Ceballos María Paula, Decándido Giulia, Quiroga Ariel Darío, Comanzo Carla Gabriela, Livore Verónica Inés, Lorenzetti Florencia, Lambertucci Flavia, Chazarreta-Cifre Lorena, Banchio Claudia, Alvarez María de Luján, Mottino Aldo Domingo, Carrillo María Cristina
Instituto de Fisiología Experimental (IFISE), Facultad de Ciencias Bioquímicas y Farmacéuticas, CONICET, UNR, Suipacha 570, 2000, Rosario, Argentina.
Instituto de Fisiología Experimental (IFISE), Facultad de Ciencias Bioquímicas y Farmacéuticas, CONICET, UNR, Suipacha 570, 2000, Rosario, Argentina; Área Morfología, Facultad de Ciencias Bioquímicas y Farmacéuticas, UNR, Suipacha 570, 2000, Rosario, Argentina.
Toxicol Lett. 2018 Jun 1;289:63-74. doi: 10.1016/j.toxlet.2018.03.011. Epub 2018 Mar 12.
Sirtuins (SIRTs) 1 and 2 deacetylases are overexpressed in hepatocellular carcinoma (HCC) and are associated with tumoral progression and multidrug resistance (MDR). In this study we analyzed whether SIRTs 1 and 2 activities blockage was able to affect cellular survival and migration and to modulate p53 and FoxO1 acetylation in HepG2 and Huh7 cells. Moreover, we analyzed ABC transporters P-glycoprotein (P-gp) and multidrug resistance-associated protein 3 (MRP3) expression. We used cambinol and EX-527 as SIRTs inhibitors. Both drugs reduced cellular viability, number of colonies and cellular migration and augmented apoptosis. In 3D cultures, SIRTs inhibitors diminished spheroid growth and viability. 3D culture was less sensitive to drugs than 2D culture. The levels of acetylated p53 and FoxO1 increased after treatments. Drugs induced a decrease in ABC transporters mRNA and protein levels in HepG2 cells; however, only EX-527 was able to reduce MRP3 mRNA and protein levels in Huh7 cells. This is the first work demonstrating the regulation of MRP3 by SIRTs. In conclusion, both drugs decreased HCC cells survival and migration, suggesting SIRTs 1 and 2 activities blockage could be beneficial during HCC therapy. Downregulation of the expression of P-gp and MRP3 supports the potential application of SIRTs 1 and 2 inhibitions in combination with conventional chemotherapy.
沉默调节蛋白（SIRTs）1和2去乙酰化酶在肝细胞癌（HCC）中过表达，并与肿瘤进展和多药耐药性（MDR）相关。在本研究中，我们分析了SIRTs 1和2活性的阻断是否能够影响细胞存活和迁移，并调节HepG2和Huh7细胞中p53和FoxO1的乙酰化。此外，我们分析了ABC转运蛋白P-糖蛋白（P-gp）和多药耐药相关蛋白3（MRP3）的表达。我们使用坎比诺和EX-527作为SIRTs抑制剂。两种药物均降低了细胞活力、集落数量和细胞迁移，并增加了细胞凋亡。在三维培养中，SIRTs抑制剂减少了球体的生长和活力。三维培养比二维培养对药物的敏感性更低。处理后，乙酰化p53和FoxO1的水平升高。药物诱导HepG2细胞中ABC转运蛋白的mRNA和蛋白质水平降低；然而，只有EX-527能够降低Huh7细胞中MRP3的mRNA和蛋白质水平。这是第一项证明SIRTs对MRP3有调节作用的研究。总之，两种药物均降低了HCC细胞的存活和迁移，表明阻断SIRTs 1和2的活性在HCC治疗中可能有益。P-gp和MRP3表达的下调支持了SIRTs 1和2抑制与传统化疗联合应用的潜在可能性。
