角果木提取物tagalsin对H22细胞原位移植肝癌p53和Bcl-2表达的影响  

Effect of tagalsin on p53 and Bcl-2 expression in hepatoma H22 tumor-bearing mice

作　　者：宋修岐[1] 郭云良[2] 王炳高[1] 孙少杰[1] 姚如永[2] 

机构地区：[1]青岛大学第二附属医院普外科,266042 [2]青岛大学第一附属医院脑血管病研究所

出　　处：《中华肿瘤杂志》2011年第7期499-503,共5页Chinese Journal of Oncology

基　　金：基金项目：国家“863”项目（2006AA092446）;青岛市科技局项目（07-2-1-8-NSH-1）

摘　　要：目的探讨角果木提取物tagalsin对小鼠原位移植肝癌的干预作用。方法建立小鼠原位移植性肝癌模型，建模第10天将小鼠随机分为低、中、高tagalsin组和卡莫氟阳性对照组、食用油对照组，分别灌胃给药，给药2个疗程。期间观察各组荷瘤小鼠的生存时间、有无腹水、肝内外转移情况以及体重变化。末次给药24h后处死小鼠，计算各组小鼠的肿瘤体积、抑瘤率和脾脏指数，观察肿瘤病理组织学变化。利用免疫组化和逆转录聚合酶链反应（RT—PCR）分别从蛋白和mRNA水平检测各组p53和Bcl-2的表达水平。结果tagalsin可有效抑制原位移植肝癌生长，低、中、高剂量tagalsin组的抑瘤率分别为17．9％、63．1％和71．8％，低、中剂量tagalsin对荷瘤小鼠的脾脏指数无明显影响。病理组织学检查，各tagalsin组以及卡莫氟阳性对照组均可见细胞凋亡形态变化。免疫组化检测结果显示，卡莫氟阳性对照组及各tagalsin组突变型p53、Bcl-2细胞阳性表达率明显低于食用油对照组（P〈0．05）。RT-PCR检测结果显示，与食用油对照组相比，卡莫氟阳性对照组及各tagalsin组野生型p53基因的表达水平均升高，而Bcl-2mRNA的表达表达水平下降（P〈0．05），而以高剂量tagalsin组最为显著。结论tagalsin可能通过上调野生型p53而降低Bcl-2的表达，抑制小鼠H22细胞原位移植肝癌。Objective To explore the effect and mechanism of tagalsin on hepatoma cells. Methods The animal models were established by transplanting H22 mouse hepatoma cells to mouse liver, and ten days later the mice were randomly divided into five groups ： blank group, carmofur positive group and tagalsin groups, including low-dose, middle-dose and high-dose groups. Then medicine or oil was given to the mice by gastric gavage in consecutive 5 days with a 2-days interval as a course of treatment, two courses in all. All mice were killed at 24 hours after medication, and the survival period, ascites conditions, aggressive conditions intra- or extra- liver, weight changes, tumor volume and spleen index of the tumor- bearing mice were observed. Pathological changes of the tumors were examined. Apoptotic factors p53 and Bcl-2 protien and mRNA were detected by immunohistochemistry and reverse transcription polymerase chain reaction （RT-PCR）. Results tagalsin inhibited the hepatoma growth effectively without influencing spleen index to some extent. The tumor inhibition rate of tagalsin low, middle and high dose groups were 17.9% , 63.1% and 71.8%, respectively. Immunohistochemical results showed that the p53 and Bcl-2 protein positive cell counts of the positive control and experimental groups were significantly lower than those of the blank group （P 〈 0.01 ）. RT-PCR results showed that the p53 mRNA expression was significantly enhanced and Bel-2 mRNA expression was decreased in the positive control groups and tagalsin treatment groups, especially in the high dose group, compared with those of the blank group （P 〈 O. 05）. Conclusions tagalsin can inhibit the growth of mouse hepatoma cells significantly. The mechanism of its anti-tumor effect may work via up-regulating the wild type p53 gene expression and down-regulating Bcl-2 gene expression and thus regulating tumor cell apoptosis.

关 键 词：Tagalsin 肝肿瘤 H22细胞 免疫组化 逆转录聚合酶链反应 p53 BCL-2 凋亡 

分 类 号：R285[医药卫生—中药学]