Abstract 1563: Anoikis associated metabolic modulation by UCA1 in CRC

Abstract

Abstract Colorectal Cancer (CRC) is the second most lethal cancer in estimated deaths for 2023 according to the American Cancer Society. This is partially due to the low survival rate once CRC is diagnosed at a distant location. This warrants early diagnostic markers and an understanding of the mechanisms underlying the metastatic process, anoikis resistance being one of them. Our laboratory analysis of the TCGA database comprising 519 human CRC tissues shows lncRNA UCA1 overexpressed in tumors compared to normal, and higher expression of lncRNA UCA1 is responsible for the progression, metastasis, and poor survival of CRC patients. In our anchorage-independent growth model (anoikis resistant), using isogenic (oncogenic and metastatic) cell lines, we observed a very high expression of lncRNA UCA1 along with GluT1 with increased cell survival. Cancer cells, however, appear to rapidly increase their glucose uptake and directly ferment it into lactate, even in the presence of oxygen and functional mitochondria. UCA1 has been found to play an essential role in the development of metastasis and dysregulation of glycolysis in various cancers. To verify if lncRNA UCA1 modulates anoikis resistance and glucose metabolism, we generated SW480+UCA1 overexpression (OE) and SW620+shUCA1 knockdown (KD) stable cell lines using lentiviral transduction. The OE GFP-expressing cell lines were further sorted, and grown in puromycin. The KD cell lines were selected with puromycin for the generation of stable cell lines. The stable cell lines were characterized through different phenotypic assays. UCA1 OE cell lines have a higher glucose uptake and the UCA1 KD cell lines have a lower glucose uptake. The lactate production is high in the KD and low in OE cell lines. The OE and KD cell lines along with the control cell lines were subjected to an anoikis model, and glucose and lactate analysis were performed accordingly. RT-PCR and western blot analysis were performed for the target genes responsible for cell survival, stemness, and glucose metabolism. We are analyzing the Extracellular Acidification Rate (ECAR), Oxidative Consumption Rate (OCR), ATP production rate the rate and potential max rate of glycolysis, and the mitochondrial stress of the recombinant cell lines using the “Agilent Seahorse XF”. The role of lncRNA UCA1 in anoikis resistance leading to metastasis and modulating glucose metabolism will provide new insights into the pathophysiology of CRC metastasis. This information will be beneficial for developing more unique strategies for the treatment of this metastatic disease. Citation Format: Sophia Leslie, Kyle Doxtater, Samantha Lopez, Bilal Hafeez, Tamer Oraby, Timothy Loy, Manish K. Tripathi. Anoikis associated metabolic modulation by UCA1 in CRC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1563.