Department of Gynaecology, ZIBO Central Hospital, No. 54 Gongqingtuan West Road, Zhangdian District, Zibo, 255036, Shandong, China.
Cell Biochem Biophys. 2020 Dec;78(4):447-453. doi: 10.1007/s12013-020-00939-w. Epub 2020 Sep 8.
The human antigen R (HuR) could play an essential role in stabilizing the mRNAs of many tumor-associated genes. Little research is performed to investigate the relevant mechanism mediated by HuR to promote the progress of ovarian cancer. The Cancer Genome Atlas (TCGA) dataset was retrieved to calculate the correlation between HuR and translocase of inner mitochondrial membrane 44 (TIMM44) expression. HuR expression plasmid, TIMM44 expression plasmid, siRNA HuR, and TIMM44 siRNAs were further transfected into A2780 and SKOV3 cells. The 3'UTR of TIMM44 fragment was cloned into the back of Renilla luciferase in the pSicheck2 dual fluorescent reporter to indicate the interaction between HuR and TIMM44. Cell count and MTT assay were performed to assay the proliferation ability of A2780 and SKOV3 cells. High-level HuR expression in 56 ovarian cancer patients recruited in Zibo Central Hospital was positively correlated with metastasis status and poor prognosis revealed by Kaplan-Meier analysis. Both HuR and TIMM44 can promote the proliferation of SKOV3 and A2780 cells. A high correlation of HuR and TIMM44 expression was testified in the TCGA data. Luciferase reporter assay confirmed that HuR could bind to TIMM44 to maintain the mRNA stability. TIMM44 siRNA administration inhibited the proliferation of SKOV3 cells, which could not be rescued. All of these indicate that the main function of HuR on ovarian cancer proliferation is mediated by TIMM44 through mRNA stability regulation, and HuR/TIMM44 complex can be used as a target to inhibit the proliferation of ovarian cancer cells.
人类抗原 R（HuR）可以在稳定许多肿瘤相关基因的 mRNA 方面发挥重要作用。很少有研究探讨 HuR 介导的促进卵巢癌进展的相关机制。从癌症基因组图谱（TCGA）数据集中计算 HuR 与内膜转位酶 44（TIMM44）表达之间的相关性。进一步将 HuR 表达质粒、TIMM44 表达质粒、HuR siRNA 和 TIMM44 siRNAs 转染到 A2780 和 SKOV3 细胞中。将 TIMM44 3'UTR 片段克隆到 Renilla 荧光素酶的 pSicheck2 双荧光报告基因的后端，以指示 HuR 和 TIMM44 之间的相互作用。细胞计数和 MTT 测定用于测定 A2780 和 SKOV3 细胞的增殖能力。淄博市中心医院招募的 56 名卵巢癌患者中高水平的 HuR 表达与 Kaplan-Meier 分析显示的转移状态和不良预后呈正相关。TCGA 数据证实 HuR 和 TIMM44 均可促进 SKOV3 和 A2780 细胞的增殖。HuR 和 TIMM44 表达的高相关性在 TCGA 数据中得到证实。荧光素酶报告基因测定证实 HuR 可以与 TIMM44 结合以维持 mRNA 稳定性。TIMM44 siRNA 给药抑制了 SKOV3 细胞的增殖，这种抑制作用不能被挽救。所有这些都表明，HuR 对卵巢癌增殖的主要功能是通过调节 mRNA 稳定性由 TIMM44 介导的，HuR/TIMM44 复合物可作为抑制卵巢癌细胞增殖的靶点。
