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

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

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.

Keywords: HA1800; HAX-1; apoptosis; proliferation.

MeSH terms

  • Adaptor Proteins, Signal Transducing* / genetics
  • Adaptor Proteins, Signal Transducing* / metabolism
  • Apoptosis* / genetics
  • Apoptosis* / physiology
  • Astrocytes* / metabolism
  • Cell Line
  • Cell Proliferation* / genetics
  • Cell Proliferation* / physiology
  • Gene Expression
  • Gene Expression Profiling
  • Humans
  • Ki-67 Antigen
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Spinal Cord Injuries / genetics
  • Spinal Cord Injuries / metabolism
  • bcl-2-Associated X Protein / genetics
  • bcl-2-Associated X Protein / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • HAX1 protein, human
  • Ki-67 Antigen
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-2-Associated X Protein