液氮冻存瘤块构建VX2肝癌模型效果研究
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摘要
目的观察液氮冻存的活性良好VX2兔肝癌瘤块复苏后构建兔肝癌模型的效果,并与常规方法构建兔肝癌模型作比较。方法选取生长状态良好的VX2瘤块,剔除坏死组织并用锡箔纸包裹,液氮冻存3个月。20只日本大耳白兔随机分为两组,A组(对照组,n=10)予新鲜瘤块肝脏原位种植法构建兔肝癌模型,B组(实验组,n=10)予新鲜瘤块液氮冻存3个月复苏后肝脏原位种植法构建兔肝癌模型。2周后作增强CT扫描和DSA血管造影观察成瘤效果及瘤体血供情况。荷瘤兔处死后观察大体标本并作苏木精-伊红(HE)染色、血管内皮细胞生长因子(VEGF)和CD31免疫荧光染色,后者用于计算微血管密度(MVD)。结果 A组和B组建模成功率均为100%。两组肿瘤生长、VEGF和MVD表达均无明显差异。结论液氮冻存瘤块构建的兔VX2肝癌模型CT、DSA影像学表现及组织学特征与常规方法无显著差异,瘤块整体冻存法可较好地保存瘤株活性,从而节省人力物力。
Objective To discuss the establishment of rabbit VX2 hepatocellular carcinoma(HCC model by using tumor lump which is frozen in liquid nitrogen and has good activity after resuscitation, and to compare the modeling effect with that by using conventional method. Methods The VX2 tumor blocks with good growth state were selected, the necrotic tissues were remove and the tissues in good condition were wrapped with tinfoil and frozen in liquid nitrogen for three months. Twenty Japanese white rabbits were randomly divided into two groups: group A(control group, n=10) and group B(experiment group, n=10). In group A, rabbit liver cancer model was established by liver transplantation of fresh tumor lump with in situ planting method. In group B, the fresh liver tumor lumps were frozen in liquid nitrogen for three months first,then, the resuscitated tumor tissues were implanted into the liver with in situ planting method. Two weeks later, enhanced CT scan and DSA angiography were performed to check the tumor-modeling effect and tumor blood supply. After tumor-loaded rabbits were sacrificed, the gross specimens were removed and sent for pathological examinations. HE staining was adopted, and immunofluorescence staining of vascular endothelial growth factor(VEGF) and CD31 was used to calculate the microvessel density(MVD). Results In both group A and group B, the success rate of modeling was 100%. No significant differences in tumor growth rate and the expression levels of VEGF and MVD existed between the two groups. Conclusion The imaging manifestations on CT and DSA and the histologic characteristics of rabbit VX2 HCC model, which is established with tumor lump frozen in liquid nitrogen, are no obviously different from those of rabbit VX2 HCC model, which is established with conventional modeling methods. The freezing method of whole tumor lump can well preserve the tumor strain activity, thereby saving the manpower and material resources in the preparation of VX2 HCC model.
Objective To discuss the establishment of rabbit VX2 hepatocellular carcinoma(HCC model by using tumor lump which is frozen in liquid nitrogen and has good activity after resuscitation, and to compare the modeling effect with that by using conventional method. Methods The VX2 tumor blocks with good growth state were selected, the necrotic tissues were remove and the tissues in good condition were wrapped with tinfoil and frozen in liquid nitrogen for three months. Twenty Japanese white rabbits were randomly divided into two groups: group A(control group, n=10) and group B(experiment group, n=10). In group A, rabbit liver cancer model was established by liver transplantation of fresh tumor lump with in situ planting method. In group B, the fresh liver tumor lumps were frozen in liquid nitrogen for three months first,then, the resuscitated tumor tissues were implanted into the liver with in situ planting method. Two weeks later, enhanced CT scan and DSA angiography were performed to check the tumor-modeling effect and tumor blood supply. After tumor-loaded rabbits were sacrificed, the gross specimens were removed and sent for pathological examinations. HE staining was adopted, and immunofluorescence staining of vascular endothelial growth factor(VEGF) and CD31 was used to calculate the microvessel density(MVD). Results In both group A and group B, the success rate of modeling was 100%. No significant differences in tumor growth rate and the expression levels of VEGF and MVD existed between the two groups. Conclusion The imaging manifestations on CT and DSA and the histologic characteristics of rabbit VX2 HCC model, which is established with tumor lump frozen in liquid nitrogen, are no obviously different from those of rabbit VX2 HCC model, which is established with conventional modeling methods. The freezing method of whole tumor lump can well preserve the tumor strain activity, thereby saving the manpower and material resources in the preparation of VX2 HCC model.
引文
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[11]钱亭,陈茂振,高峰,等.两种方法建立兔VX2肝癌模型的比较及影像学评估[J].介入放射学杂志,2014,23:58-61.
[12]金光鑫,王军,仇晓霞,等.MR导引经皮穿刺瘤块种植法构建兔VX2肝癌模型[J].介入放射学杂志,2016,25:980-983.
[2]Xu JY,Yang Y,Xie R,et al.The NCX1/TRPC6 complex mediates TGF beta-driven migration and invasion of human hepatocellular carcinoma cells[J].Cancer Res,2018,78:2564-2576.
[3]Omyla-Staszewska J,Deptala A.Effective therapeutic management of hepatocellular carcinoma:on the basis of a clinical case[J].Contemp Oncol(Pozn),2012,16:60-63.
[4]赵倩,颜志平.载药微球经导管动脉化疗栓塞治疗肝癌研究进展[J].介入放射学杂志,2017,26:1052-1056.
[5]Gnutzmann D,Kortes N,Sumkauskaite M,et al.Transvascular therapy of hepatocellular carcinoma(HCC),status and developments[J].Minim Invasive Ther Allied Technol,2018,27:69-80.
[6]管阳,刘凤永,付金鑫,等.肝癌动物模型与介入实验操作应用[J].介入放射学杂志,2017,26:1046-1051.
[7]江雄鹰,罗荣光,黄金华,等.兔VX2肝癌模型建立与经兔股动脉微导管超选择性肝左动脉插管技术的探讨[J].介入放射学杂志,2011,20:214-217.
[8]李智,倪才方,董凤林,等.兔VX2肝癌模型的建立及其生长转移特性的观察[J].介入放射学杂志,2009,18:691-694.
[9]陈胜利,全毅,黄子诚,等.Lp-THAE诱导兔VX2肝癌细胞凋亡[J].介入放射学杂志,2007,16:406-410.
[10]秦建民,李琦,朱慧蓉.肝癌的药物靶向治疗[M].郑州:郑州大学出版社,2015:140-144.
[11]钱亭,陈茂振,高峰,等.两种方法建立兔VX2肝癌模型的比较及影像学评估[J].介入放射学杂志,2014,23:58-61.
[12]金光鑫,王军,仇晓霞,等.MR导引经皮穿刺瘤块种植法构建兔VX2肝癌模型[J].介入放射学杂志,2016,25:980-983.