摘要
目的:探讨恶性胶质瘤组织中糖酵解限速酶6-磷酸果糖激酶-2/果糖双磷酸酶-2同工酶3(6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3,PFKFB3)的表达水平及PFKFB3抑制剂PFK15对胶质瘤H4细胞增殖、迁移、克隆形成和体内成瘤的影响。方法:采集2015年2月1日至2016年1月31日安康市中医医院神经外科手术切除的31例恶性脑胶质瘤及相应瘤旁组织标本,应用免疫组化技术和Western blotting检测胶质瘤和瘤旁组织中PFKFB3的表达水平。用不同浓度的(1.25、2.5、5.0μmol/L)PFK15抑制胶质瘤H4细胞的PFKFB3表达,通过MTT法、EdU细胞增殖实验、细胞划痕实验、Transwell侵袭实验、克隆形成实验和裸鼠体内成瘤实验分别观察PFK15对H4细胞增殖、迁移、侵袭、克隆形成和体内成瘤的影响。结果:PFKFB3在胶质瘤组织中阳性率显著高于瘤旁组织[(80.60±8.98)%vs(41.57±10.16)%,P<0.05]。MTT和EdU实验结果显示,PFK15可明显抑制H4的增殖活性(P<0.05),且抑制作用具有浓度依赖性。PFK15处理后的H4细胞迁移、侵袭和克隆形成能力明显低于对照组(均P<0.05)。注射PFK15的裸鼠H4细胞移植瘤体积明显小于对照组裸鼠[(254.15±154.25)vs(801.52±224.25)mm3,P<0.01]。结论:PFKFB3在恶性胶质瘤组织中高表达,下调PFKB3可显著抑制胶质瘤H4细胞的恶性生物学行为和体内成瘤,PFKFB3可作为恶性胶质瘤生物治疗潜在的分子靶点。
Objective： To evaluate the expression of 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3（PFKFB3） in malignant glioma tissues and the effects of inhibitor of PFKFB3（PFK15） on the proliferation, migration, invasion, clone formation and tumorigenesis of H4 cells. Methods： Malignant brain glioma tissues and corresponding paratumor tissues from 31 patients, who were hospitalized in Department of Neurosurgery, Ankang Hospital of Traditional Chinese Medicine during February 1, 2015 to January 31, 2016 for operative treatment,were collected for this study. Immunohistochemistry and western blotting assays were applied to detect the expression of PFKFB3 in collected tissues. PFKFB3 in H4 cells were blocked by PFK15（1.25, 2.5, 5.0 μmol/L）. The effect of PFK15 on proliferation, migration, clone formation and tumorigenesis of H4 cells were determined by MTT assay, Ed U incorporation assay, wound healing assay, Transwell assay, colone formation assay and in vivo xenograft bearing nude mice model respectively. Results： Positive expression rate of PFKFB3 was significantly higher in malignant glioma tissues compared with normal adjacent tissues[（80.60±8.98）% vs（41.57±10.16）%, P〈0.05]. The results of MTT assay and Ed U incorporation assay indicated that PEK15 significantly inhibited the proliferation of H4 cells in a concentration dependent manner. The migration, invasion and clone formation activity of H4 cells were significantly reduced by treatment with PFK15（all P〈0.05）.In tumor bearing nude mice, the tumor volume of mice treated with PFK15 was significantly smaller than that of mice from control group（[254.15±154.25] vs [801.52±224.25] mm3, P〈0.05）. Conclusion： PFKFB3 was highly expressed in malignant glioma tissues. Blocking of PFKFB3 by PFK15 significantly reduced the malignant biological behaviors and tumorigenesis of H4 cells in vitro and in vivo, which may serve as a promising target for the treatment of malignant gliomas.
作者
陈向荣
杜菊梅
吴宗涛
CHEN Xiangrong;DU Jumei;WU Zongtao(Department of Neurosurgery, Ankang Hospital of Traditional Chinese Medicine, Ank- ang 725000, Shaanxi, China;Department of Encephalopathy, the Second Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine, Xianyang 712000, Shaanxi, China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2018年第4期363-369,共7页
Chinese Journal of Cancer Biotherapy
基金
陕西中医药大学科研基金资助项目(No.2016QN22)~~
