石见穿多糖通过PTEN通路对骨肉瘤小鼠的抑瘤作用及对免疫功能的影响

The anti-tumor effect of polysaccharides from salvia chinensis on osteosarcoma mice and its effect on immune function through PTEN pathway

关键词:  石见穿多糖  第10染色体同源丢失性磷酸酶-张力蛋白基因  骨肉瘤  免疫功能  移植瘤体积  肿瘤细胞凋亡率

Key words:  polysaccharides from salvia chinensis  phosphatase and tensin homolog deleted on chromosome ten  osteosarcoma  immune function  transplanted tumor volume  tumor cell apoptosis rate

摘要:

[目的] 探讨石见穿多糖（PSSC）通过第10染色体同源丢失性磷酸酶-张力蛋白基因（PTEN）通路对骨肉瘤小鼠的抑瘤作用及对免疫功能的影响。[方法] 选取60只BALB/C雌性裸鼠，取12只作为对照组，剩余48只采用MG-63人骨肉瘤细胞注射法建立骨肉瘤小鼠模型，成功建模42只，随机分为模型组（11只，腹腔注射生理盐水10 mL/kg）、PSSC低剂量组（11只，腹腔注射PSSC 10 mg/kg）、PSSC高剂量组（10只，腹腔注射PSSC 100 mg/kg）、顺铂组（10只，腹腔注射顺铂2 mg/kg），干预2周，绘制小鼠肿瘤生长曲线观察移植瘤体积变化，流式细胞仪测定小鼠外周血CD4⁺、CD3⁺T细胞比例，测定小鼠移植瘤质量，苏木精-伊红（HE）染色观察肿瘤病理组织变化，统计肿瘤细胞凋亡率，蛋白免疫印迹（Western Blot）法检测移植瘤组织PTEN、磷酸肌醇3-激酶（PI3K）、磷酸化蛋白激酶B（p-Akt）蛋白表达。[结果] PSSC能降低骨肉瘤小鼠移植瘤体积及质量，提高外周血CD4⁺、CD3⁺T细胞比例，减轻肿瘤病理组织变化，提高肿瘤细胞凋亡率，促进PTEN蛋白表达，抑制PI3K、p-Akt蛋白表达，且具有剂量依赖性，高剂量PSSC抑瘤作用更为明显，肿瘤抑制效用与顺铂化疗效用相当。[结论] 采用PSSC治疗骨肉瘤小鼠，能提高小鼠免疫能力，减轻肿瘤病理变化，促进肿瘤细胞凋亡，抑制移植瘤体积及质量，可能与PSSC可激活抑癌基因PTEN有关。

Abstract:

[Objective] To explore the anti-tumor effect of polysaccharides from salvia chinensis (PSSC) through phosphatase and tensin homolog deleted on chromosome ten (PTEN) pathway on osteosarcoma mice and its influence on immune function.[Methods] The 60 BALB/C female nude mice were selected,12 were selected as the control group,and the remaining 48 were injected with MG-63 human osteosarcoma cells to establish an osteosarcoma mouse model,and 42 mice were successfully modeled. The modeled mices were randomly divided into model group (11 rats,intraperitoneal injection of physiological saline 10 mL/kg),PSSC low-dose group (11 rats,intraperitoneal injection of PSSC 10 mg/kg),and PSSC high-dose group (10 rats,intraperitoneal injection of PSSC 100 mg/kg),cisplatin group (10 rats,intraperitoneal injection of cisplatin 2 mg/kg),intervention for 2 weeks. The mouse tumor growth curve was drew to observe the changes of transplanted tumor volume,the proportion of CD4⁺ and CD3⁺ T cells in peripheral blood of mice was measuredby flow cytometry,the mass of mouse transplanted tumors was determined,the changes of tumor pathological tissues was observed by HE staining,the apoptosis rate of tumor cells was counted,Western blot method was used to detect the expression of PTEN,PI3K and p-Akt protein in transplanted tumor tissue.[Results] PSSC can reduce the volume and quality of transplanted tumors in osteosarcoma mice,increase the ratio of CD4⁺ and CD3⁺ T cells in peripheral blood,reduce tumor pathological changes,increase tumor cell apoptosis rate,promote PTEN protein expression,and inhibit PI3K and p-Akt protein expression,and it is dose-dependent,high-dose PSSC has a more obvious anti-tumor effects,and tumor suppression effect is equivalent to cisplatinization effect.[Conclusion] The use of PSSC to treat osteosarcoma mice can improve the immune capacity of mice,reduce tumor pathological changes,promote tumor cell apoptosis,and inhibit the volume and quality of transplanted tumors,which may be related to the activation of the tumor suppressor gene PTEN pathway by PSSC.