ABSTRACT

Curcumin is an antiproliferative phytochemical extracted from Curcuma longa L and which has been studied in preclinical drug screening using cell monolayers and animal models. However, several limitations of these culture systems may be overcome by performing screening with three-dimensional (3-D) cell culture. The aim of this study was to investigate the effects of curcumin on cytotoxicity and genotoxicity as well as spheroid growth using cervical adenocarcinoma HeLa cell spheroids by performing RT-PCR mRNA expression of genes involved in cell death (CASP3, CASP8, CASP9, PARP1, BBC3, BIRC5, BCL2, TNF), autophagy (BECN1, SQSTM1), cell cycle regulation (TP53, C-MYC, NF-kB, CDKN1A, m-TOR, TRAF-2), DNA damage repair (H2AFX, GADD45A, GADD45G), oxidative stress (GPX1), reticulum stress (EIF2AK3, ERN1), and invasion (MMP1, MMP9) was investigated. Curcumin was cytotoxic in a concentration-dependent manner. Curcumin-treated spheroids exhibited lower proliferative recovery and cell proliferation attenuation, as observed in the clonogenic assay. Further, no marked genotoxicity was detected. Curcumin-treated spheroids displayed reduced expression of BECN1 (2.9×), CASP9 (2.1×), and PARP1 (2.1×) mRNA. PARP1 inhibition suggested disruption of essential pathways of proliferation maintenance. Downregulated expression of CASP9 mRNA and unchanged expression of CASP3/8 mRNA suggested caspase-independent cell death, whereas downregulated expression of BECN1 mRNA indicated autophagic disruption. Therefore, curcumin exhibits the potential for drug development with antiproliferative activity to be considered for use in cancers.

摘要

姜黄素是一种从姜黄中提取的抗增殖植物化学物质，已使用单层细胞和动物模型进行临床前药物筛选研究。然而，通过三维 (3-D) 细胞培养进行筛选可以克服这些培养系统的一些局限性。本研究的目的是通过对宫颈腺癌 HeLa 细胞球体进行 RT-PCR 检测参与细胞死亡的基因 ( CASP3 , CASP8 , CASP9 , PARP1 , BBC3 ) mRNA 表达，探讨姜黄素对细胞毒性和基因毒性以及球体生长的影响。、BIRC5、BCL2、TNF）、自噬（BECN1、SQSTM1）、细胞周期调节（TP53、C-MYC、NF-kB、CDKN1A、m-TOR、TRAF-2）、DNA损伤修复（H2AFX、GADD45A、GADD45G）研究了氧化应激（GPX1）、网状应激（EIF2AK3、ERN1）和侵袭（MMP1、MMP9 ）。姜黄素具有浓度依赖性的细胞毒性。正如在克隆形成实验中观察到的，姜黄素处理的球体表现出较低的增殖恢复和细胞增殖减弱。此外，没有检测到明显的遗传毒性。姜黄素处理的球体显示BECN1 (2.9×)、CASP9 (2.1×) 和PARP1 (2.1×) mRNA 的表达降低。PARP1抑制表明增殖维持的重要途径被破坏。CASP9 mRNA表达下调和CASP3/8 mRNA表达不变表明细胞死亡不依赖于 caspase，而BECN1 mRNA 表达下调表明自噬破坏。因此，姜黄素表现出开发具有抗增殖活性的药物的潜力，可考虑用于癌症。