环状 RNA circCSPP1 通过抑制 miR-200c 的成熟来促进人骨肉瘤细胞的增殖并增加葡萄糖代谢。

Circle RNA circCSPP1 promotes human osteosarcoma cell proliferation and increases glucose metabolism by suppressing miR-200c maturation.

机构信息

Experiment Center of Basic Medical Sciences of Kunming Medical University, Kunming City, P. R. China.

Department of Orthopedics, 36657The First Affiliated Hospital of Kunming Medical University, Kunming City, P. R. China.

出版信息

Hum Exp Toxicol. 2022 Jan-Dec;41:9603271221097364. doi: 10.1177/09603271221097364.

DOI:10.1177/09603271221097364

PMID:35713481

Abstract

INTRODUCTION

MiR-200c plays a central role in glucose metabolism in cancer cells. However, its upstream regulators in this process are unknown. CircRNA CSPP1 (circCSPP1) was predicted to bind to premature miR-200c, an oncogenic miRNA. Therefore, we explored their interaction in osteosarcoma (OS).

METHODS

Differential circCSPP1 and miR-200c expression in OS was analyzed using RT-qPCR. Glucose metabolism was analyzed by glucose uptake assay. Subcellular circCSPP1 location in OS cells was detected using cellular fractionation assay. The direct interaction between circCSPP1 and miR-200c was explored using RNA-RNA pull-down assay. The role of circCSPP1 in miR-200c maturation was investigated by analyzing both mature and premature miR-200c levels in OS cells with circCSPP1 overexpression.

RESULTS

CircCSPP1 and premature miR-200c levels were increased while mature miR-200c level was decreased in OS. CircCSPP1 was detected in both the nuclear and cytoplasm fractions of OS cells. CircCSPP1 directly interacted with premature miR-200c. CircCSPP1 overexpression increased premature miR-200c level, glucose uptake, and cell proliferation, but decreased mature miR-200c level. MiR-200c overexpression suppressed the role of circCSPP1 in OS cells.

CONCLUSIONS

CircCSPP1 promotes OS cell proliferation and increases glucose metabolism by suppressing miR-200c maturation.

摘要

简介

miR-200c 在癌细胞的葡萄糖代谢中发挥核心作用。然而，其在该过程中的上游调节因子尚不清楚。环状 RNA CSPP1（circCSPP1）被预测与致癌 miRNA 前体 miR-200c 结合。因此，我们在骨肉瘤（OS）中探索了它们的相互作用。

方法

使用 RT-qPCR 分析 OS 中差异表达的 circCSPP1 和 miR-200c。通过葡萄糖摄取测定法分析葡萄糖代谢。使用细胞分馏测定法检测 OS 细胞中 circCSPP1 的亚细胞定位。使用 RNA-RNA 下拉测定法探索 circCSPP1 和 miR-200c 之间的直接相互作用。通过分析 OS 细胞中 circCSPP1 过表达时成熟和前体 miR-200c 的水平，研究 circCSPP1 在 miR-200c 成熟中的作用。

结果

OS 中 circCSPP1 和前体 miR-200c 水平升高，而成熟 miR-200c 水平降低。circCSPP1 在 OS 细胞的核和细胞质部分均有检测到。circCSPP1 与前体 miR-200c 直接相互作用。circCSPP1 过表达增加前体 miR-200c 水平、葡萄糖摄取和细胞增殖，但降低成熟 miR-200c 水平。miR-200c 过表达抑制了 circCSPP1 在 OS 细胞中的作用。