Mitochondrial DNA mutations are involved in the acquisition of cisplatin resistance in human lung cancer A549 cells
Mitochondrial DNA mutations are involved in the acquisition of cisplatin resistance in human lung cancer A549 cells
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DOI：
10.3892/or.2021.8243
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发表时间：
2022-02-01
期刊：
影响因子：
4.2
通讯作者：
Rikitake, Yoshiyuki
中科院分区：
文献类型：
--
作者：
Horibe, Sayo;Ishikawa, Kaori;Rikitake, Yoshiyuki
顺铂（CDDP）作为单一疗法以及与免疫疗法的联合疗法在治疗各种癌症中的疗效已得到证实。然而，获得性 CDDP 耐药性是成功治疗的主要障碍。在本研究中，使用 ACR20 细胞检查了获得性 CDDP 耐药的机制，ACR20 细胞是源自 A549 肺癌细胞的 CDDP 耐药细胞。 CDDP 通过结合核 DNA 并产生活性氧 (ROS) 来诱导细胞毒性。与我们的预期相反，未用 CDDP 处理的 ACR20 细胞中 ROS 水平升高。 ROS抑制剂预处理增强了ACR20细胞对CDDP的敏感性，并阻止核因子（NF）-kB信号传导的激活和凋亡蛋白抑制剂（IAP）的上调。值得注意的是，对线粒体耗氧率和线粒体超氧化物水平的评估揭示了 ACR20 细胞中线粒体功能的恶化。线粒体 DNA PCR-RFLP 分析显示 ACR20 细胞中有四种不同百分比水平的突变。此外，在与 ACR20 细胞线粒体的细胞质杂合体中，内在 ROS 水平升高，IAP 表达增加，复合物 I 活性和对 CDDP 的敏感性降低。三维结构数据分析表明，突变（ND2 F40L）可能影响质子易位途径，从而影响线粒体复合物I活性。总之，这些发现表明，线粒体 DNA 突变导致内在 ROS 水平升高，从而通过激活 NF-κ B 信号传导和诱导 ACR20 细胞中的 IAP 表达来降低对 CDDP 的敏感性。这些发现表明，新发现的线粒体 DNA 突变可能导致癌症获得性顺铂耐药。
The efficacy of cisplatin (CDDP) has been demonstrated in the treatment of various cancers as monotherapy and combination therapy with immunotherapy. However, acquired CDDP resistance is a major obstacle to successful treatment. In the present study, the mechanisms underlying acquired CDDP resistance were examined using ACR20 cells, which are CDDP-resistant cells derived from A549 lung cancer cells. CDDP induces cytotoxicity by binding nuclear DNA and generating reactive oxygen species (ROS). Contrary to our expectation, ROS levels were elevated in ACR20 cells not treated with CDDP. Pretreatment with an ROS inhibitor enhanced the sensitivity of ACR20 cells to CDDP and prevented the activation of nuclear factor (NF)-kB signaling and upregulation of inhibitor of apoptosis proteins (IAPs). Notably, evaluation of the mitochondrial oxygen consumption rate and mitochondrial superoxide levels revealed a deterioration of mitochondrial function in ACR20 cells. Mitochondrial DNA PCR-RFLP analysis revealed four mutations with varying percentage levels in ACR20 cells. In addition, in cytoplasmic hybrids with mitochondria from ACR20 cells, intrinsic ROS levels were elevated, expression of IAPs was increased, and complex I activity and sensitivity to CDDP were decreased. Analysis of three-dimensional structure data indicated that a mutation (ND2 F40L) may impact the proton translocation pathway, thereby affecting mitochondrial complex I activity. Together, these findings suggest that intrinsic ROS levels were elevated by mitochondrial DNA mutations, which decreased the sensitivity to CDDP via activation of NF-kappa B signaling and induction of IAP expression in ACR20 cells. These findings indicate that newly identified mutations in mitochondrial DNA may lead to acquired cisplatin resistance in cancer.