鹽誘導激酶2對電離輻射誘導的自噬與凋亡的影響
發(fā)布時間:2018-03-06 07:01
本文選題:鹽誘導激酶(SIK) 切入點:電離輻射 出處:《輻射防護》2017年03期 論文類型:期刊論文
【摘要】:為探索鹽誘導激酶2(salt-inducible kinase 2,SIK2)對電離輻射引起的自噬與凋亡的影響,以期為腫瘤的放射治療提供新的思路,以慢病毒顆粒LV-GFP和LV-SIK2體外轉(zhuǎn)染He La細胞,48 h后用熒光顯微鏡觀察轉(zhuǎn)染是否成功,RT-PCR和Western blot檢測轉(zhuǎn)染細胞SIK2的干擾效果。通過Western blot檢測SIK2低表達對8Gy60Coγ射線照射下細胞中自噬相關(guān)蛋白LC3-Ⅱ和P62表達的影響來觀察自噬的變化;利用流式細胞術(shù)來檢測SIK2低表達對8 Gy60Coγ射線照射下細胞凋亡的影響。結(jié)果表明,SIK2低表達的穩(wěn)轉(zhuǎn)細胞系被成功構(gòu)建;8Gyγ射線照射后,SIK2低表達細胞系LC3-Ⅱ表達下調(diào),P62表達上調(diào),表明自噬被減弱;流式細胞術(shù)檢測發(fā)現(xiàn)SIK2低表達細胞系在照后12 h FITC+PI-細胞比例就顯著增加,表明SIK2低表達增加了細胞凋亡。因此,SIK2低表達會導致電離輻射引起的細胞自噬大大減弱,凋亡顯著增加。
[Abstract]:In order to explore the effect of salt-inducible kinase 2SIK2) on autophagy and apoptosis induced by ionizing radiation, we hope to provide a new way of thinking for radiotherapy of tumor. He-La cells were transfected with lentivirus particles LV-GFP and LV-SIK2 for 48 h, then the interference effect of SIK2 was detected by RT-PCR and Western blot with fluorescence microscope. The low expression of SIK2 was detected by Western blot. The cells were irradiated with 8 Gy 60Co 緯 -rays by Western blot. The changes of autophagy were observed by the expression of LC3- 鈪,
本文編號:1573783
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