NHC Key Laboratory of Hormones and Development(Tianjn Medical University), Tianjin Key Laboratory of Metabolic Diseases, Tianjn Medical University Chu Hsien-I Memorial Hospital & Tianjin Institute of Endocrinology, Tianjin 300134, China.
Tianjin Key Laboratory of Medical Epigenetics, 2011 Collaborative Innovation Center of Tianjin for Medical, Epigenetics, Department of Biochemistry and Molecular Biology, Tianjin Medical University, Tianjin, China.
Life Sci. 2019 Sep 1;232:116602. doi: 10.1016/j.lfs.2019.116602. Epub 2019 Jun 26.
Blood glucose dysregulation is an adverse factor in the prognosis of hepatocellular carcinoma (HCC). Endoplasmic reticulum (ER) is thought to be crucial component in the development of cancer and diabetes. This study aimed to investigate the mechanisms of poor outcomes in HCC patients with diabetes.
ER protein 29 (ERp29) was predicted by proteomics, immunohistochemistry, Western blot, Cell Counting Kit-8 (CCK-8) and cell scratch test were used to identify the expression and biological effects of ERp29 under high glucose (HG) in HCC cells. Bioinformatics found a competing endogenous RNAs (ceRNAs) regulatory network between microRNA-483-3p (miR-483-3p) and Long noncoding RNA (LncRNA MEG3), the above methods also were used to identify their expression, biological effects and their roles of HG on regulation of REp29 in HCC cells, Dual-luciferase reporter assay was carried out to study the interaction of ERp29 with miR-483-3p and miR-483-3p with MEG3.
HG upregulated miR-483-3p expression in HCC cells and miR-483-3p overexpression suppressed ERp29 expression and also increased HCC cell proliferation and migration. Furthermore, we found that MEG3 was decreased in HCC cells incubated in medium with high glucose and knockdown of MEG3 downregulated ERp29 expression. Bioinformatics analysis found that MEG3 mediated its protective effects via binding to miR-483-3p.
Overall, our study established a novel regulatory network of LncRNA MEG3/miR483-3p/ERp29 in HCC which may be helpful in better understanding the effect of high glucose on poor prognosis of HCC and in exploring new diagnostic and therapeutic tools for managing HCC in patients with diabetes.
血糖调节紊乱是肝细胞癌（HCC）预后不良的一个因素。内质网（ER）被认为是癌症和糖尿病发展的关键组成部分。本研究旨在探讨糖尿病合并 HCC 患者预后不良的机制。
通过蛋白质组学预测内质网蛋白 29（ERp29），用免疫组织化学、Western blot、细胞计数试剂盒-8（CCK-8）和细胞划痕试验检测高糖（HG）条件下 HCC 细胞中 ERp29 的表达及其生物学效应。生物信息学发现 microRNA-483-3p（miR-483-3p）和长非编码 RNA（LncRNA MEG3）之间存在竞争内源性 RNA（ceRNA）调控网络，上述方法也用于鉴定其表达、生物学效应及其在 HG 对 HCC 细胞中 REp29 调节中的作用，双荧光素酶报告基因实验研究 ERp29 与 miR-483-3p 以及 miR-483-3p 与 MEG3 的相互作用。
HG 上调 HCC 细胞中 miR-483-3p 的表达，miR-483-3p 过表达抑制 ERp29 表达，并增加 HCC 细胞增殖和迁移。此外，我们发现高糖培养的 HCC 细胞中 MEG3 减少，MEG3 敲低下调 ERp29 表达。生物信息学分析发现 MEG3 通过与 miR-483-3p 结合发挥其保护作用。
总之，本研究建立了 HCC 中 LncRNA MEG3/miR483-3p/ERp29 的新调控网络，有助于更好地理解高糖对 HCC 预后不良的影响，并探索用于管理糖尿病合并 HCC 患者的新的诊断和治疗工具。
