PO.MCB10.02 · 分子与细胞生物学
LncRNA UCA1调控LDHA表达以驱动Warburg代谢和结直肠癌进展
LncRNA UCA1 regulates LDHA expression to drive Warburg metabolism and colorectal cancer progression
该海报暂无可下载的资料
AACR 官方页面
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：结直肠癌（CRC）是全球第三大最常见的恶性肿瘤，也是美国第二大死亡原因。2025年，预计将诊断出154,270例新发结肠癌病例。在美国，结直肠癌是癌症相关死亡的第二大最常见原因，2025年预计死亡52,900例。尿路上皮癌相关1（UCA1）是一种长链非编码RNA（lncRNA），已被发现在CRC中失调并促进肿瘤生长。与致癌性结肠细胞不同，我们观察到lncRNA UCA1在转移性CRC细胞中显著表达。其表达还与葡萄糖代谢因子相关，包括葡萄糖转运体1（Glut1）和乳酸脱氢酶（LDHA）。LDHA对癌细胞快速产生ATP至关重要，从而支持其进展。
方法：对同基因CRC细胞（SW480和SW620），以及过表达UCA1的细胞（SW480+UCA1；OE）和UCA1敲低的细胞（SW620+shUCA1；KD）进行细胞迁移、侵袭和增殖试验。使用同基因、OE和KD细胞系检查失巢凋亡诱导。采用RT-PCR、Western blot和免疫荧光技术对YAP1、Glut1和LDHA进行RNA和蛋白分析。还评估了CRC细胞中的葡萄糖和乳糖水平。
结果：与致癌性CRC（SW480）细胞相比，lncRNA UCA1在转移性CRC（SW620）细胞中表达显著更高。在失巢凋亡条件下36小时后，UCA1表达呈指数级升高。相比致癌性（SW480细胞），UCA1水平及Glut1和YAP1在转移（SW620细胞）中也升高。在UCA1过表达的SW480细胞中，Glut1和YAP1的基因表达水平显著增加，表明UCA1在调控葡萄糖代谢中发挥作用。在Warburg效应中，癌细胞即使在有氧存在下也将丙酮酸转化为乳酸，而非将其用于TCA循环。我们发现，将丙酮酸转化为乳酸的酶LDHA的基因表达在转移性CRC细胞中显著高于致癌性CRC细胞。UCA1过表达细胞也显示LDHA表达显著增加，提示UCA1影响LDHA酶活性。LDHA与GLUT1和YAP1相关，突显了LDHA在Warburg葡萄糖代谢和CRC细胞进展中的关键作用。抑制LDHA可降低细胞内Glut1水平并限制肿瘤细胞生长。
查看英文原文 English abstract
Background : Colorectal cancer (CRC) is the third most common malignancy worldwide and the second leading cause of death in the US. In 2025, an estimated 154,270 new cases of colon cancer will be diagnosed. In the US, colorectal cancer ranks as the second most common cause of cancer-related deaths, with an expected 52,900 deaths in 2025. Urothelial cancer-associated 1 (UCA1), a long noncoding RNA (lncRNA), has been found to be dysregulated in CRC and to promote tumor growth. Unlike carcinogenic colon cells, we have observed that the lncRNA UCA1 is significantly expressed in metastatic CRC cells. Its expression is also linked to glucose metabolism factors, including glucose transporter 1 (Glut1) and lactate dehydrogenase (LDHA). LDHA is essential for the rapid production of ATP in cancer cells, thereby supporting their progression.
Methods: Cell migration, invasion, and proliferation assays were conducted on isogenic CRC cells (SW480 and SW620), as well as on cells overexpressing UCA1 (SW480+UCA1; OE) and those with UCA1 knockdown (SW620+shUCA1; KD). Anoikis induction was examined using the isogenic, OE, and KD cell lines. RT-PCR, Western blot, and immunofluorescence techniques were employed for RNA and protein analysis of YAP1, Glut1, and LDHA. Glucose and lactose levels in CRC cells were also assessed.
Results: LncRNA UCA1 showed significantly higher expression in metastatic CRC (SW620) cells compared to carcinogenic CRC (SW480) cells. After 36 hours under anoikis conditions, UCA1 expression became exponentially higher. UCA1 levels, along with Glut1 and YAP1, are also elevated in metastasis (SW620 cells) compared to carcinogenic (SW480 cells). In UCA1-overexpressing SW480 cells, the gene expression levels of Glut1 and YAP1 increased significantly, indicating that UCA1 plays a role in regulating glucose metabolism. In the Warburg effect, cancer cells convert Pyruvate into lactate even in the presence of oxygen, rather than using it for the TCA cycle. We found that the gene expression of LDHA, the enzyme converting pyruvate to lactate, was significantly higher in metastatic CRC cells compared to carcinogenic CRC cells. UCA1-overexpressing cells also showed significantly increased LDHA expression, suggesting that UCA1 influences LDHA enzyme activity. LDHA correlated with GLUT1 and YAP1, highlighting the crucial role of LDHA during Warburg glucose metabolism and CRC cell progression. Inhibiting LDHA reduces Glut1 levels within the cell and limits tumor cell growth.
利益披露 Disclosure
S. H. Shaham, None..
R. Pequeno Bracho, None..
K. Renteria, None..
M. Tripathi, None.