TAF15 inhibits p53 nucleus translocation and promotes HCC cell 5-FU resistance via post-transcriptional regulation of UBE2N.

IF 3.7 3区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Jiayan Fang, Mengqi Zou, Mei Yang, Yejia Cui, Rong Pu, Yufeng Yang
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引用次数: 0

Abstract

Chemotherapy resistance is an important factor responsible for the low 5-year survival rate of hepatocellular carcinoma (HCC) patients. Ubiquitin-conjugating enzyme E2N (UBE2N) is a cancer-associated ubiquitin-conjugating enzyme that is expressed in HCC tissues, and its high expression is associated with a poor prognosis. This study explored the role played by UBE2N in development of 5-fluorouracil (5-FU) resistance in HCC cells. Three HCC cell lines (HepG2 [p53 wild type], Huh7 [p53 point mutant type], Hep3B [p53 non-expression type]), and one normal liver cell line (MIHA) were used in our present study. The IC50 value of 5-FU was determined using a cell counting kit-8 (CCK-8) assay. Cell viability was assessed by colony formation assays. TUNEL assays and flow cytometry were used to analyze cell apoptosis. RNA pull-down and RNA immunoprecipitation (RIP) assays were performed to confirm the binding relationship between UBE2N mRNA and TAF15 protein. Our results showed that TAF15 and UBE2N were highly expressed in HCC cells. UBE2N inhibited the translocation of p53 protein into the cell nucleus to increase 5-FU resistance, as reflected by an increased IC50 value, an increase in cell viability, and a reduction in cell apoptosis. Overexpression of p53 reduced 5-FU resistance, but that effect could be reversed by UBE2N overexpression. TAF15 protein bound to and stabilized UBE2N mRNA, thereby inhibiting p53 translocation into the nucleus and promoting 5-FU resistance in HCC cells. Collectively, our present study identified a novel mechanism by which TAF15/UBE2N regulates p53 distribution to increase 5-FU resistance. Our results also suggest potential therapeutic strategies for treating HCC.

TAF15 通过转录后调控 UBE2N 抑制 p53 核转位并促进 HCC 细胞对 5-FU 的耐受。
化疗耐药性是导致肝细胞癌(HCC)患者 5 年生存率低的一个重要因素。泛素结合酶E2N(UBE2N)是一种与癌症相关的泛素结合酶,在HCC组织中表达,它的高表达与预后不良有关。本研究探讨了 UBE2N 在 HCC 细胞对 5-氟尿嘧啶(5-FU)产生耐药性的过程中所起的作用。本研究使用了三种 HCC 细胞系(HepG2 [p53 野生型]、Huh7 [p53 点突变型]、Hep3B [p53 非表达型])和一种正常肝细胞系(MIHA)。用细胞计数试剂盒-8(CCK-8)测定 5-FU 的 IC50 值。细胞活力通过集落形成试验进行评估。TUNEL检测和流式细胞术用于分析细胞凋亡。为了证实 UBE2N mRNA 与 TAF15 蛋白之间的结合关系,进行了 RNA 拉取和 RNA 免疫沉淀(RIP)试验。结果表明,TAF15和UBE2N在HCC细胞中高表达。UBE2N 可抑制 p53 蛋白向细胞核内的转位,从而增加对 5-FU 的耐药性,具体表现为 IC50 值升高、细胞活力增加和细胞凋亡减少。过表达 p53 可降低 5-FU 抗性,但过表达 UBE2N 可逆转这种效应。TAF15 蛋白与 UBE2N mRNA 结合并使其稳定,从而抑制了 p53 转位至细胞核,促进了 HCC 细胞的 5-FU 抗性。总之,本研究发现了 TAF15/UBE2N 调节 p53 分布以增加 5-FU 抗性的新机制。我们的研究结果还为治疗 HCC 提出了潜在的治疗策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of physiology and biochemistry
Journal of physiology and biochemistry 生物-生化与分子生物学
CiteScore
6.60
自引率
0.00%
发文量
86
审稿时长
6-12 weeks
期刊介绍: The Journal of Physiology and Biochemistry publishes original research articles and reviews describing relevant new observations on molecular, biochemical and cellular mechanisms involved in human physiology. All areas of the physiology are covered. Special emphasis is placed on the integration of those levels in the whole-organism. The Journal of Physiology and Biochemistry also welcomes articles on molecular nutrition and metabolism studies, and works related to the genomic or proteomic bases of the physiological functions. Descriptive manuscripts about physiological/biochemical processes or clinical manuscripts will not be considered. The journal will not accept manuscripts testing effects of animal or plant extracts.
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