Department II of General Surgery, Hanchuan People's Hospital, Hanchuan, Hubei, China.
Department of Gynaecology and Obstetrics, Hanchuan People's Hospital, Hanchuan, Hubei, China.
Histol Histopathol. 2023 Oct;38(10):1129-1143. doi: 10.14670/HH-18-544. Epub 2022 Nov 4.
Circular RNAs (circRNAs) are key molecules in the regulation of intrahepatic cholangiocarcinoma (ICC) progression. The purpose of this study was to analyze the function and underlying molecular mechanism of circ_0000284 in ICC.
Quantitative real-time PCR was used to analyze the circ_0000284, microRNA (miR)-152-3p and pyruvate dehydrogenase kinase 1 (PDK1) expression. Cell proliferation, apoptosis, invasion and migration were executed by cell counting kit 8 assay, EdU assay, flow cytometry, transwell assay and wound healing assay, respectively. All protein expression levels were examined using western blot analysis. Cell glycolysis was analyzed by detecting glucose consumption, lactate production and ATP/ADP ratios. Target relationship was estimated by dual-luciferase reporter assay. The effect of circ_0000284 on ICC tumor growth in vivo was evaluated by constructing xenograft mice model.
We detected high expression of circ_0000284 in ICC tumor tissues and cells. Downregulated circ_0000284 inhibited ICC cell proliferation, invasion, migration, glycolysis, and accelerated apoptosis. MiR-152-3p was sponged by circ_0000284, and its inhibitor revoked the effect of circ_0000284 knockdown on ICC cell progression. PDK1 was a target of miR-152-3p, and its expression was suppressed by circ_0000284 knockdown. PDK1 overexpression reversed the inhibition effect of miR-152-3p on ICC cell growth, metastasis and glycolysis. In animal experiments, circ_0000284 downregulation also inhibited ICC tumor growth.
Circ_0000284 promoted the growth, metastasis and glycolysis of ICC by miR-152-3p/PDK1 pathway, showing that circ_0000284 was a potential therapeutic target for ICC.
环状 RNA（circRNAs）是调控肝内胆管癌（ICC）进展的关键分子。本研究旨在分析 circ_0000284 在 ICC 中的功能及潜在分子机制。
采用实时定量 PCR 分析 circ_0000284、微小 RNA（miR）-152-3p 和丙酮酸脱氢酶激酶 1（PDK1）的表达。通过细胞计数试剂盒 8 检测、EdU 检测、流式细胞术、Transwell 检测和划痕愈合试验分别检测细胞增殖、凋亡、侵袭和迁移。通过 Western blot 分析检测所有蛋白表达水平。通过检测葡萄糖消耗、乳酸生成和 ATP/ADP 比值分析细胞糖酵解。通过双荧光素酶报告实验评估靶标关系。通过构建异种移植小鼠模型评估 circ_0000284 对 ICC 肿瘤生长的体内影响。
我们检测到 ICC 肿瘤组织和细胞中 circ_0000284 高表达。下调 circ_0000284 抑制 ICC 细胞增殖、侵袭、迁移、糖酵解，并加速凋亡。circ_0000284 可吸附 miR-152-3p，其抑制剂可逆转 circ_0000284 敲低对 ICC 细胞进展的影响。PDK1 是 miR-152-3p 的靶标，circ_0000284 敲低可抑制其表达。PDK1 过表达逆转了 miR-152-3p 对 ICC 细胞生长、转移和糖酵解的抑制作用。在动物实验中，circ_0000284 下调也抑制了 ICC 肿瘤的生长。
circ_0000284 通过 miR-152-3p/PDK1 通路促进 ICC 的生长、转移和糖酵解，表明 circ_0000284 可能是 ICC 的潜在治疗靶点。
