健脾理气方对HepG2人肝癌细胞株p53和bcl-2基因表达的影响

Effect of Chinese meidicine on exprecion of p53 and bcl-2 in cultured HepG-2 cells

[目的]观察健脾理气方对HepG2人肝癌细胞株p53和bcl-2基因表达的影响。[方法]30只雄性SD大鼠随机分为健脾理气方(中药)组、0.85%氯化钠(对照)组和顺铂(化疗)组,每组10只。利用血清药理学方法制备药物血清,3组分别加入PRMI 1640血清培养的HepG2人肝癌细胞株中,作用48 h。应用免疫细胞化学方法和图像分析方法观察健脾理气方对HepG2人肝癌细胞株p53、bcl-2基因表达影响。[结果]①免疫细胞化学法显示抑癌基因p53主要定位在细胞核,癌基因bcl-2主要定位在细胞质内。药物血清作用后,p53表达明显增强;bcl-2表达明显降低。随着作用时间延长,2种基因表达改变更明显。②中药组与对照组p53和bcl-2光度(AOD)值差异有统计学意义(P<0.01)。③化疗组作用结果与中药组相似。[结论]健脾理气方药物血清上调p53、下调bcl-2,达到抑制HepG2人肝癌细胞株的作用。为应用健脾理气方治疗肝癌提供实验依据。

[Objective]To research the expression of p53, bcl-2 gene by Jianpi Liqi herbs on HepG-2 cells. [Methods]Three groups were divided： normal saline group, isotonic Na chloride group and cisplatin chemotherapy group. Medecine serum was prepared by serum pharmacological methods. HepG2 cells were cultured in PRMI1640 serum. Related medeeine serum were added into the three groups respectively and were co-cultured for 48 hours. Immunocytochemistry and image analysis methods were applied to detect the expression of p53, bcl-2 gene by Jianpi Liqi herbs on HepG-2 cells. [Results] （1）Immunoeytochemistry method showed that anti-oncogene p53 was mainly located in the nucleus and oncogene bcl-2 in the cytoplasm. After the effect of drug serum, p53 expression increased obviously and bcl-2 expression was significantly lower. With the extension of the time,the changes of two types of gene expression were more obvious. （2）Compared with the control group, the value AOD of p53 and bcl-2 in the drug serum group had significant difference, P〈0. 01. （3）The effect of Cisplatin chemotherapy group was similar to drug serum group, [Conclusion]Drug serum of Jianpi Liqi herbs can up-regulate p53, down-regulate bcl-2 and achieve the suppression of HepG2 cell.