温下方抑制肺癌A549细胞增殖及对PCNA Rb CyclinD1表达影响.doc

温下方抑制肺癌A549细胞增殖及对PCNA Rb CyclinD1表达影响摘 要:目的:研究温下方对肺癌A549细胞增殖的抑制作用及对PCNA、Rb、CyclinD1表达的影响。方法:正常血清及不同浓度的温下方含药血清作用于A549细胞,MTT法检测含药血清对A549细胞的抑制作用,激光共聚焦显微镜观察PCNA、Rb、CyclinD1的表达。结果:温下方大、小剂量含药血清组作用24h抑制率为29.85%、25.90%,作用36h抑制率为42.02%、35.10%;PCNA表达降低至3.08±0.97、3.15±0.88,CyclinD1表达降低至2.65±0.83、2.70±0.79。结论:温下方可明显抑制肺腺癌细胞的增殖,明显降低PCNA、CyclinD1表达。 关键词:温下方;A549;PCNA;Rb;CyclinD1? 中图分类号:R285.5 文献标识码:A 文章编号:1673-7717(2010)04-0721-03 Wenxia Formular Restrain the Multiplication of the A549 Cells and the Infection to the Expression of PCNA、Rb and CyclinD1 JI Xu?ming, OUYANG Bing, WANG Chun?yan (Shandong University of TCM, Jinan 250014, Shandong, China) Abstract:Objectives: To investigate the restrain of Wenxia Formular to the multiplication of A549 cells and the influence to the expression of PCNA、Rb and CyclinD1. ?Methods: ?A549 cell lines were cultured with normal serum and different concentration of serum containing wenxia Formular. the MTT was used to exam the inhibition rate to A549 cells,he expression of PCNA、Rb and CyclinD1 in cells were evaluated by Laser Scanning Confocal Microscope.?Results:? After A549 cells 24h and 36h treated with wenxiafang big and small dose medicated serum,the rates of restrain were 29.85% and 25.90%,42.02% and 35.10%,The expression of PCNA and CyclinD1 significantly suppressed to 3.08±0.97,3?15±0.88 and 2.65±0.83,2.70±0.79(P 2.4 激光共聚焦显微镜法测PCNA Rb cyclinD1蛋白表达 采用免疫荧光细胞化学法。PCNA、Rb、cyclinD1抗体的效价均为1∶100。标本PBS洗3次,山羊血清室温孵育15min,分别加入一抗,4℃过夜;PBS洗3次,加入荧光素标记的二抗,室温孵育20min;PBS洗3次。激光共聚焦显微镜进行分析[2],表达强度用平均荧光强度表示。 2.5 数据分析采用SPSS 11.5软件包进行统计分析,数据用均数±标准差(±s)表示,计量资料用配对?t?检验,计数资料用χ2检验。 3 结 果 3.1 含药血清对肺癌A549细胞生长的抑制作用 表1含药血清对肺癌细胞生长的抑制作用 (%,±s) 注:与对照组比较, *P 1