H3K9ac modification was involved in doxorubicin induced apoptosis by regulating Pik3ca transcription in H9C2 cells

Aim:We evaluated the effect of H3K9ac modification on the transcriptional level of Pik3ca and the apoptosis induction effects of Pik3ca in the PI3K/AKT pathway in rat H9C2 cells.

Main methods:H9C2 cells were cultured with 1uM doxorubicin for 8 h. The acetylation level of histone H3K9 in the transcriptional initiation area of Pik3ca was determined by CHIP-seq. The enrichment of mRNA fragment of Pik3ca transcription initiation region by H3K9ac antibody was detected by CHIP-qPCR, and the expression of Pik3ca was detected by real-time Polymerase Chain Reaction(rt-PCR) and western blot. The transcription efficiency of Pik3ca siRNA was detected by immunofluorescence and western blot. Cell Counting Kit8(CCK8) was used to detect the cell activity and flow cytometry was used to detect the apoptosis rate. Western blot was applied to assess the protein expression level of PI3K, P-AKT, AKT, Bcl2, BAX and cleaved-caspase3 in H9C2 cells.

Key findings:In doxorubicin-inducedH9C2 cells, the acetylation levels of histone H3K9 in the Pik3ca transcriptional initiation region significantly increased and promoted the transcription of Pik3ca. After knockdown of Pik3ca, the PI3K/AKT signaling pathway was inhibited, and the protein expression of Bcl2 was increased, the protein expression of BAX and cleaved-caspase3 were decreased. PI3K/AKT pathway activator partially reversed the effect of silencing Pik3ca.

Significance:In summary, the elevated H3K9ac level in the Pik3ca transcriptional initiation region promoted the transcription of Pik3ca, and Pik3ca promoted doxorubicin induced apoptosis in H9C2 cells by activating the PI3K/AKT pathway, providing a new theoretical basis for the study of doxorubicin cardiomyopathy.

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