Abstract:
AIM To investigate the effects of telomerase reverse transcriptase (TERT) over-expression and RNA interference lentiviral vector on the proliferation of hypoxic pulmonary artery smooth muscle cells (PASMCs).
METHODS Plvx-IRES-ZsGreen 1-TERT over-expression vector and Plvx-shRNA 2-TERT RNA interference vector were constructed, respectively. PASMCs were cultured and divided into a normal control group, a hypoxia control group, a blank vector group, a TERT over-expression group, and a TERT RNA interference group. Cell proliferation was detected by CCK-8 method, and apoptosis and cell proliferation were detected by flow cytometry.
RESULTS Plvx-IRES-ZsGreen1-TERT over-expression vector and Plvx-shRNA2-TERT RNA interference vector were successfully constructed and sequenced. Compared with those in the normal control group, the absorbance of the hypoxic control group increased significantly, and the proportion of S phase cells increased (P < 0.05). The absorbance of the TERT over-expression group at 24 h and 48 h after hypoxia was significantly higher than that in the hypoxic control group, and G1 phase cells numbers at 48 h after hypoxia were much more than those in the hypoxic control group (P < 0.05). The absorbance value of the TERT interference group significantly decreased at 24 h and 48 h after hypoxia, the proportion of G1-phase cells was significantly increased at 48 h hypoxia, the proportion of S-phase cells significantly decreased, and the apoptosis rate significantly increased (P < 0.05).
CONCLUSION TERT RNA interference promotes apoptosis and decreases the synthesis of nucleic acid in PASMCs during hypoxia, thus inhibiting the proliferation of PASMCs.
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