利用免疫组织化学及蛋白质组学对树莓提取物抑制大鼠肝癌机制的研究

目的：探讨树莓预防大鼠原发性肝癌增殖抑制和凋亡诱导作用，寻找树莓预防大鼠原发性肝癌的特异性蛋白质靶点。方法：利用二乙基亚硝胺（DEN）建立大鼠原发性肝癌动物模型；通过免疫组织化学方法研究树莓提取物对于大鼠原发性肝癌的预防效果和形态学变化，利用蛋白质组学研究树莓预防大鼠原发性肝癌的特异性蛋白质靶点。结果：树莓提取物能抑制PCNAJVEGF的表达，抑制细胞增殖，并诱导细胞凋亡。蛋白质组学差异分析表明：成瘤组大鼠血清在蛋白质峰2597．93M／Z，4513．88M／Z上与高剂量树莓干预组大鼠血清具有显著性差异（P〈0．05）。结论：树莓提取物可以抑制肝癌细胞PCNA的表达，从而抑制肝癌细胞的增殖；还可以诱导肝癌细胞凋亡；蛋白质峰2597．93M／Z及4513．88M／Z所表达蛋白为其特异性作用靶点。

Immunohistochemistry and Proteomic Study on Chemoprevention Effect of Raspberry in Primary Liver Cancer of Rats

Objective： To investigate the preventive and inhibitive effect of phyto- chemicals within raspberry extracts on the ex- perimental primary liver cancer of rats, and to find chemoprevention mechanism of raspberry in primary liver cancer of rats. Methods： An experimental animal model of primary liver cancer in rats was established, by DEN （diethylnitr-osamine）. To explore the preventive and inhibitive effect of different doses raspberry extracts on the experi- mental primary liver cancer in rats by using the methods of immuno- histochemistry. Results： With the concentration increasing of raspberry extracts, the number of apoptotic cancer cells increased obviously. The number of apoptotic cancer cells in high-dose preventive group was significantly more than that in the control group （P 〈0.05 ）. There were significant differences at the serum protein peaks of 2597.93 M/Z and 4513.88 M/Z between rats in high-dose preventive group and those of the control group （P 〈0.05 ）, and the two protein peaks maybe the targets for chemoprevention of raspberry. Conclu- sions： Raspberry extracts can inhibit the expression of PCNA of liver cancer cells and induce the apoptosis of liver cancer cells. It is con- cluded that the phytochemicals in raspberry extracts may play a decisive role in prevention and inhibition of the liver cancer development and progression. The two serum proteins with peaks of 2597.93 M/Z and 4513.88 M/Z may be the targets for chemoprevention of raspberry.