摘要
目的:探讨百里醌对血管生成和胰腺癌生长的影响及其机制。方法:采用贴壁选择法培养人脐血内皮祖细胞(EPCs),细胞免疫组化检验细胞中血管内皮生长因子受体-2(VEGFR-2)、VIII因子和CD34表达,证实细胞属性;观察不同浓度(10 nmol/L、20 nmol/L和40 nmol/L)百里醌对EPCs小管形成的影响;不同浓度(20μmol/L、40μmol/L和80μmol/L)百里醌作用人胰腺癌细胞株PANC-1后,Western blotting检测胰腺癌细胞中血管内皮生长因子(VEGF)表达的变化;建立裸鼠胰腺癌原位移植瘤模型,分成对照组和百里醌组,实验结束后观察百里醌对裸鼠胰腺癌生长的抑制作用;免疫组织化学法检测裸鼠胰腺肿瘤组织中Ki-67、CD34和VEGF的阳性表达。结果:体外成功培养出人脐血EPCs,百里醌可显著抑制体外EPCs小管形成;并抑制体外人胰腺癌PANC-1细胞中VEGF的表达;与对照组相比较,百里醌可明显抑制荷瘤裸鼠中胰腺肿瘤生长,并下调Ki-67、CD34和VEGF在胰腺肿瘤组织中的阳性表达。结论:百里醌可抑制体内外血管生长,有望作为治疗胰腺癌的血管抑制药物。
AIM： To investigate the anti-angiogenic effect of thymoquinone on pancreatic cancer.METHODS： The endothelial progenitor cells（EPCs） isolated from human umbilical cord blood were cultured in vitro through adhesion selection and were differentiated into endothelial cells with the induction of special cytokines.The endothelial cell lineage was confirmed by the positive expression of VEGFR-2,Ⅷ factor and CD34.A tube formation assay was performed after EPCs were cultured with various concentrations of thymoquinone（10 nmol/L,20 nmol/L and 40 nmol/L）.Human pancreatic cancer cell line PANC-1 was treated with different concentrations of thymoquinone（20 μmol/L,40 μmol/L and 80 μmol/L）.The protein expression of VEGF in PANC-1 cells was detected by Western blotting.Metastatic model of human pancreatic cancer was established by orthotropic implantation of histologically intact human tumor tissue into the pancreatic wall of nude mice.After 2 weeks of implantation,the mice were randomized into 2 groups（n=10）： vehicle alone（control） and thymoquinone（20 mg/kg given daily by intragastric intubation for 14 days）.Eight weeks after implantation,tumor weight and inhibitory rate were evaluated after the mice were sacrificed.The expression of Ki-67,CD34 and VEGF in the tumors was determined by the method of immunohistochemistry.RESULTS： The EPCs isolated from human umbilical cord blood were successfully cultured in vitro.Incubation of EPCs with thymoquinone decreased the tube-forming capacity of EPCs in a concentration-dependent manner.Thymoquinone down-regulated the protein expression of VEGF in PANC-1 cells.Compared with control group,the final tumor weight showed a significant decrease in thymoquinone group.Furthermore,the percentages of Ki-67,CD34 and VEGF-positive cells in the tumors were significantly reduced by thymoquinone treatment.CONCLUSION： Thymoquinone,which inhibits angiogenesis both in vitro and in vivo,may be an anti-angiogenic drug for treating pancreatic cancer.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2011年第12期2281-2285,共5页
Chinese Journal of Pathophysiology
基金
温州市科技局资助项目(No.Y20090121)
关键词
胰腺肿瘤
百里醌
内皮祖细胞
Pancreatic neoplasms
Thymoquinone
Endothelial progenitor cells
