Forkhead box O1 targeting replication factor C subunit 2 expression promotes glioma temozolomide resistance and survival.
Forkhead box O1 targeting replication factor C subunit 2 expression promotes glioma temozolomide resistance and survival.
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叉头盒O1靶向复制因子C亚基2表达促进神经胶质瘤替莫唑胺耐药和存活
DOI：
10.21037/atm-21-1523
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发表时间：
2021-04
影响因子：
--
通讯作者：
Xiao S
中科院分区：
文献类型：
--
作者：
Qiu X;Tan G;Wen H;Lian L;Xiao S
背景胶质瘤中替莫唑胺(TMZ)耐药的其他机制尚不清楚。本研究的目的是确定胶质瘤中涉及叉头盒O1(FoxO1)和复制子C2(Rfc2)的另一种DNA修复机制。方法用TMZ作用于人脑胶质瘤细胞株U87R。采用增殖率细胞计数试剂盒8(CCK8)，免疫印迹法检测细胞上皮-间充质转化(EMT)相关标志物。用热图和散点图分析FoxO1与Rfc2的相关性，用实时定量逆转录聚合酶链式反应(qRT-PCR)和Western印迹法检测FoxO1对Rfc2表达的影响。通过荧光素酶报告基因检测分析FoxO1对Rfc2表达的调节作用。通过短发夹状RNA(ShRNA)实现FoxO1/Rfc2基因敲除，CCK8法和集落形成实验检测FoxO1/Rfc2基因敲除对细胞增殖的影响，流式细胞仪和免疫印迹法检测FoxO1/Rfc2细胞凋亡。结果建立了TMZ耐药胶质瘤细胞株U87R。U87R细胞中FoxO1和Rfc2蛋白表达显著增加。FoxO1和Rfc2在胶质瘤组织中的表达呈正相关。我们发现FoxO1通过调节Rfc2的表达参与TMZ耐药和细胞存活。此外，FoxO1通过与Rfc2的启动子结合，对Rfc2起转录激活作用。此外，FoxO1/Rfc2基因敲除抑制了U87R细胞的增殖，抑制了对TMZ的耐药性，并诱导了细胞的凋亡。结论FoxO1/Rfc2信号通路促进胶质瘤细胞增殖和TMZ耐药，提示FoxO1/Rfc2信号通路可能成为治疗TMZ耐药胶质瘤的潜在靶点。
Background Additional mechanisms of temozolomide (TMZ) resistance in gliomas remain uncertain. The aim of this study was to identify another DNA repair mechanism involving forkhead box O1 (FoxO1) and replicator C2 (RFC2) in gliomas. Methods We established glioma cells against TMZ, U87R, by exposure to TMZ. Proliferation rate Cell counting kit-8 (CCK8) was used, and epithelial-mesenchymal transition (EMT)-related markers were detected by western blot. The association between FoxO1 and RFC2 was analyzed by heat maps and scatter plot, and Real-time reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were used to detect the effect of FoxO1 on the expression of RFC2. The regulation effect of FoxO1 on RFC2 expression was analyzed by luciferase reporter gene assay. Knockdown of FoxO1/RFC2 was achieved via short hairpin RNA (shRNA), the effect of knockdown on the proliferation was determined by CCK8 assay and colony formation assay, and apoptosis was examined by flow cytometry and immunoblotting. Results The TMZ-resistant glioma cell line, U87R, was established. The FoxO1 and RFC2 proteins increased significantly in U87R. The expression of FoxO1 and RFC2 were positively related in glioma tissues. We found that FoxO1 contributes to TMZ resistance and cell survival via regulating the expression of RFC2. Moreover, FoxO1 functions as a transcriptional activator to RFC2 by binding to the promoter of RFC2. Furthermore, knockdown of FoxO1/RFC2 suppressed cell proliferation, TMZ resistance, and induced apoptosis in U87R. Conclusions The FoxO1/RFC2 signaling pathway promotes glioma cell proliferation and TMZ resistance, suggesting that the FoxO1/RFC2 pathway may be a potential target for TMZ-resistant glioma therapy.
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影响因子：
7.4
作者：
Melguizo C;Prados J;González B;Ortiz R;Concha A;Alvarez PJ;Madeddu R;Perazzoli G;Oliver JA;López R;Rodríguez-Serrano F;Aránega A
通讯作者：
Aránega A
影响因子：
3.5
作者：
Shiomi Y;Nishitani H
通讯作者：
Nishitani H
影响因子：
5.2
作者：
Okada, Masashi;Sato, Atsushi;Kitanaka, Chifumi
通讯作者：
Kitanaka, Chifumi
DOI：
10.1016/j.bbrc.2008.12.091
发表时间：
2009-02-06
影响因子：
3.1
作者：
Papait, Roberto;Magrassi, Lorenzo;Cattaneo, Elena
通讯作者：
Cattaneo, Elena
影响因子：
1.1
作者：
Davis ME
通讯作者：
Davis ME