摘要
目的探讨PTEN/PI3K/Akt信号传导通路对人慢性粒细胞白血病细胞系K562的增殖、凋亡调控的研究及可能的分子作用机制。方法将携带有野生型PTEN及绿色荧光蛋白的腺病毒(Ad-PTEN-GFP)及空载体(Ad-GFP)腺病毒,转染人慢性粒细胞白血病细胞系K562。通过MTT检测细胞生长曲线,流式细胞术检测细胞凋亡率和细胞增殖指数,同时用细胞光镜、电镜形态等方法检测细胞凋亡,荧光定量PCR(FQ-PCR)检测PTEN及凋亡相关基因Bcl-2、Bcl-xL、BaxmRNA水平变化,Western blot检测PTEN及Akt、p-Akt蛋白水平变化。结果与Ad-GFP组相比,Ad-PTEN-GFP转染K562细胞后,细胞增殖受抑,增殖指数降低,凋亡率增加,p-Akt表达降低,抗凋亡相关基因Bcl-2、Bcl-xLmRNA表达降低,促凋亡基因BaxmRNA表达增加。结论过表达PTEN可能通过抑制PI3K/Akt通路抑制K562细胞系增殖,促进细胞凋亡。
Objective To investigate the effect of PTEN/PI3K/Akt signal pathway on cell proliferation,apoptosis and its possible apoptosis-related molecular mechanism on human chronic myeloid leukemia（CML） cell line K562 cells.Methods The recombinant adenovirus containing green fluorescent protein（GFP） and PTEN（Ad-PTEN-GFP）or empty vector（Ad-GFP）was transfected into K562 cells.The growth of K562 cells was observed by MTT assay;the apoptosis rate and proliferation index（PI） were assessed by flow cytometry（FCM）.We also detected apoptosis by morphology,DNA and transmission electron microscope technique;PTEN together with anti-apoptosis gene Bcl-2,Bcl-xL mRNA and apoptosis gene Bax mRNA levels were detected by real-time fluorescent relative-quantification reverse transcriptional PCR（FQ-PCR）.PTEN and p-Akt protein levels were detected by western blotting.Results Compared with Ad-GFP growth,the growth of K562 cells transfected by Ad-PTEN-GFP was suppressed markedly,proliferation index（PI） was down regulated,apoptosis rate rised and p-Akt expression levels decreased but not total Akt;Bcl-2,Bcl-xL mRNA levels descend and Bax mRNA level increased after transfected with wild type PTEN.Conclusion Over expression PTEN gene might inhibit K562 cells proliferation and promote cell apoptosis via down regulation PI3K/Akt pathway.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2009年第10期828-832,共5页
Cancer Research on Prevention and Treatment
基金
河北省自然科学基金资助项目(C200500745)
