Exon 8-defient SIRT1 splicing variant opposes SIRT1 full-length in regulating oxidative damage of 293T cells#br#

YANG Xiao-Rong; GAO Rui-Jun; LI Yan-Li; YIN Li-Tian; ZHANG Ce

doi:10.16098/j.issn.0529-1356.2020.06.008

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Acta Anatomica Sinica ›› 2020, Vol. 51 ›› Issue (6) : 855-860. DOI: 10.16098/j.issn.0529-1356.2020.06.008

Cell and Molecules Biology

Exon 8-defient SIRT1 splicing variant opposes SIRT1 full-length in regulating oxidative damage of 293T cells#br#

YANG Xiao-rong* GAO Rui-jun LI Yan-li YIN Li-tian ZHANG Ce

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Abstract

Objective To investigate the different effects of exon 8-de cient silent information regulator 1(SIRT1) splicing variant (SIRT1-ΔExon8) and SIRT1 full-length (SIRT1-FL) on cellular oxidative damage. Methods A human embryonic kidney 293T cell line was infected with SIRT1-ΔExon8 shRNA expression vectors and SIRT1-FL overexpression vectors, respectively. Administration of hydrogen peroxide(H2O2) into 293T cells was performed to induce oxidative stress injury including the changes of cell viability, lactate dehydrogenase (LDH) activity, apoptotic rate and reactive oxygen species (ROS) level as measured by cell counting kit 8(CCK-8) assay, LDH test, and flow cytometry. The mRNA levels of SIRT1-FL and SIRT1-ΔExon8 were determined by Real-time PCR. Results In a dose-dependent manner, 50-800 μmol/L H2O2 induced oxidative damage including a decrease in cell viability, an increase in LDH release, cell apoptosis and ROS level; and also caused a decrease in SIRT1-FL mRNA level and an increase in SIRT1-ΔExon8 mRNA level. In addition, overexpression of SIRT1-FL or interference of SIRT1-ΔExon8 partly inhibited H2O2 (400 μmol/L)-induced stress injury. Conclusion SIRT1-ΔExon8 may promote cellular oxidative stress injury, while SIRT1-FL plays an opposite role.

Key words

Hydrogen peroxide / Oxidative stress / Silent informaton regulator 1-full-length / Silent informaton regulator 1-ΔExon8 / Flow cytometry

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YANG Xiao-rong GAO Rui-jun LI Yan-li YIN Li-tian ZHANG Ce. Exon 8-defient SIRT1 splicing variant opposes SIRT1 full-length in regulating oxidative damage of 293T cells#br#[J]. Acta Anatomica Sinica. 2020, 51(6): 855-860 https://doi.org/10.16098/j.issn.0529-1356.2020.06.008

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