刺参酸性粘多糖联合氟尿嘧啶对H22肝癌荷瘤小鼠的抗肿瘤作用

[Antitumor effect of Stichopus japonicus acidic mucopolysaccharide combined with fluorouracil in mice bearing neoplasia of H22 hepatoma cells].

作者信息

Chen Dandan, Song Yang, Dai Haihua

出版信息

Wei Sheng Yan Jiu. 2014 Jul;43(4):598-602.

PMID:25199288

Abstract

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.

摘要

目的

观察刺参酸性粘多糖联合氟尿嘧啶对H22肝癌荷瘤小鼠的抗肿瘤作用，并探讨其作用机制。

方法

将50只昆明小鼠右前肢腋窝皮下接种H22肝癌细胞，然后随机分为5组，包括阴性对照组、5-氟尿嘧啶组、刺参酸性粘多糖组和两个联合组。采用苏木精-伊红（HE）染色观察肿瘤组织病理变化。采用免疫组织化学和实时荧光定量PCR方法检测H22肿瘤组织中增殖细胞核抗原（PCNA）蛋白水平、P53、P21、细胞周期蛋白D1（Cyclin D1）和细胞周期蛋白依赖性激酶4（CDK4）蛋白及mRNA水平。

结果

与阴性对照组相比，其他各组H22肿瘤组织重量均有统计学意义的减轻（P<0.05）。5-氟尿嘧啶组和两个联合组的抗肿瘤率均超过50%。高剂量5-氟尿嘧啶联合刺参酸性粘多糖组的抗肿瘤率高达62.73%。HE染色显示，除阴性对照组外，其他各组肿瘤细胞减少，排列疏松，核固缩，坏死面积增加。免疫组织化学和实时荧光定量PCR结果显示，给药组PCNA蛋白、P53、Cyclin D1和CDK4蛋白及mRNA表达均较阴性对照组明显降低（P<0.05），P21蛋白表达高于阴性对照组（P<0.05），高剂量5-氟尿嘧啶联合刺参酸性粘多糖组相关基因mRNA和蛋白表达变化更显著（P<0.05）。