Wild-type IDH1 Knockout Leads to G0/G1 Arrest, Impairs Cancer Cell Proliferation, Altering Glycolysis, and the TCA Cycle in Colon Cancer
Tarih
12/01/2023Üst veri
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The isocitrate dehydrogenase (IDH), which participates in the TCA cycle, is an
important key enzyme in regulating cell metabolism. The effect of the metabolic
IDH enzyme on cancer pathogenesis has recently been shown in different types of
cancer. However, the role of wild-type (wt) IDH1 in the development of colon cancer
is still unknown. Our study investigated the role of the IDH1 enzyme in key
hallmarks of colon cancer using various methods such as wound healing, cell cycle,
colony formation ability, invasion, and apoptosis analysis. Furthermore, cell metabolism
was investigated by pyruvate analysis, dinitrosalicylic acid, and HPLC methods.
In addition, CRISPR/Cas9 tool was utilized to knockout the IDH1 gene in colon
adenocarcinoma cells (SW620). Further studies were performed in two isogenic
IDH1 KO clones. Our findings in both clones suggest that IDH1 KO results in G0/
G1 arrest, and reduces proliferation by approximately twofold compared to IDH1
WT cells. In addition, the invasion, migration, and colony formation abilities of
IDH1 KO clones were significantly decreased accompanied by significant morphological
changes. In the context of metabolism, intracellular glucose, pyruvate, αKG,
and malate levels were decreased, while the intracellular citrate level was increased
in IDH1 KO clones as compared to IDH1 WT cells. Our results reveal that wt IDH1
knockout leads to a decrease in the aggressive features of colon cancer cells. In conclusion,
we reported that wt IDH1 has an effective role in colon cancer progression
and could be a potential therapeutic target.