Downregulation of HSP27 by isoindole-derived pyrrolidines suppressing multidrug resistance (MDR) and inducing apoptosis in MCF-7 and DLD-1 cell lines

Abstract

In most cancer treatments, major problem arises from the prevention of cell death (apoptosis suppression) with the development of drug resistance. Anticancer agents that ensure elimination of drug resistance and drug-resistant cells to apoptosis, are among the main targets. Here, we evaluated a series of synthesized N-phenyl maleimide substituents in tetracyclic compounds as anticancer drug candidate. We selected compounds may lead to death and eliminate drug resistance in breast and colon cells. In MCF-7 and DLD-1 cell lines; multidrug resistance genes (ABCB1, ABCC3, ABCC10, ABCC11 and ABCG2), apoptosis mechanism genes (BAX, BCL-2, p53, PARP and CASP3), heat shock genes (HSP27, HSP40, HSP60, HSP70 and HSP90α) and endoplasmic reticulum (ER) chaperone genes (GRP78 and GRP94) mRNA levels were determined by qPCR method. Amounts of proteins of apoptosis and signalling pathways were measured by human apoptosis antibody array. The compounds have been shown to have downregulation on multidrug resistance genes other than ABCC3. It was found that all compounds in MCF-7 and DLD-1 cells showed significant increase in p53, BAX and CASP3 gene expressions. Also, the compounds have the potential to reduce gene expression of heat shock genes (HSPs). While the compounds have been determined to increase protein expression in BAD, BAX, BID, BIM, Caspase-3, Caspase-7, Caspase-8, Cytochrome-C, Fas, TNF, TRAIL, p27, p38 and p53; decrease protein expression in AKT, BCL-2, ERK1/2, HSP27, HSP60, IGFs, JNK, NFKB, PARP, TAK1, Survivin in MCF-7 and DLD-1 cells. The compounds stand out with their inhibition of HSP27 in DLD-1 cells and their inhibition with HSP27 and NFkB in MCF-7 cells. Overall, it has been shown that these compounds increase intrinsic and extrinsic proapoptotic proteins, decrease antiapoptotic proteins, decrease HSPs and some growth factors, and they may serve as potential anticarcinogenic molecules.