Cancer Biology Division, School of Biotechnology, Kalinga Institute of Industrial Technology (KIIT), Deemed to Be University, Campus-11, Patia, Bhubaneswar, 751024, Odisha, India.
Cancer Biology Division, School of Biotechnology, Kalinga Institute of Industrial Technology (KIIT), Deemed to Be University, Campus-11, Patia, Bhubaneswar, 751024, Odisha, India.
DNA Repair (Amst). 2021 Sep;105:103157. doi: 10.1016/j.dnarep.2021.103157. Epub 2021 Jun 10.
Apart from inducing catalytic inhibition of PARP-1, PARP inhibitors can also trap PARP proteins at the sites of DNA damage and forming toxic PARP-DNA complexes. These complexes obstruct the DNA repair process, resulting in cancer cell death. To study the detailed mechanism of anti-cancer action through PARP trapping, we have treated oral cancer cells (H-357) with curcumin (Cur), olaparib (Ola) and their combination (Cur + Ola). Cur + Ola treatment triggered the expressions of PARP-1 and adenomatous polyposis coli (APC) and down regulated other base excision repair (BER) proteins in the chromatin fraction but not in the nuclear fraction. Cur + Ola treatment inhibited PARylation, altered interaction of PARP-1 with representative BER proteins and arrested cells in S-phase. We have for the first time provided direct evidence and measured the cellular PARP-1 trapping potentiality of Ola in Cur pretreated H-357 cells. Unchanged cellular PARP-1 trapping, unaltered expression of BER proteins and BER activity were found in APC silenced H-357 cells, which further confirmed that the DNA damage/repair response was APC-dependent. Interestingly, complete abolishment of the chromatin remodeler 'amplified in Liver Cancer 1' (ALC1), decreased expression of Histone H3 and histone acetyltransferase (P300) was noted in chromatin of Cur + Ola treated cells. Their expressions remained unchanged in APC silenced cells. Cur + Ola also altered the interaction of ALC1 with BER proteins including APC. Thus, the present study reveals that Cur + Ola treatment increased oral cancer cell death not only through catalytic inhibition of PARP-1 but also predominantly through PARP-1 trapping and indirect inhibition of chromatin remodeling.
除了诱导 PARP-1 的催化抑制外,PARP 抑制剂还可以将 PARP 蛋白捕获在 DNA 损伤部位,并形成有毒的 PARP-DNA 复合物。这些复合物会阻碍 DNA 修复过程,导致癌细胞死亡。为了研究通过 PARP 捕获发挥抗癌作用的详细机制,我们用姜黄素(Cur)、奥拉帕利(Ola)及其组合(Cur+Ola)处理口腔癌细胞(H-357)。Cur+Ola 处理触发了 PARP-1 和腺瘤性结肠息肉病基因(APC)的表达,并下调了染色质部分而非核部分的其他碱基切除修复(BER)蛋白。Cur+Ola 处理抑制了 PAR 化,改变了 PARP-1 与代表性 BER 蛋白的相互作用,并使细胞停滞在 S 期。我们首次提供了直接证据,并测量了 Cur 预处理的 H-357 细胞中 Ola 的细胞 PARP-1 捕获潜力。在 APC 沉默的 H-357 细胞中,发现细胞 PARP-1 捕获不变,BER 蛋白表达不变,BER 活性不变,这进一步证实了 DNA 损伤/修复反应依赖于 APC。有趣的是,在 Cur+Ola 处理的细胞中,染色质重塑剂“肝癌扩增物 1”(ALC1)完全消失,组蛋白 H3 和组蛋白乙酰转移酶(P300)的表达减少。在 APC 沉默的细胞中,它们的表达保持不变。Cur+Ola 还改变了 ALC1 与包括 APC 在内的 BER 蛋白的相互作用。因此,本研究表明,Cur+Ola 处理不仅通过对 PARP-1 的催化抑制,而且主要通过 PARP-1 捕获和间接抑制染色质重塑,增加了口腔癌细胞的死亡。