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目的研究昆参颗粒对S180荷瘤小鼠C-myc、P16蛋白表达及P16基因甲基化的影响。方法将昆明种小鼠60只以数字表法随机分为模型对照组、参莲胶囊组、昆参大、小剂量组,每组15只。全部小鼠于右侧腋窝下接种S180瘤株,每鼠0.2ml。各组在接种瘤株后第3天进行灌胃给药,每日1次。昆参大剂量组(昆参颗粒800mg/kg)、昆参小剂量组(昆参颗粒400mg/kg)、参莲胶囊组(参莲胶囊600mg/kg)、模型对照组以等量生理盐水灌胃。第12天处死所有小鼠,剥离肿瘤组织。Methylight法检测小鼠胃癌组织P16基因甲基化。免疫组织化学(S-P法)测定肿瘤组织中C-myc、P16蛋白表达。结果与模型对照组比较,昆参颗粒能明显抑制P16基因的甲基化、提升P16蛋白表达水平(P<0.05)。昆参颗粒抑制P16基因的甲基化、增加P16蛋白表达的作用优于参莲胶囊,且随剂量的增加而增大。昆参颗粒对C-myc蛋白表达的抑制作用随剂量的增加而增大。以上结果显示:昆参颗粒能明显抑制P16基因的甲基化、提升P16蛋白表达水平并抑制C-myc蛋白表达,存在一定的量效关系,起效剂量为400mg/kg。小鼠胃组织中P16与C-myc蛋白表达呈负相关(P<0.05)。结论昆参颗粒可抑制P16基因的甲基化,提高胃癌组织P16蛋白的表达水平,同时抑制C-myc蛋白表达可能是其发挥抗肿瘤功效的重要机制。
Objective To study the effect of Kun Shen Granule on the expression of C-myc, P16 protein and methylation of P16 gene in S180 tumor-bearing mice. Methods Sixty Kunming mice were randomly divided into model control group, Shenlian Capsule group and Kunming large and small dose groups, with 15 in each group. All mice were inoculated with the S180 tumor strain on the right armpit, 0.2 ml per mouse. Each group on the 3rd day after inoculation of tumor strains for intragastric administration, 1 day. Kunqushen high dose group (KunShen Granules 800mg / kg), KunShen low dose group (KunShen Granules 400mg / kg), ShenLian capsule group (ShenLian capsule 600mg / kg), the model control group with the same amount of saline irrigation stomach. All mice were sacrificed on day 12 and the tumor tissue dissected. Methylight Methylation Detection of P16 Gene in Mouse Gastric Cancer. Immunohistochemistry (S-P method) was used to detect the expression of C-myc and P16 in tumor tissue. Results Compared with the model control group, KunShen Granule could significantly inhibit the methylation of P16 gene and increase the expression of P16 protein (P <0.05). KUNSENG granules inhibited the methylation of P16 gene and increased the expression of P16 protein, which was better than that of SHENLIAN capsule, and increased with the increase of dosage. The inhibitory effect of Kun Shen Granules on C-myc protein expression increased with increasing dose. The above results show that KunShen granules can significantly inhibit the methylation of P16 gene, enhance the expression of P16 protein and inhibit the expression of C-myc protein, there is a certain dose-effect relationship, the effective dose is 400mg / kg. There was a negative correlation between P16 and C-myc protein expression in gastric mucosa (P <0.05). Conclusion KUNSENG Granule can inhibit the methylation of P16 gene, increase the expression of P16 protein and inhibit the expression of C-myc protein in gastric cancer tissue, which may be an important mechanism of its anti-tumor effect.