Pathophysiological roles of autophagy and aldo-keto reductases in development of doxorubicin resistance in gastrointestinal cancer cells
Pathophysiological roles of autophagy and aldo-keto reductases in development of doxorubicin resistance in gastrointestinal cancer cells
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DOI：
10.1016/j.cbi.2019.108839
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发表时间：
2019-12-01
影响因子：
5.1
通讯作者：
Ikari, Akira
中科院分区：
文献类型：
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作者：
Matsunaga, Toshiyuki;Kawabata, Saori;Ikari, Akira
在这里，我们表明，孵育三个人胃肠道癌细胞系（HCT 15，LoVo和MKN 45）与阿霉素（DOX）引起自噬通过促进活性氧（ROS）的产生。用DOX处理HCT 15细胞导致Beclin 1上调，Bcl 2下调，AMPK和JNK激活，Akt失活，所有这些都通过用抗氧化剂N-乙酰-L-半胱氨酸预处理恢复。这些数据表明，DOX引起的所有自噬相关的改变都是由ROS产生引起的。在DOX抗性癌细胞中，DOX引起的自噬程度比亲本细胞温和，并且DOX处理几乎没有激活ROS依赖性凋亡信号[4-羟基-2-壬烯醛（HNE）的形成，细胞色素-c释放到细胞质中，以及JNK和半胱天冬酶-3的激活]，推断细胞抗氧化能力与DOX诱导的自噬之间存在负相关性。对亲代和DOX抗性细胞中醛酮还原酶（AKR）表达水平的监测显示，随着获得DOX抗性，AKR 1B 10和/或AKR 1C 3上调。在这些细胞中敲低和抑制AKR 1B 10或AKR 1C 3增强了DOX引起的自噬。DOX-还原酶活性的测量和HNE-敏感性测定也表明AKR 1B 10（通过高HNE-还原酶活性）和AKR 1C 3（通过低HNE-还原酶和DOX-还原酶活性）都参与DOX抗性的发展。自噬抑制剂和两种AKR的组合克服了胃肠道癌细胞对DOX或顺式二氨二氯铂的耐药性发展的DOX耐药性和交叉耐药性。因此，与抑制剂的伴随治疗可能是有效的辅助治疗，用于提高胃肠道癌细胞的DOX敏感性。
Here, we show that incubation of three human gastrointestinal cancer cell lines (HCT15, LoVo and MKN45) with doxorubicin (DOX) provokes autophagy through facilitating production of reactive oxygen species (ROS). HCT15 cell treatment with DOX resulted in up-regulation of Beclin1, down-regulation of Bcl2, activation of AMPK and JNK, and Akt inactivation, all of which were restored by pretreating with an antioxidant N-acetyl-L-cysteine. These data suggest that all the autophagy-related alterations evoked by DOX result from the ROS production. In the DOX-resistant cancer cells, degree of autophagy elicited by DOX was milder than the parental cells, and DOX treatment hardly activated the ROS-dependent apoptotic signals [formation of 4-hydroxy-2-nonenal (HNE), cytochrome-c release into cytosol, and activation of JNK and caspase-3], inferring an inverse correlation between cellular antioxidant capacity and autophagy induction by DOX. Monitoring of expression levels of aldoketo reductases (AKRs) in the parental and DOX-resistant cells revealed an up-regulation of AKR1B10 and/or AKR1C3 with acquiring the DOX resistance. Knockdown and inhibition of AKR1B10 or AKR1C3 in these cells enhanced DOX-elicited autophagy. Measurement of DOX-reductase activity and HNE-sensitivity assay also suggested that both AKR1B10 (via high HNE-reductase activity) and AKR1C3 (via low HNE-reductase and DOX-reductase activities) are involved in the development of DOX resistance. Combination of inhibitors of autophagy and the two AKRs overcame DOX resistance and cross-resistance of gastrointestinal cancer cells with resistance development to DOX or cis-diamminedichloroplatinum. Therefore, concomitant treatment with the inhibitors may be effective as an adjuvant therapy for elevating DOX sensitivity of gastrointestinal cancer cells.