Institute of Cancer Stem Cell, Dalian Medical University, Dalian, China.
State Key Laboratory of Molecular Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Cell Prolif. 2020 Apr;53(4):e12750. doi: 10.1111/cpr.12750. Epub 2020 Mar 4.
LOC100133669 is a lncRNA whose function during tumorigenesis remains unclear now. Thus, we aimed to explore its clinical significance and function in oesophageal squamous cell carcinoma (ESCC).
ISH was used to detect LOC100133669 expression in ESCC tissues. The full-length LOC100133669 was identified by using RACE assay. Subcellular distribution of LOC100133669 was examined by nuclear/cytoplasmic RNA fractionation and qPCR. The role of LOC100133669 in ESCC cell growth was determined by colony formation, MTT and flow cytometry experiments in vitro, as well as xenograft tumour experiment in vivo. RNA pull-down assay was performed to find LOC100133669-interacted protein, which was further examined by RIP, IP, Western blot and rescue experiments.
LOC100133669 was upregulated in ESCC tissues compared with adjacent non-tumour tissues. High LOC100133669 expression was associated with poor prognosis of patients with ESCC. We defined LOC100133669 to be 831 nt in length and mainly localized in the cytoplasm of ESCC cells. Knockdown of LOC100133669 inhibited ESCC cell proliferation and cell cycle progression, while overexpression of LOC100133669 showed the opposite effects. Furthermore, LOC100133669 could bind to Tim50 and upregulated its protein level through inhibiting ubiquitination. Overexpression of Tim50 in part abolished the LOC100133669 depletion-caused inhibitory effect on ESCC cell proliferation.
LOC100133669 plays an oncogenic role in ESCC and may serve as a promising diagnostic marker and therapeutic target for ESCC patients.
LOC100133669 是一种 lncRNA,其在肿瘤发生过程中的功能尚不清楚。因此,我们旨在探讨其在食管鳞状细胞癌(ESCC)中的临床意义和功能。
采用原位杂交法检测 ESCC 组织中 LOC100133669 的表达。采用 RACE 法鉴定全长 LOC100133669。通过核/细胞质 RNA 分离和 qPCR 检测 LOC100133669 的亚细胞分布。通过体外集落形成、MTT 和流式细胞术实验以及体内异种移植肿瘤实验,确定 LOC100133669 在 ESCC 细胞生长中的作用。通过 RNA 下拉实验寻找 LOC100133669 相互作用蛋白,进一步通过 RIP、IP、Western blot 和挽救实验进行检测。
与相邻非肿瘤组织相比,LOC100133669 在 ESCC 组织中上调。LOC100133669 高表达与 ESCC 患者的预后不良相关。我们定义 LOC100133669 的长度为 831nt,主要定位于 ESCC 细胞的细胞质中。LOC100133669 的敲低抑制了 ESCC 细胞的增殖和细胞周期进程,而 LOC100133669 的过表达则表现出相反的效果。此外,LOC100133669 可以与 Tim50 结合,并通过抑制泛素化来上调其蛋白水平。Tim50 的过表达在一定程度上消除了 LOC100133669 耗竭对 ESCC 细胞增殖的抑制作用。
LOC100133669 在 ESCC 中发挥致癌作用,可能成为 ESCC 患者有前途的诊断标志物和治疗靶点。