Effects and mechanism of proteasome inhibitor YSY01-A alone or in combination with cisplatin against A549 cells in vitro

化合物YSY01-A是一种新型蛋白酶体抑制剂,前期研究已初步证实其具有显著的抗肿瘤作用,但是该化合物对人非小细胞肺癌细胞的作用及相关机制研究尚不明确。本实验旨在评价化合物YSY01-A单用及其与顺铂联用时,对非小细胞肺癌细胞A549的体外作用,并探讨可能的分子机制。研究表明,化合物YSY01-A对A549细胞具有显著的增殖抑制作用,且与顺铂联用后显示出较强的协同作用。分子机制研究表明,化合物YSY01-A能使PI3K表达下降,Akt的磷酸化水平降低,显著增加通路负性调节蛋白PTEN的表达。与顺铂联用后,化合物YSY01-A能显著提高细胞内的铂蓄积量,并上调CTRl蛋白的表达。综上所述,化合物Y...YSY01-A, as a novel proteasome inhibitor, has shown remarkable proliferation inhibitory effect on certain types of tumor cells. However, few studies have reported its effect on non-small cell lung cancer (NSCLC), and its underlying mechanism remains unknown. In our present study, we aimed to figure out the inhibitory effects as well as the mechanism of proteasome inhibitor YSY01-A against A549 cells both individually and in combination with cisplatin. A549 cell proliferation inhibition was assessed by SRB assay. Its related protein expression levels were determined by western blot assay. Moreover, the change of intracellular cisplatin accumulation was examined by ICP-MS assay. The results suggested that YSY01-A significantly (P<O.OOl) inhibited the proliferation of A549 cells (IC_(50) was 36.2 nM for 72 h) in a concentration-dependent and time-dependent manner. Compared with the negative control group, YSY01-A (60 nM, 48 h) down-regulated PI3K/Akt pathway in A549 cells by increasing the expression level ofPTEN (P<0.01),and decreasing the expression level of PI3K (P<0.001) and p-Akt/Akt (P<0.001). When combined with cisplatin, YSY01-A of different concentrations (5,10,20 nM) could significantly increase the inhibition effects on A549 cells compared with the cisplatin alone treatment, showing a synergistic effect. At the same time, YSY01-A could remarkably block the cisplatin-induced down-regulation of hCTRl in a concentration-dependent manner and increase cisplatin uptake from 2.01 to 2.47 fold (P<0.001). In conclusion, compound YSY01-A could significantly inhibit proliferation of NSCLC A549 cells, showing a strong synergistic effect when combined with cisplatin. Down-regulation of PI3K/Akt pathway might be the mechanism of inhibitory effect of YSY01-A, and the combination with cisplatin might increase the expression of CTR1 and intracellular cisplatin accumulation.中国科技核心期刊(ISTIC)中国科学引文数据库(CSCD)09607-6162