魟鱼软骨多糖对小鼠恶性黑色素移植瘤血管生成拟态及PI3K表达的影响

Effect of ray cartilage glycosaminoglycans on vasculogenic mimicry of mice melanoma and the expression level of PI3K in melanoma of mice

目的:探讨魟鱼软骨多糖(Ray cartilage glycosaminoglycans,RCG)对小鼠恶性黑色素瘤模型中肿瘤和血管生成拟态的抑制作用及对肿瘤组织中磷脂酰肌醇3-激酶蛋白(PI3K)表达水平的影响。方法:将体外培养的B16小鼠黑色素瘤细胞接种于C57BL/6小鼠右腋下部复制肿瘤模型,将40只SPF级小鼠随机分为空白对照组(生理盐水)、环磷酰胺(CTX,60 mg/kg)组与魟鱼软骨多糖(600、300、150 mg/kg)组,观察肿瘤生长情况,计算抑瘤率;采用CD31/PAS双重染色检测魟鱼软骨多糖对肿瘤血管生成拟态计数的影响,Western Blot法测定PI3K表达变化,RT-PCR法测定PI3K基因的表达变化。结果:与空白对照组相比较,魟鱼软骨多糖的不同剂量组都对小鼠黑色素瘤的瘤重具有一定的抑制作用,且600mg/kg剂量的魟鱼软骨多糖最明显,CD31/PAS双重染色结果显示600mg/kg魟鱼软骨多糖组对血管生成生成拟态有较强的的抑制作用,Western Blot和RT-PCR结果表明随着魟鱼软骨多糖药物浓度的升高,PI3K蛋白表达水平逐渐明显降低,基因表达水平逐步明显降低,其中600mg/kg魟鱼软骨多糖组抑制作用最强。结论:魟鱼软骨多糖具有影响小鼠黑色素移植瘤和血管生成拟态生成的作用,并且可能与降低PI3K蛋白的表达有关。

Objective： To explore the Effect of RCG on the VM and weight of the Melanoma and the effect of it on the expression level of phosphoinositide 3-kinase（ PI3K）. Methods： B16 melanoma cells were cultured. The C57BL/6 mouse melanoma transplant tumor model was made with B16 melanoma cell suspending liquid inoculated in right oxter of C57BL/6 mice. The 40 experimental mice were randomly divided into control group（ isovolumic saline）,CTX group（ 60 mg/kg） and RCG high-dose（ 600 mg/kg）,medium-dose（ 300mg/kg） and low-dose groups（ 150 mg/kg）,To observe the effect of ginsenoside RCG on tumor growth and measure the inhibitory rate of tumor; CD31/PAS doublestaining show the morphology of VM in the different RCG groups,the expression of PI3 K was detected by Western Blot,the expression of PI3 K mRNA was detected by RT-PCR. Results： Compared with control group,tumor weight were lost,inhibitory rate of primary tumor in 600mg/kg RCG group was highest（ P〈0. 01）; the VMD of the VM in 600mg/kg RCG group was lowest,Western Blot and RT-PCR results show that with increasing concentration of RCG,the expression level of PI3 K protein decreased gradually（ P〈0. 01）,the expression level of PI3 K mRNA decreased gradually（ P〈0. 01）,and in both of them,the 600mg/kg RCG group had the strongest Inhibition. Conclusion： RCG could inhibit the growth of melanoma of mice,which is probably associated with that RCG reduce the protein expression of PI3 K in the tumor cells.