摘要
目的探讨不同剂量分割放射治疗对直肠癌肿瘤内血管生成的影响及其潜在机制。方法收集2017-2020年重庆医科大学附属第一医院肿瘤科收治的56例直肠癌患者病理学资料进行统计学分析,采用免疫组化检测大分割放疗(5 Gy×5次,1周内完成)和常规分割放疗(2 Gy×25次,5周内完成)后直肠癌患者癌组织中微血管密度,并使用Western blot检测癌组织中的低氧诱导因子-1α(hypoxia inducible factor 1-alpha,HIF-1α)和血管内皮生长因子(vascular endothelial growth factor,VEGF)蛋白表达水平。将培养的直肠癌细胞及人脐静脉内皮细胞分为0、4、12、24 Gy剂量辐照组,采用CCK-8法检测两种细胞各剂量辐照组的增殖情况,Western blot检测各剂量辐照组直肠癌细胞中VEGF和HIF-1α蛋白表达水平,ELISA检测直肠癌细胞上清液中VEGF的浓度。收集各剂量辐照组的直肠癌细胞条件培养基,干预培养并用CCK-8法和克隆形成实验观测人脐静脉内皮细胞的增殖情况。结果常规分割放疗组和大分割放疗组在病理肿瘤退缩分级(tumor regression grade,TRG)上差异无统计学意义,直肠癌患者术后癌组织免疫组化显示,大分割放疗组微血管密度计数(microvessel density,MVD,32.00±3.61)显著高于常规分割放疗组(22.33±3.06,P<0.05);Western blot结果显示,大分割放疗组癌组织内HIF-1α和VEGF蛋白表达水平显著高于常规分割放疗组(P<0.01)。细胞实验结果显示,不同剂量放射照射后,呈剂量依赖性降低直肠癌细胞和血管内皮细胞的增殖能力(P<0.05)。直肠癌细胞中VEGF和HIF-1α的蛋白表达水平和直肠癌细胞上清液中VEGF的蛋白浓度随着辐照剂量的增加逐渐升高(P<0.01)。大剂量辐照后直肠癌细胞培养基干预人脐静脉内皮细胞能显著提高其增殖能力(P<0.01)。结论大剂量放疗能抑制血管内皮细胞增殖,同时可能通过促进直肠癌细胞HIF-1α和VEGF高表达,导致肿瘤血管代偿性生成增�
Objective To explore the effects of different dose fractionated radiotherapy on angiogenesis in rectal cancer and its potential mechanism.Methods The pathological data of 56 rectal cancer patients admitted to our hospital from 2017 to 2020 were collected for statistical analysis.The microvessel density(MVD)in the rectal cancer tissues under hypofractionated radiotherapy(HFR)and conventional fractionated radiotherapy(CFR)were respectively detected by immunohistochemistry.The expression levels of hypoxia inducible factor 1-alpha(HIF-1α)and vascular endothelial growth factor(VEGF)in the rectal cancer tissues were determined by Western blotting.Then the cultured rectal cancer SW1463 cells and human umbilical vein endothelial cells(HUVECs)were irradiated at doses of 0,4,12 and 24 Gy,respectively.CCK-8 assay was subsequently used to test the proliferation of both cells in each group,Western blotting was performed to measure the levels of VEGF and HIF-1αin SW1463 cells,and ELISA assay was adopted to detect the concentration of VEGF in the supernatant of rectal cancer cells.In addition,CCK-8 assay and colony formation assay were carried out to observe the proliferation of HUVECs after intervened with different doses of irradiated cultured medium.Results Chi-square test showed no significant difference in pathological tumor regression grade(TRG)between the HFR and CFR groups.The results of immunohistochemistry indicated that the MVD in the HFR group(32.00±3.61)was significantly higher than that in the CFR group(22.33±3.06,P<0.05).Western blotting also revealed the protein levels of HIF-1αand VEGF were remarkably up-regulated in the HFR group(P<0.01).In in vitro study,high-dose radiation dose-dependently reduced the proliferation of SW1463 cells and HUVECs(P<0.05).However,the expression levels of VEGF and HIF-1αin rectal cancer cells and the content of VEGF in the supernatant were increased gradually with the rise of irradiation dose(P<0.01).Correspondingly,the proliferation of HUVEC cells was prominently enhanced by
作者
童潮旭
张涛
龚惠英
毛伟灵
李龙浩
TONG Chaoxu;ZHANG Tao;GONG Huiying;MAO Weiling;LI Longhao(Department of Oncology,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400016,China)
出处
《陆军军医大学学报》
CAS
CSCD
北大核心
2022年第19期1979-1987,共9页
Journal of Army Medical University
基金
重庆市科卫联合医学科研项目(2021MSXM318)
