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Abstract：
We aimed to explore the function and underlying mechanism of highly upregulated in liver cancer (HULC; an long noncoding RNAs) in hepatocellular carcinoma (HCC) and chemosensitivity of oxaliplatin (Oxa). The expression of HULC, miR-383-5p, and vesicle-associated membrane protein-2 (VAMP2) was detected by quantitative real-time polymerase chain reaction. Western blot assay was applied for measuring the protein expression of cyclinD1, cleaved-caspase-3, light Chain 3 I/II, p62, and VAMP2. Cell viability and Oxa IC50 value were determined by Cell Counting Kit-8 assay. A colony formation assay was conducted to evaluate colony formation ability. Cell apoptosis was assessed by flow cytometry. The interaction between miR-383-5p and HULC or VAMP2 was predicted by bioinformatics analysis and verified by dual-luciferase reporter assay and RNA immunoprecipitation assay. The mice xenograft model was established to investigate the roles of HULC in vivo. HULC and VAMP2 were overexpressed whereas miR-383-5p was lowly expressed in HCC tissues. HULC overexpression promoted the progression of HCC cells and inhibited chemosensitivity of Oxa by increasing cell proliferation and protective autophagy and inhibiting apoptosis, whereas HULC silence presented opposite effects. Moreover, miR-383-5p was a direct target of HULC and miR-383-5p reversed the effects of HULC on the progression of HCC cells and chemosensitivity of Oxa. Besides, HULC acted as a molecular sponge of miR-383-5p to regulate VAMP2 expression. HULC promoted the progression of HCC and inhibited Oxa sensitivity by regulating miR-383-5p/VAMP2 axis, elucidating a novel regulatory mechanism for chemosensitivity of Oxa and providing a potential lncRNA-targeted therapy for HCC.
摘要：
我们旨在探讨肝癌（HULC;长链非编码RNA）在肝细胞癌（HCC）和奥沙利铂（Oxa）的化学敏感性中高度上调的功能和潜在机制。HULC的表达，miR-383-5p,通过定量实时聚合酶链反应检测囊泡相关膜蛋白2（VAMP2）。Westernblot检测cyclinD1、cleaved-caspase-3、轻链3I/II的蛋白表达,p62和VAMP2。通过细胞计数试剂盒-8测定确定细胞活力和OxaIC50值。进行集落形成测定以评价集落形成能力。通过流式细胞术评估细胞凋亡。通过生物信息学分析预测miR-383-5p与HULC或VAMP2之间的相互作用，并通过双荧光素酶报告实验和RNA免疫沉淀实验进行验证。建立小鼠异种移植模型以研究HULC在体内的作用。HULC和VAMP2过表达，而miR-383-5p在HCC组织中低表达。HULC过表达通过增加细胞增殖、保护性自噬和抑制凋亡,促进HCC细胞的进展,抑制Oxa的化疗敏感性,而HULC沉默呈现相反的效果。此外，miR-383-5p是HULC的直接靶标，miR-383-5p逆转了HULC对HCC细胞进展和Oxa化学敏感性的影响。此外,HULC作为miR-383-5p的分子海绵调节VAMP2的表达。HULC通过调节miR-383-5p/VAMP2轴促进HCC进展并抑制Oxa敏感性,阐明Oxa化学敏感性的新调节机制，并为HCC提供潜在的lncRNA靶向治疗。
