不同表达模式的 HAX-1 对人星形胶质细胞增殖和凋亡的影响。

Effect of different expression patterns of HAX-1 on the proliferation and apoptosis of human astrocyte.

机构信息

Department of Orthopaedics, Affiliated Nantong Traditional Chinese Medical Hospital of Nantong University, China.

Department of Orthopaedics, Nantong Hospital of Traditional Chinese Medicine, China.

出版信息

Adv Clin Exp Med. 2022 Jun;31(6):689-699. doi: 10.17219/acem/146583.

DOI:10.17219/acem/146583

PMID:35302298

Abstract

BACKGROUND

Spinal cord injury (SCI), a serious damage of the central nervous system, has become an extremely important issue that threatens the health of people worldwide. The proliferation of astrocytes plays an important role in the repair of SCI, which has typical two-sided effects. The HS1-associated protein X-1 (HAX-1), plays an important role in the physiological and pathological processes of cell apoptosis, proliferation, migration, and invasion. However, the specific role and mechanism of HAX-1 in human astrocyte HA1800 are still unclear.

OBJECTIVES

To explore the effect of HAX-1 on the proliferation and apoptosis of HA1800 cells and preliminarily explore its possible underlying mechanism.

MATERIAL AND METHODS

The HA1800 cell lines with highand low-expression levels of HAX-1 were established using lentiviral vector pcDNA3.1. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot were employed to determine the expression of HAX-1 after transfection. Cell viability and proliferation ability were estimated using MTT and 5-Ethynyl-2'deoxyuridine (EdU) assay. The effects of HAX-1 on the HA1800 cell cycle and apoptosis were determined using flow cytometry. The BCL-2/BAX ratio and the expression of Ki67 and c-Myc in the transfected cells were detected using qRT-PCR. The Gene Expression Profiling Interactive Analysis (GEPIA) database was used to determine the relationships of HAX-1, BAX and BCL-2.

RESULTS

The HA1800 cell lines with high and low expression of HAX-1 were obtained. The MTT, EdU and flow cytometry showed that elevated HAX-1 could inhibit the proliferation, reduce the viability and promote the apoptosis of HA1800 cells. The qRT-PCR showed that the mRNA levels of Ki67, c-Myc and the BCL-2/BAX ratio were significantly decreased in the HAX-1 high-expression group, but increased in the HAX-1 low-expression group. The results from the GEPIA database showed that HAX-1 was positively correlated with BAX and BCL-2 in the spinal cord.

CONCLUSIONS

The HAX-1 may influence the biological behavior of human HA1800 cells due to the progression of cell cycle and apoptosis associated with BCL-2/BAX.

摘要

背景

脊髓损伤（SCI）是一种严重的中枢神经系统损伤，已成为威胁全球人类健康的极其重要的问题。星形胶质细胞的增殖在 SCI 的修复中起着重要作用，具有典型的两面性。HS1 相关蛋白 X-1（HAX-1）在细胞凋亡、增殖、迁移和侵袭的生理和病理过程中发挥重要作用。然而，HAX-1 在人星形胶质细胞 HA1800 中的具体作用和机制尚不清楚。

目的

探讨 HAX-1 对 HA1800 细胞增殖和凋亡的影响，并初步探讨其可能的潜在机制。

材料和方法

利用慢病毒载体 pcDNA3.1 建立 HAX-1 高表达和低表达的 HA1800 细胞系。转染后采用定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法（western blot）检测 HAX-1 的表达。MTT 和 5-乙炔基-2′-脱氧尿苷（EdU）检测法评估细胞活力和增殖能力。流式细胞术检测 HAX-1 对 HA1800 细胞周期和凋亡的影响。qRT-PCR 检测转染细胞中 BCL-2/BAX 比值以及 Ki67 和 c-Myc 的表达。利用基因表达谱分析交互分析（GEPIA）数据库确定 HAX-1、BAX 和 BCL-2 之间的关系。

结果

获得 HAX-1 高表达和低表达的 HA1800 细胞系。MTT、EdU 和流式细胞术显示，高表达 HAX-1 可抑制 HA1800 细胞的增殖，降低细胞活力并促进其凋亡。qRT-PCR 显示，HAX-1 高表达组 Ki67、c-Myc 和 BCL-2/BAX 比值的 mRNA 水平明显降低，而 HAX-1 低表达组则升高。GEPIA 数据库的结果表明，HAX-1 在脊髓中与 BAX 和 BCL-2 呈正相关。