AATF knockdown reduces cell proliferation and promotes cell apoptosis in LPS-induced sepsis

The authors investigated the function of the apoptotic antagonistic transcription factor (AATF) in lipopolysaccharide (LPS)-induced H9C2 cell apoptosis and proliferation. After transfecting H9C2 cells with short hairpin RNA (shRNA), a shAATF-162 gene interference model was established. Cells were divided into mock, shRNA, and shNC groups and were treated with LPS. The cell counting kit-8 (CCK-8) assay and annexin V PE/7-AAD staining were used to assess cell viability and apoptosis, respectively. The protein levels of p53 upregulated modulator of apoptosis (PUMA) and phorbol-12-myristate-13-acetate-induced protein 1 (NOXA) were determined using western blotting. Interleukin 6 and tumor necrosis factor alpha (TNF-alpha) secretion was determined using enzyme-linked immunosorbent assay (ELISA) and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Results of the CCK-8 assay showed that AATF knockdown decreased proliferation and potentiated apoptosis in LPS-induced H9C2 cells compared with that in the shRNA and mock groups. The western blotting results indicated that AATF knockdown increased PUMA and NOXA protein levels in LPS-induced H9C2 cells compared to that in the shNC and mock groups. ELISA and RT-qPCR results showed that AATF knockdown significantly decreased the LPS-induced secretion of IL-6 and TNF-alpha in H9C2 cells compared to that in the shNC and mock groups. This study showed that AATF inhibition attenuated cell viability, potentiated apoptosis, and decreased the LPS-induced secretion of IL-6 and TNF-alpha in H9C2 cells.