Effect of high temperature requirement protease A1 lentivirus expression vector infection on biological behavior of human vascular smooth muscle cells / 中华神经医学杂志

ABSTRACT

Objective To construct the lentivirus vectors of high temperature requirement protease A1 (HTRA 1) gene as well as it's mutant gene (1091 T＞C HTRA 1),and investigate the effect of their infection on proliferation,migration and apoptosis of human brain vascular smooth muscle cells (HBVSMC).Methods The lentivirus vectors of wild type HTRA1 and mutant HTRA1 were constructed,cultured and identified,and then,infected into HBVSMC.Empty lentivirus vectors were used as controls.CCK-8 method was used to detect the cell proliferation for a consecutive five d.Transwell assay was used to detect the cell migration,and cell apoptosis was detected by flow cytometry.Results As compared with those from the control group,cells from wild-type HTRA 1 group showed no statistically different proliferation rate (P＞0.05),while cells from mutant HTRA 1 group had significantly reduced proliferation rate since the third d (P＜0.05).Migration rate of cells from control group,wild type HTRA 1 group and mutant HTRA 1 group was 0.474±0.079,0.612±0.037,and 0.283±0.064,respectively,with significant differences (P＜0.05).Apoptosis percentage of cells from control group,wild type HTRA 1 group was 3.68％±0.23％ and 3.13％±0.07％,with significant difference (P＜0.05),while that of mutant H TRA 1 groupwas3.70％±0.20％,showingnosignificantdifferenceascomparedwiththatfromcontrolgroup (P＞0.05).Conclusion HBVSMC infected by HTRA 1 mutant gene display attenuation of proliferation and migration activity,but have no statistical change in apoptosis.