PO.MCB10.02 · 分子与细胞生物学
长链非编码RNA LINC01614过表达定义甲状腺乳头状癌中一种铁死亡相关的分子特征
Long non-coding RNA LINC01614 overexpression defines a ferroptosis-associated molecular signature in papillary thyroid cancer
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摘要 Abstract
中文摘要
甲状腺乳头状癌（PTC）是最常见的内分泌恶性肿瘤，发病率每年约上升3%。PTC对育龄女性的影响尤为突出，而男性患者往往表现出更具侵袭性的疾病。尽管总生存率较高，复发和转移仍是重大的临床挑战，可在初次诊断后持续数十年。因此，鉴定可靠的预后生物标志物和可操作的治疗靶点至关重要。长链非编码RNA（lncRNA）是一类在基因表达中具有多种作用的调控分子，呈现组织特异性和癌症特异性表达模式，是生物标志物发现的有前景的候选者。对我们PTC与正常甲状腺基因组数据库的生物信息学分析发现，lncRNA LINC01614转录本在PTC中显著上调12倍。因此，我们正在研究LINC01614作为潜在的PTC生物标志物和/或治疗靶点。发现LINC01614在多种甲状腺癌细胞系中表达上调。选取PTC细胞系K1（上调约2倍；BRAFV600E；男性）用于体外研究。采用CRISPR干扰（CRISPRi）在K1细胞中转录性抑制LINC01614。K1中LINC01614敲低导致迁移减少（约20%）、克隆形成能力降低（约34%）和增殖减少（约45%），表明该lncRNA在支持恶性表型中发挥作用。对CRISPRi敲低细胞系与CRISPRi对照的RNA测序分析显示，关键铁死亡相关基因SLC7A11和SLC3A2表达降低。这两个基因均编码xCT系统（胱氨酸/谷氨酸逆向转运体），调控胱氨酸的摄取——胱氨酸是谷胱甘肽（GSH）合成的关键前体。GSH是抵御脂质过氧化的关键抗氧化防御，进而防止铁死亡。因此，这些发现提示LINC01614参与了铁死亡相关机制。与此一致，对细胞内总铁水平的定量显示，我们的LINC01614敲低细胞中铁水平显著升高，表明铁死亡诱导增强。Western blot分析还显示铁死亡相关蛋白如GPX4水平降低，GPX4是一种重要的抗氧化酶，可还原脂质过氧化物以防止铁死亡。目前正在进行研究以阐明LINC01614在铁死亡通路中的作用机制。这些发现提示LINC01614通过调节铁死亡相关过程促进PTC的发生，凸显其作为生物标志物和/或治疗靶点的潜在价值。
查看英文原文 English abstract
Papillary thyroid cancer (PTC) is the most common endocrine malignancy, with incidence rising by approximately 3% each year. PTC disproportionately affects females of reproductive age, while male patients often exhibit more aggressive disease. Although overall survival is high, recurrence and metastasis remain significant clinical challenges that can persist for decades after initial diagnosis. Identifying reliable prognostic biomarkers and actionable therapeutic targets is therefore essential. Long non-coding RNAs (lncRNAs), a class of regulatory molecules with diverse roles in gene expression, display tissue- and cancer-specific expression patterns, making them promising candidates for biomarker discovery. Bioinformatic analysis of our PTC vs. normal thyroid genomic repository identified the lncRNA LINC01614 transcript as significantly 12-fold upregulated in PTC. Thus, we are studying LINC01614 as a potential PTC biomarker and/or therapeutic target. LINC01614 expression was found to be upregulated in multiple thyroid cancer cell lines. The PTC cell line K1 (~2 fold upregulated; BRAFV600E; male), was selected for in vitro study. CRISPR interference (CRISPRi) was used to transcriptionally repress LINC01614 in K1 cells. LINC01614 knockdown in K1 resulted in decreased migration (~20%), clonogenicity (~34%) and proliferation (~45%), indicating a role of this lncRNA in supporting malignant phenotypes. RNA sequencing analysis of a CRISPRi knockdown cell line vs. a CRISPRi control demonstrated reduced expression of key ferroptosis-related genes- SLC7A11 and SLC3A2. Both genes encode for the xCT system (cystine/glutamate reverse transporter) that regulates the uptake of cystine - a key precursor for glutathione (GSH) synthesis. GSH is a critical antioxidant defense against lipid peroxidation, further preventing ferroptosis. Thus, these findings suggested LINC01614 involvement in a ferroptosis-related mechanism. Consistent with this, total cell iron levels were quantified and found to be significantly higher in our LINC01614 knockdowns, signifying increased ferroptosis induction. Western blot analyses also showed decreased levels of ferroptosis-associated proteins such as GPX4, an important antioxidant enzyme that reduces lipid peroxidation to prevent ferroptosis. Current studies are ongoing to identify a LINC01614 mechanism of action within the ferroptosis pathway. These findings suggest that LINC01614 contributes to PTC pathogenesis through modulation of ferroptosis-related processes, highlighting its potential utility as a biomarker and/or therapeutic target.
利益披露 Disclosure
D. Quaranto, None..
N. R. DeSouza, None..
S. Dadafarin, None..
A. Moscatello, None..
H. K. Islam, None..
C. Iacob, None..
R. K. Tiwari, None..
J. Geliebter, None.