摘要
目的观察刺参酸性黏多糖(SJAMP)联合氟尿嘧啶(5-FU)对H22荷瘤小鼠肿瘤细胞的抑制作用,并对其作用机制进行探讨。方法 50只SPF级昆明种小鼠于右前肢腋部皮下接种H22肝癌细胞悬液,24 h后随机分为5组,分别为空白对照组(生理盐水)、5-FU组(5-FU 20 mg/kg)、SJAMP组(SJAMP 25 mg/kg)、低剂量5-FU+SJAMP组(5-FU 10 mg/kg+SJAMP 25 mg/kg)、高剂量5-FU+SJAMP组(5-FU 20 mg/kg+SJAMP 25 mg/kg)。HE染色观察各组肿瘤组织病理学改变;免疫组织化学方法检测肿瘤组织中PCNA、P53、P21、Cyclin D1及CDK4蛋白的表达水平;Real-time PCR方法检测肿瘤组织中P53、P21、Cyclin D1及CDK4 mRNA的表达水平。结果与空白对照组相比,其余4组荷瘤小鼠肿瘤实体的平均质量均明显减小(P<0.05)。5-FU组及联合用药组抑瘤率均超过50%,高剂量5-FU+SJAMP组抑瘤率高达62.73%。HE染色显示给药组肿瘤细胞排列疏松,细胞质固缩,核质比减小。免疫组化及Real-time PCR结果显示,给药组中与细胞增殖相关基因的PCNA蛋白,P53、Cyclin D1及CDK4蛋白及mRNA的表达较空白对照组中的表达明显减弱(P<0.05),P21蛋白及mRNA的表达明显增强(P<0.05),高剂量5-FU+SJAMP组相关基因mRNA及蛋白变化更加显著(P<0.05)。结论 SJAMP与5-FU均能通过下调PCNA蛋白、P53、Cyclin D1及CDK4蛋白及mRNA的表达、上调P21蛋白及mRNA的表达,达到抑制肿瘤细胞增殖的目的。SJAMP与5-FU合用具有一定的协同作用,能够明显增强抑瘤效果。
Objective To observe the antitumor effect of Stihopus japonicus acidic mucopolysaccharide combined with fluorouracil in mice bearing neoplasia of H22 hepatoma cells,and study the mechanism. Methods Inoculate the H22 liver cancer cells in the right forelimbs axillary region subcutaneous of 50 Kunming mice, and then randomly divide the mice into 5 groups, including negative control group, 5-FU group, SJAMP group and two combined groups. Tumor histopathological changes were examined by HE. The expressions of PCNA protein level, P53, P21, Cyclin D1 and CDK4 protein and mRNA levels in the H22 tumor tissues were detected by immunohistochemical and Real- time PCR methods. Results Compared with negative control group, the weights of the H22 tumor tissues in other groups are decreased statistically significant （ P 〈 0.05 ）. The anti-tumor rates of 5-FU group and the two combined groups are all exceed 50%. The anti-tumor rate of the high dose of 5-FU combined SJAMP group is reach up to 62.73%. HE staining showed that except negative control group, other groups tumor cells were less, arranged loosely , karyopyknosis and necrotic area were increased. According to the results of Immunohistochemistry and Real-time PCR, the expressions of PCNA protein, P53, Cyclin D1 and CDK4 proteins and mRNAs of the medication administration groups are decreased obviously compared with negative control group （ P 〈 0.05 ） , expressions of P21 proteins are higher than negative control group（P 〈 0.05 ） , the expressions of related genes mRNA and proteins changes are more remarkable in high dose of 5-FU combined SJAMP group （P 〈 0.05 ）. Conclusion SJAMP and 5-FU can via decrease the expressions of PCNA protein, P53, Cyclin D1 and CDK4 proteins and mRNAs, increase the expressions of P21 proteins and mRNAs, to inhibit the growth of H22 tumor in mice. And the combination of SJAMP and 5-FU shows synergistic effect which can strengthen the ability of antitumor obviously.
出处
《卫生研究》
CAS
CSCD
北大核心
2014年第4期598-602,共5页
Journal of Hygiene Research
基金
国家自然科学基金(No.81072295)
关键词
刺参酸性黏多糖
氟尿嘧啶
H22荷瘤小鼠
协同作用
Stichopus japonicus acidic mucopolysaccharide, fluorouracil, murinehepatoma H22 cell, synergistic effect
