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配对盒5诱导的LINC00467上调通过触发微小RNA-4735-3p/肿瘤坏死因子α诱导蛋白3通路促进喉鳞状细胞癌进展。

Paired Box 5-Induced LINC00467 Upregulation Promotes the Progression of Laryngeal Squamous Cell Cancer by Triggering the MicroRNA-4735-3p/TNF Alpha-Induced Protein 3 Pathway.

作者信息

Li Yuan, Wu Yong, Dai Lili, Wu Honglin, Chen Chaohui, Ni Jiayu, Jin Enli, Zhou Xuehua

机构信息

Department of Otolaryngology Head and Neck Surgery, Affiliated Hospital of Hangzhou Normal University, No. 126, Wenzhou Road, Hangzhou, 310015, Zhejiang Province, China.

Department of Otorhinolaryngology, The Third People's Hospital of Deqing, Deqing, China.

出版信息

Mol Biotechnol. 2023 Apr;65(4):655-667. doi: 10.1007/s12033-022-00564-3. Epub 2022 Oct 10.

Abstract

LINC00467 was reported as an oncogenic gene in different types of human cancers. In this study, we investigated the molecular mechanisms of LINC00467 in the tumorigenesis of laryngeal squamous cell cancer (LSCC). RT-qPCR was utilized to detect the mRNA expression of genes, and western blot assay was used to determine the protein levels of TNF alpha-induced protein 3 (TNFAIP3). The cell viability was detected by CCK-8 assay. Transwell assays were conducted to determine the cell migration and invasion of LSCC cells, and the cell cycle was assessed by flow cytometry. The association between paired box 5 (PAX5), LINC00467, miR-4735-3p, and TNFAIP3 was verified using ChIP, RNA pull-down, or luciferase reporter assays. In our study, we found that LINC00467 was upregulated in LSCC, and knockdown of LINC00467 suppressed cell viability and metastasis of LSCC. Besides, LINC00467 transcription could be activated by PAX5 in LSCC. Furthermore, LINC00467 acted as competitive endogenous RNA (ceRNA) for miR-4735-3p to accelerate LSCC progression. In the meantime, TNFAIP3 was identified as a downstream gene of miR-4735-3p. Finally, TNFAIP3 overexpression could overturn the effects of miR-4735-3p mimic on LSCC cellular activities. In conclusion, our results demonstrated that PAX5-induced LINC00467 facilitated LSCC progression by inhibiting miR-4735-3p to increase TNFAIP3 expression.

摘要

LINC00467在不同类型的人类癌症中被报道为致癌基因。在本研究中,我们探究了LINC00467在喉鳞状细胞癌(LSCC)肿瘤发生中的分子机制。采用RT-qPCR检测基因的mRNA表达,并用蛋白质免疫印迹法测定肿瘤坏死因子α诱导蛋白3(TNFAIP3)的蛋白水平。通过CCK-8法检测细胞活力。进行Transwell实验以确定LSCC细胞的迁移和侵袭能力,并通过流式细胞术评估细胞周期。使用染色质免疫沉淀(ChIP)、RNA下拉或荧光素酶报告基因实验验证配对盒5(PAX5)、LINC00467、miR-4735-3p和TNFAIP3之间的关联。在我们的研究中,我们发现LINC00467在LSCC中上调,敲低LINC00467可抑制LSCC的细胞活力和转移。此外,PAX5可激活LSCC中LINC00467的转录。此外,LINC00467作为miR-4735-3p的竞争性内源性RNA(ceRNA)加速LSCC进展。同时,TNFAIP3被鉴定为miR-4735-3p的下游基因。最后,TNFAIP3过表达可逆转miR-4735-3p模拟物对LSCC细胞活性的影响。总之,我们的结果表明,PAX5诱导LINC00467通过抑制miR-4735-3p增加TNFAIP3表达来促进LSCC进展。

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