摘要
目的探讨蛇毒半胱氨酸蛋白酶抑制剂(snake venom cystatin,sv-cystatin)重组蛋白对小鼠黑色素瘤血管生成拟态(vasculogenic mimicry,VM)形成的影响及相关机制。方法建立Matrigel胶肿瘤细胞类血管生成模型,分析sv-cystatin重组蛋白(10、25、50、100、200mg/L)对B16F10细胞VM形成及基质金属蛋白酶-2、9[matrix metalloproteinase-2(MMP-2)、MMP-9]蛋白表达的影响;建立C57BL/6小鼠黑色素瘤实验性肺转移模型,经25、50mg/kg重组蛋白治疗后,应用CD34和过碘酸雪夫(periodic acid-schiff stain,PAS)双重染色法检测肺转移瘤VM形成,免疫组织化学法检测转移瘤MMP-2、MMP-9表达。结果与对照组相比,sv-cystatin重组蛋白作用后B16F10细胞体外VM形成及MMP-2、MMP-9蛋白表达明显减少,并呈剂量依赖性(P<0.05);两治疗组小鼠肺转移瘤VM形成及MMP-2、MMP-9表达明显下降(P<0.05)。结论 sv-cystatin重组蛋白能够抑制VM形成;下调MMP-2、MMP-9表达是其中的机制之一。
Objective To investigate the effect of recombinant snake venom cystatin （sv-cystatin） on vasculogenic mimicry （VM） formation of mouse melanoma in vitro and in vivo and the mechanisms underlying this effect. Methods The effect of recombinant sv-cystatin （10, 25, 50, 100 and 200 mg/L） on VM of mouse melanoma cell B16F10 in vitro was observed by tube-like structure formation. Experimental lung metastasis assay was used to analyze the effect of sv-cystatin （three intravenous injections of 25 or 50 mg/kg） on VM in surface metastatic lesions by CD34 and periodic acid-Schiff （PAS） double staining. Matrix metalloproteinase-2 （MMP-2） and matrix metalloproteinase-9 （MMP-9） expressions in B16F10 cells were assessed by immunocytochemistry in vitro and immunohistochemistry in vivo, respectively. Results The number of VM and the expressions of MMP-2 and MMP-9 were decreased significantly in dose-dependent manner after treatment with an increased dose of recombinant sv-cystatin compared to those in control group （P （0. 05）. In vivo, recombinant sv-cystatin administration significantly suppressed lung tumor colonies, VM formation as well as MMP-2 and MMP-9 expressions in C57BL/6 mice （P〈0.05）. Conclusion Recombinant sv-cystatin can inhibit VM formation of B16F10 cells via down-regulating the expressions of MMP-2 and MMP-9.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2014年第2期169-174,共6页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金资助项目(No.30371747)
福建省科技重大资助项目(No.2002Y003)
福建省自然科学基金指导性科技计划项目(No.2012D117)~~
