CircFLNB在骨髓源性细胞中的表达鉴定及功能分析

Expression identification and functional analysis of circFLNB in bone marrow-derived cells

摘要:

目的： 探讨环状RNA hsa_circ_0066336（circFLNB）在骨髓源性细胞中的表达，并分析其潜在功能。方法： 选择骨髓源性细胞THP-1和Jurket作为模式细胞，通过设计背对背引物获得circRNA，分析circRNA在2种模式细胞中表达的差异。采用T-A克隆手段构建质粒，进行测序、比对。用Arraystar软件预测hsa_circ_0066336的靶标miRNA。从THP-1细胞中提取靶标miRNA后反转录，再以Real-time PCR检测。结果： ①在THP-1和Jurket细胞中成功扩增出circFLNB，结果显示其表达在2种模式细胞中没有统计学差异（P=0.105）。②克隆出含有hsa_circ_0066336的质粒，测序比对后找到了反向环状剪切位点。③通过Arraystar软件预测了hsa_circ_0066336靶向的5个miRNA，并检测到靶标miRNA的表达。结论： CircFLNB是表达在骨髓源性细胞中的一个环状RNA，通过对其下游靶向miRNA功能的了解，推断circFLNB可能会参与多种疾病的发生。

Abstract:

Objective: To identify the expression of RNA hsa_circ_0066336 (circFLNB) in bone marrow-derived cells, and to analyze its potential function. Methods: Bone marrow-derived cells THP-1 and Jurket were selected as model cells. Divergent primers were designed, and differences of circRNA expression in two model cells were analyzed. T-A cloning was used to construct plasmids for comparing. Then Arraystar software was used to predict the target miRNA of hsa_circ_0066336. The target miRNA was extracted and reverse transcribed from THP-1, and then was detected by Real-time PCR. Results: ①CircFLNB was successfully amplified in THP-1 and Jurket cells, and statistical analysis showed that their expression was not significantly different in two model cells (P=0.105). ②The plasmid containing hsa_circ_0066336 was cloned, and a back-splice junction was found after comparing two sequences.③Five target miRNAs of hsa_circ_0066336 were predicted by Arraystar software and corresponding expressions were detected. Conclusion: CircFLNB is a circular RNA expressed in bone marrow derived cells and it is suggested that circFLNB may involve in the development of various diseases by comprehending its downstream miRNA function.