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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 inhibitory effect of SJAMP combined with 5-FU on tumor cells in H22 tumor-bearing mice and to explore its mechanism. Methods Fifty Kunming SPF Kunming mice were inoculated subcutaneously with H22 hepatoma cells in the axilla of the right forelimb and were randomly divided into 5 groups after 24 h: blank control group (saline), 5-FU 20 (SJAMP 25 mg / kg), low dose 5-FU + SJAMP group (5-FU 10 mg / kg + SJAMP 25 mg / FU 20 mg / kg + SJAMP 25 mg / kg). The expression of PCNA, P53, P21, Cyclin D1 and CDK4 protein in tumor tissue were detected by immunohistochemical method. The expression of P53, P21, Cyclin D1 in tumor tissue was detected by Real-time PCR CDK4 mRNA expression levels. Results Compared with the blank control group, the average tumor mass of the other four groups of tumor-bearing mice were significantly reduced (P <0.05). The inhibition rates of 5-FU group and combination group were more than 50%, and the inhibition rate of high-dose 5-FU + SJAMP group was 62.73%. Hematoxylin-eosin staining showed that the tumor cells in the treatment group were loosely arranged, cytoplasm was contracted, and the ratio of nuclear to cytoplasm was decreased. The results of immunohistochemistry and real-time PCR showed that the expressions of PCNA, P53, Cyclin D1 and CDK4 protein and mRNA in the cell proliferation-related genes in the treated group were significantly weaker than those in the blank control group (P <0.05) P21 protein and mRNA expression was significantly increased (P <0.05), high-dose 5-FU + SJAMP group mRNA and protein changes more significant (P <0.05). Conclusion Both SJAMP and 5-FU can up-regulate the expression of P21 protein and mRNA by downregulating the expression of PCNA protein, P53, Cyclin D1 and CDK4 protein and mRNA, thereby inhibiting the proliferation of tumor cells. SJAMP combined with 5-FU has a certain synergistic effect, can significantly enhance the anti-tumor effect.