松乳菇多糖通过PI3K/Akt信号通路对人骨肉瘤细胞MG-63裸鼠移植瘤生长的抑制作用

Inhibitory Effect of Polysaccharide from Lactarius Deliciosus( L. ex Fr.) Gray on the Growth of Xenograft Tumor of Human Osteosarcoma MG-63 in Nude Mice through PI3K/Akt Signaling Pathway

目的:研究松乳菇多糖对裸鼠皮下移植瘤生长、凋亡及相关蛋白表达的影响。方法:采用MG-63细胞建立裸鼠皮下移植瘤,随机分为5组:模型对照组、阳性对照组(氟尿嘧啶,20 mg·kg-1)、松乳菇多糖低(10 mg·kg-1)、中(20 mg·kg-1)、高(30 mg·kg-1)剂量组,每组10只。各组腹腔注射给药,模型对照组给予等量0.9%Na Cl溶液,连续14 d。每日观察各组裸鼠生活状态和肿瘤生长情况,给药第14天后,处死裸鼠,剥瘤称质量并计算抑瘤率;TUNEL法检测肿瘤组织细胞凋亡;Western blot法检测瘤组织中磷脂酰肌醇3-激酶(PI3K)、磷酸化蛋白激酶B(p-Akt)和蛋白激酶B(Akt)含量。结果:与模型对照组相比,松乳菇多糖各组裸鼠的饮食状况、活动度和精神状态均较好;瘤重明显降低(P<0.05或P<0.01);凋亡率明显增加(P<0.01);PI3K、Akt和p-Akt蛋白表达均明显下调(P<0.05或P<0.01)。与阳性对照组相比,松乳菇多糖各组裸鼠的饮食状况、活动度和精神状态无明显差异;松乳菇多糖低剂量组的抑瘤率和凋亡率均相对较低(P<0.05或P<0.01);高剂量组的凋亡率相对较高(P<0.05);PI3K、Akt和p-Akt蛋白表达差异无统计学意义(P>0.05)。结论:松乳菇多糖通过PI3K/Akt信号通路促进骨肉瘤移植瘤裸鼠肿瘤细胞凋亡。

Objective:To study the effect of polysaccharide from Lactarius deliciosus(L.ex Fr.)Gray on the growth,apoptosis and related protein expression of subcutaneous transplanted tumors in nude mice.Methods:The subcutaneous transplanted tumors in nude mice were established using MG-63 cells,and randomly divided into 5 groups:the model control group,the positive control group(fluorouracil,20 mg·kg-1),polysaccharide from Lactarius deliciosus(L.ex Fr.)Gray low(10 mg·kg-1),medium(20 mg·kg-1)and high(30 mg·kg-1)dosage groups with 10 rats in each group.Each group was given intraperitoneal injection,and the model control group was given the same amount of 0.9%Na Cl solution for 14 days.The living status and tumor growth of nude mice were observed daily.After the 14-day administration,the nude mice were sacrificed,the tumor weight was stripped,and the tumor inhibition rate was calculated.TUNEL was utilized for measuring the apoptosis.Western blot was utilized for measuring the contents of phosphatidylinositol3-kinase(PI3 K),phosphorylated protein kinase B(p-Akt)and protein kinase B(Akt)in tumor tissues.Results:Compared with that in the model control group,the diet,activity and mental state of nude mice in polysaccharide from Lactarius deliciosus(L.ex Fr.)Gray groups was better,and the tumor weight was reduced(P<0.05 or P<0.01),the apoptosis rate was increased(P<0.01),and the expressions of PI3 K,Akt and p-Akt were down-regulated(P<0.05 or P<0.01).Compared with the positive control group,there were no significant differences in diet,activity and mental state between the groups,the tumor inhibition rate and apoptosis rate of polysaccharide from Lactarius deliciosus(L.ex Fr.)Gray low dosage group were relatively lower(P<0.05 or P<0.01),the tumor inhibition rate and apoptosis rate of high dosage group was relatively higher(P<0.05),and there were no significant differences in the expressions of PI3 K,Akt and p-Akt proteins(P>0.05).Conclusion:Polysaccharide from Lactarius deliciosus(L.ex Fr.)Gray promots the apoptosis of tumor cells in nude mice transplanted with osteosarcoma through PI3 K/Akt signaling pathway.