胞活力显著增强。结论SAHA-CTSV轴通过诱导过度自噬抑制乳腺癌MCF-7细胞的生长。

SAHA-CTSV induced excessive autophagy and inhibited growth of breast cancer MCF-7 cells

Aim To investigate the mechanism of SAHA-CTSV inducing autophagy and inhibiting the growth of breast cancer MCF-7 cells.Methods Muse cell analyzer was used to screen the optional treatment doses of drug for cells,and real-time PCR and Western blot were to detect the mRNA and protein expressions of CTSV.Then,specific siRNA was transfected in cells for silencing the CTSV function.Immunofluorescent assay was used to detect the distribution of LC3II,and related ATG molecule expressions were assessed.Bafilomycin A1 was used to inhibit autophagy flux,then p62 protein expression and cell viability were detected.Results 5μmol·L-1 SAHA inhibited the proliferation of MCF-7 cells accompanied with high expression of CTSV.CTSV-siRNA transfected MCF-7 cells significantly reduced the function of CTSV.With SAHA treatment,the cells increased the distribution of LC3.While knock-off the function of CTSV by siRNA,the autophagy effect was recovered.Finally,SAHA enhanced the expression of ATG4C and Beclin1,with the increase of LC3B and the decrease of mTOR.The addition of CTSV-siRNA reversed the effect of SAHA.After Bafilomycin A1 blocked autophagy,the p62 protein expression and cell viability inhibited by SAHA increased significantly.Conclusion SAHA–CTSV induces excessive autophagy and inhibits the growth of breast cancer MCF-7 cells.