Department of Respiratory Medicine, Shandong Public Health Clinical Center, Jinan, Shandong 250100, China.
Department of Medical Image, Shandong Public Health Clinical Center, Jinan, Shandong 250100, China.
Pathol Res Pract. 2022 Sep;237:153941. doi: 10.1016/j.prp.2022.153941. Epub 2022 May 13.
The aberrantly expressed long non-coding RNAs (lncRNAs) are closely correlated with the malignant progression of cancer cells. In our study, we identified lncRNA MIR9-3 host gene (MIR9-3HG) as the research target and explored its roles in lung squamous cell carcinoma (LUSC). RT-qPCR was conducted to reveal that MIR9-3HG was observably overexpressed in LUSC cells. Functional assays encompassing colony formation, 5-ethynyl-2'-deoxyuridine (EdU) staining, transwell and flow cytometry assays and western blot detecting related proteins demonstrated that MIR9-3HG depletion hampered cell proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) while accelerating cell apoptosis in LUSC. Subcellular fractionation assay were performed to demonstrate that MIR9-3HG was prominently distributed in the cytoplasm of LUSC cells. Luciferase reporter, RNA-binding protein immunoprecipitation (RIP), immunofluorescence (IF), fluorescent in situ hybridization (FISH) and RNA pull down assays were implemented to confirm that MIR9-3HG modulates LIM domain kinase 1 (LIMK1) mRNA and protein levels by sequestering microRNA-138-5p (miR-138-5p) and recruiting TATA-box binding protein associated factor 15 (TAF15) protein. Taken together, our research determined that MIR9-3HG up-regulated LIMK1 mRNA and protein levels to promote LUSC carcinogenesis, which offers a novel insight into mechanisms of LUSC.
异常表达的长链非编码 RNA(lncRNA)与癌细胞的恶性进展密切相关。在我们的研究中,我们确定 lncRNA MIR9-3 宿主基因(MIR9-3HG)作为研究靶点,并探讨了其在肺鳞状细胞癌(LUSC)中的作用。实时定量 PCR 显示 MIR9-3HG 在 LUSC 细胞中明显过表达。包括集落形成、5-乙炔基-2'-脱氧尿苷(EdU)染色、Transwell 和流式细胞术以及 Western blot 检测相关蛋白的功能测定表明,MIR9-3HG 耗竭可抑制 LUSC 细胞的增殖、迁移、侵袭和上皮-间充质转化(EMT),同时加速细胞凋亡。亚细胞分馏测定表明 MIR9-3HG 主要分布在 LUSC 细胞的细胞质中。荧光素酶报告、RNA 结合蛋白免疫沉淀(RIP)、免疫荧光(IF)、荧光原位杂交(FISH)和 RNA 下拉测定证实,MIR9-3HG 通过结合 microRNA-138-5p(miR-138-5p)和募集 TATA 框结合蛋白相关因子 15(TAF15)蛋白来调节 LIM 激酶 1(LIMK1)mRNA 和蛋白水平。总之,我们的研究确定 MIR9-3HG 上调 LIMK1 mRNA 和蛋白水平以促进 LUSC 癌发生,为 LUSC 的发生机制提供了新的见解。