摘要
目的观察β-淀粉样肽(25-35)〔β-amyloidpeptide(25-35),Aβ25-35〕对体外血清饥饿培养PC12细胞的CyclinD1、CDK4、pRb、E2F1基因表达的影响。方法用终浓度为25μmol/LAβ25-35处理PC12细胞,流式细胞仪检测分析细胞周期的改变,通过RT-PCR检测CyclinD1、CDK4、E2F1基因mRNA表达变化,Western印迹检测CyclinD1、CDK4、pRb蛋白表达的变化。结果流式细胞仪分析表明血清饥饿培养24h可使约90%PC12细胞停滞于G0/G1期,25μmol/LAβ25-35诱导组8、16、24h与对照组比较,S期百分率明显增加(P<0.01),16h后细胞凋亡率明显增加(P<0.01),可见明显的亚二倍体峰(Ap峰);Aβ25-35浓度诱导血清饥饿培养的PC12细胞0～20h,CyclinD1、CDK4、pRb、E2F1mRNA和蛋白表达增高。结论Aβ25-35诱导同步化于G0/G1的PC12细胞重新进入细胞周期,并阻滞于S期,同时出现凋亡,可能与增加CyclinD1、CDK4、pRb、E2F1mRNA和蛋白的表达有关。
Objective To study the effects of β-amyloid peptide （25 - 35 ） （Aβ25-35） on the expression of gene cyclin D1, CDK4, pRb, E2F1 in starvation cultured PC12 cell by in vitro serum. Methods PC12 cells were treated with 25μmol/L Aβ25-35. Cell cycle distribution was analyzed by flow cytometry （FCM） , mRNA expressions of cyclin D1, CDK4 and E2F1 were detected by RT-PCR and protein expressions of cyclin D1, CDK4 and pRb were detected by Western-blot. Results About 90% PC12 cells were found arrest on G0/G1 phase by FCM at 24 h starvation cultured by serum. Compared with control group, after treated with 25 9 moL/L Aβ25-35 for 8, 16, 24 h, the percent of S phase cells were raised remarkably （P 〈 0. 01 ） and the rate of apoptotie cells at treated with 25 μmol/L Aβ25-35 16 h was increased obviously （P 〈0. 01 ） and apparent hypodiploid peak （ Ap peak） was observed. Treated with 25 μmol/L Aβ25-35 from 0 to 20 h,the mRNA and protein expressions of Cyclin Dl , CDK4, pRb and E2F1 gene were upregulated. Conclusions PC12 cells induced by Aβ25-35 synchronized G0/G1 phase can reenter cell cycle, be attested on S phase, and appear apoptotic peak subsequently, which maybe associated with Aβ25-35＇s increasing the mRNA and protein expressions of cyclin D1, CDK4, pRb and E2FI gene.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2009年第4期418-421,共4页
Chinese Journal of Gerontology
基金
广东省重点学科项目(0508)
广东省中医药局项目(2007129)
湛江市科技计划项目资助(2004003)
