Analysis of autophagy related protein p62 in carcinogenesis of oral cancer

一、体外分析本研究的目的是揭示自噬相关蛋白p62与细胞凋亡和口腔癌发生的关系。流式细胞仪分析表明，p62(-/-)MEF增强了对细胞凋亡的抵抗力，siRNA实验也得到了类似的结果。实时荧光定量聚合酶链式反应显示Bcl2和Bclxl表达增强。免疫印迹分析显示，经UVB照射后，野生型MEF和p62(-/-)细胞中Bcl2、Bclxl和Bax的表达均降低，而野生型MEF和p62(-/-)MEF中Bcl2、Bclxl和Bax的表达无明显差异。P62(-/-)和野生型MEF的ATM基因表达差异无统计学意义。UVB照射p62(-/-)和野生型小鼠皮肤的体内分析表明，p62(-/-)的皮肤具有较低的抗凋亡能力。我们用p62(-/-)和Nrf2(-/-)小鼠建立了NASH模型小鼠。

I. in vitro analysis The purpose of this study is to reveal the association of autophagy related protein p62 and apoptosis, and carcinogenesis of oral cancer.FACS analysis revealed that p62 (-/-)MEF increased resitance to apoptosis, and the similar result was obtained from siRNA experiment.Stat3 and Src phosphorylation were increased in p62 (-/-)MEF. Real time PCR revealed Bcl-2 and Bcl-xL expression were increased. Immuno-blotting analysis showed that Bcl-2, Bcl-xL, and Bax were reduced in p62 (-/-) and wild type MEF by UVB irradiation, however, no significant difference were observed between p62 (-/-) and wild type MEF. ATM mRNA expression were not different between p62 (-/-) and wild type MEF.II. in vivo analysis By UVB irradiation to skin of p62 (-/-) and wild typ mice showed that p62 (-/-) skin had less apoptotic response resulting resistance to apoptosis.III. Additional results Prx I (-/-) MEF showed reduced resistance to apoptosis. We found NASH model mouse by using p62 (-/-) and Nrf2 (-/-) mice.