Title: Inhibit C reactive protein induced apoptosis by silencing NF-κB/p65 gene in human airway epithelial cells
Abstract: Objective
To investigate the influence of C reactive protein (CRP) induced apoptosis through sliencing nuclear factor-kappa B p65 (NF-κB)/p65 gene by RNA interference in human airway epithelial cells 9HTE.
Methods
9HTE were cultured in vitro and chemically synthesized small interference RNA (siRNA) directed against human NF-κB/p65 was transfected into 9HTE by using cationic liposome LipofectamineTM2000 as the transfecting agent.Then after incubation with CRP for 24 h, the level of NF-κB/p65, Bcl-2 and Bax mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR), the activation of NF-κB/p65 was tested by immunocytochemical stain, the level of Bcl-2 and Bax protein were tested by Western blot, the rate of apoptosis was tested by flow cytometry.
Results
Compared with normal group, chemically synthesized small interfering RNA inhibited expression of NF-κB/p65mRNA and the activation of NF-κB/p65 in 9HTE.The expression of Bcl-2 mRNA and protein decreased significantly (P<0.05), the expression of Bax mRNA and protein increased significantly (P<0.05), the rate of apoptosis decreased significantly (P<0.05) compared with CRP and scramble siRNA groups.
Conclusions
Silencing NF-κB/p65 gene by using siRNA interfere could effectively inhibit CRP-mediated apoptosis in human airway epithelial cells.It is indicate that the activation of NF-κB has a role of pro-apoptosis in airway epithelial cells, which one of the mechanisms may be through regulating the expression of Bcl-2 and Bax to induce apoptosis.
Key words:
RNA interference; C-reactive protein; Nuclear factor-kappa B; Apoptosis; Epithelial cell
Publication Year: 2019
Publication Date: 2019-01-05
Language: en
Type: article
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