血管平滑肌细胞表型改变与肿瘤抑制基因p21^(WAF1)和p53的关系  

Association between the changes of vascular smooth muscle cell phenotype and tumor suppressor gene of p21^(WAF1) and p53

作　　者：闫承慧[1] 吴焱[2] 金焰[2] 王柏秋[2] 王哲[2] 张贵寅[2] 李璞[2] 傅松滨[2] 

机构地区：[1]解放军沈阳军区总医院心内科,全军心血管病研究所,辽宁省沈阳市110016 [2]哈尔滨医科大学医学遗传学研究室,黑龙江省哈尔滨市150086

出　　处：《中国临床康复》2005年第15期56-57,共2页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:增殖表型血管平滑肌细胞中肿瘤抑制基因p21WAF1基因和p53基因表达可能具有对抗血管平滑肌细胞的增殖能力,观察血管平滑肌细胞表型变化与肿瘤抑制基因p21WAF1和p53的关系。方法:实验于2003-09/10哈尔滨医科大学医学遗传学研究室完成。应用胶原酶消化法建立大鼠原代培养血管平滑肌细胞,去血清诱导体外培养血管平滑肌细胞分化。流式细胞分析检测不同表型血管平滑肌细胞增殖能力,反转录-聚合酶链反应和蛋白质印迹分析方法检测血管平滑肌细胞分化相关基因平滑肌特异性α-肌动蛋白和calponin表达变化,反转录-聚合酶链反应和蛋白质印迹杂交方法检测p21WAF1和p53基因表达变化。结果:去血清培养后,原代培养大鼠血管平滑肌细胞增殖能力下降,DNA合成前期细胞明显增加(48.38±2.35)%,细胞分化基因平滑肌特异性α-肌动蛋白和calponin明显表达。肿瘤抑制基因p21WAF1和p53表达下降。在200g/L血清培养后,大鼠血管平滑肌细胞增殖旺盛,DNA合成前期细胞相对较少(28.80±1.58)%,细胞分化相关基因平滑肌特异性肌动蛋白和calponin表达明显下调,肿瘤抑制基因p21WAF1和p53表达显著增加。结论:肿瘤抑制基因表达与血管平滑肌细胞表型状态密切相关。AIM: The proliferation of the expression of tumor suppressor genes of p21 WAF1 and p53 in phenotype vascular smooth muscle cells may have the ability against the proliferation of vascular smooth muscle cells.This paper aims to observe the association between the phenotype changes of vascular smooth muscle cells and the tumor suppressor genes of p21 WAF1 and p53. METHODS:The experiment was carried out in the Research Room of Medical Genetics, Harbin Medical University between September and October 2003.The model of rat vascular smooth muscle cells is constructed by primary culture with the collagenase digestive method, and the cells differentiation was induced in vitro by serum-deprivation.The proliferating ability of vascular smooth muscle cells of different phenotypes was detected with flow cytometry, the vascular smooth muscle cells differentiated related gene smooth muscle specific alpha-actin and calponin expression changes were detected with reverse transcription polymerase chain reaction and Western blot analysis, with which the expression of tumor suppressor genes p21 WAF1 and p53 were also detected. RESULTS: After culture of serum deprivation, the proliferation of primarily cultured vascular smooth cells was decreased, DNA presynthetic phase cells were obviously increased [(48.38%±2.35)%], and cells differentiated related gene smooth muscle specific alpha-actin and calponin expressed obviously, and the expressions of tumor suppressor genes p21WAF1 and p53 were decreased. After culture in 200 g/L serum, the proliferation of vascular smooth cells was increased, and DNA presynthetic phase cells were relatively fewer [(28.80%±1.58)%], and the expressions of cells differentiated related gene smooth muscle specific alpha-actin and calponin were obviously down-regulated, the expressions of tumor suppressor genes p21WAF1 and p53 were significantly increased. CONCLUSION: The expression of tumor suppressor genes are closely associated with the phenotype status of vascular smooth muscle cells, which indicates

关 键 词：肿瘤抑制基因 P^21 反转录-聚合酶链反应 表型改变 CALPONIN 血管平滑肌细胞增殖 血管平滑肌细胞表型 p^53基因表达 细胞增殖能力 Α-肌动蛋白 分化相关基因 哈尔滨医科大学 蛋白质印迹分析 DNA合成 WAF1基因 细胞表型变化 

分 类 号：R730.21[医药卫生—肿瘤] R735.370.2[医药卫生—临床医学]