RRM2 protects against ferroptosis and is a tumor biomarker for liver cancer.
RRM2 protects against ferroptosis and is a tumor biomarker for liver cancer.
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RRM2 可防止铁死亡，并且是肝癌的肿瘤生物标志物
DOI：
10.1186/s12935-020-01689-8
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发表时间：
2020-12-07
影响因子：
5.8
通讯作者：
Wang J
中科院分区：
文献类型：
--
作者：
Yang Y;Lin J;Guo S;Xue X;Wang Y;Qiu S;Cui J;Ma L;Zhang X;Wang J
背景铁凋亡是由脂质过氧化物引发的细胞死亡过程，谷胱甘肽（GSH）合成受到抑制导致铁凋亡。肝癌进展与铁凋亡抑制密切相关。然而，谷胱甘肽合成抑制肝癌细胞的潜在铁凋亡和铁凋亡相关的肝癌生物标志物是否有一个有前途的诊断价值的机制仍然未知。MethodsRibonucleotide reductase regulatory subunit M2（RRM2）的水平进行了测量，采用酶联免疫吸附试验（ELISA），定量RT-PCR（qPCR），免疫印迹（IB）和免疫化学（IHC）。分别通过CellTiter-Glo发光细胞活力测定和SYTOX绿色染色，然后通过流式细胞术测量细胞活力和细胞死亡。使用指定的试剂盒测量甲硫氨酸。用免疫荧光法（IF）、免疫共沉淀法（co-IP）和近端连接法（PLA）检测了谷胱苷肽合成酶（GSS）与RRM2的相互作用。采用受试者工作特征曲线下面积（AUC-ROC）分析诊断价值。结果RRM2在肝癌中特异性升高，并通过GSS刺激GSH合成抑制铁凋亡。从机制上讲，RRM2在ThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeThreeTh而在亚铁胁迫下，RRM2在T33发生去磷酸化，从而促进了RRM2与GSS的相互作用。这导致RRM2和GSS易位到蛋白酶体中进行同时降解。在临床上，血清RRM2与血清甲胎蛋白（AFP）、癌胚抗原（CEA）、丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、碱性磷酸酶（ALP）、γ-谷氨酰转肽酶（γ-GT）、白蛋白（ALB）和总胆红素显著相关。RRM2与AFP联合检测的AUC-ROC为0.947，敏感性为88.7%，特异性为97.0%，较RRM2或AFP单独检测具有更好的诊断性能。血清RRM2可作为一种生物标志物，用于评估铁凋亡被抑制的程度，并提高肝癌的诊断效率。
BackgroundFerroptosis is the process of cell death triggered by lipid peroxides, and inhibition of glutathione (GSH) synthesis leads to ferroptosis. Liver cancer progression is closely linked to ferroptosis suppression. However, the mechanism by which inhibition of GSH synthesis suppresses potential ferroptosis of liver cancer cells and whether ferroptosis-related liver cancer biomarkers have a promising diagnostic value remain unknown.MethodsRibonucleotide reductase regulatory subunit M2 (RRM2) levels were measured using an enzyme linked immunosorbent assay (ELISA), quantitative RT-PCR (qPCR), immunoblotting (IB) and immunochemistry (IHC). Cell viability and cell death were measured by a CellTiter-Glo luminescent cell viability assay and staining with SYTOX Green followed by flow cytometry, respectively. Metabolites were measured using the indicated kits. The Interaction between glutathione synthetase (GSS) and RRM2 was measured using immunofluorescence (IF), co-immunoprecipitation (co-IP) and the proximal ligation assay (PLA). The diagnostic value was analyzed using the area under the receiver operating characteristic curve (AUC-ROC). Bioinformatics analysis was performed using the indicated database.ResultsRRM2 showed specifically elevated levels in liver cancer and inhibited ferroptosis by stimulating GSH synthesis via GSS. Mechanistically, phosphorylation of RRM2 at the Threonine 33 residue (T33) was maintained at normal levels to block the RRM2–GSS interaction and therefore protected RRM2 and GSS from further proteasome degradation. However, under ferroptotic stress, RRM2 was dephosphorylated at T33, thus the RRM2–GSS interaction was promoted. This resulted in the translocation of RRM2 and GSS to the proteasome for simultaneous degradation. Clinically, serum RRM2 was significantly associated with serum alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma glutamyl transpeptidase (γ-GT), albumin (ALB) and total bilirubin. The AUC-ROC for the combination of RRM2 with AFP was 0.947, with a sensitivity of 88.7% and a specificity of 97.0%, which indicates better diagnostic performance compared to either RRM2 or AFP alone.ConclusionRRM2 exerts an anti-ferroptotic role in liver cancer cells by sustaining GSH synthesis. Serum RRM2 will be useful as a biomarker to evaluate the degree to which ferroptosis is suppressed and improve diagnostic efficiency for liver cancer.
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影响因子：
14.8
作者：
Kagan VE;Mao G;Qu F;Angeli JP;Doll S;Croix CS;Dar HH;Liu B;Tyurin VA;Ritov VB;Kapralov AA;Amoscato AA;Jiang J;Anthonymuthu T;Mohammadyani D;Yang Q;Proneth B;Klein-Seetharaman J;Watkins S;Bahar I;Greenberger J;Mallampalli RK;Stockwell BR;Tyurina YY;Conrad M;Bayır H
通讯作者：
Bayır H
影响因子：
64.5
作者：
Dixon SJ;Lemberg KM;Lamprecht MR;Skouta R;Zaitsev EM;Gleason CE;Patel DN;Bauer AJ;Cantley AM;Yang WS;Morrison B 3rd;Stockwell BR
通讯作者：
Stockwell BR
影响因子：
11.2
作者：
Furuta, Eiji;Okuda, Hiroshi;Kobayashi, Aya;Watabe, Kounosuke
通讯作者：
Watabe, Kounosuke
影响因子：
16.6
作者：
Gandhi, Minakshi;Gross, Matthias;Diederichs, Sven
通讯作者：
Diederichs, Sven
DOI：
10.1073/pnas.1415518111
发表时间：
2014-11-25
影响因子：
11.1
作者：
Linkermann, Andreas;Skouta, Rachid;Krautwald, Stefan
通讯作者：
Krautwald, Stefan