胰腺癌血管内皮细胞生物学特性的实验研究

Study on the biological characteristics of pancreatic cancer vascular endothelial cells

目的研究胰腺癌血管内皮细胞的形态学特征、种属来源、遗传学特征、体外血管形成能力、增殖能力等生物学特征。方法将人胰腺癌肿瘤细胞接种于裸鼠胰腺内，获取胰腺癌血管内皮细胞。体外培养胰腺癌血管内皮细胞，记录传代次数和传代速度，光学显微镜下观察每代细胞的形态学特征。用染色体核型分析技术研究胰腺癌血管内皮细胞的种属来源及遗传学特征，经小管形成实验观察胰腺癌血管内皮细胞的体外血管生成能力。以人脐静脉血管内皮细胞为对照，采用MTT比色法定量分析胰腺癌血管内皮细胞的体外增殖状况。对数据进行单因素方差分析及两两比较。结果在适合的培养条件下，胰腺癌血管内皮细胞每2至3d传代i次，达融合状态时呈单层生长，呈现“铺路石”样特征。胰腺癌血管内皮细胞的种属类型为人，可见大量多倍体细胞、非整数倍染色体细胞、核内染色体缺失、错位及无法分析的片段。胰腺癌血管内皮细胞能在体外形成中空的管状结构。在体外培养48和72h后，胰腺癌血管内皮细胞组的吸光度分别为0．581±0．014和1．082±0．033，显著高于人脐静脉血管内皮细胞组[O.379±0.038（t=8．720，P=0．001）和0．604±0．026（t=19．883，P〈O．01）]。结论胰腺癌血管内皮细胞的种属来源与人胰腺癌细胞相同，具有血管内皮细胞的典型形态特征和体外血管生成能力，具有遗传不稳定性和活跃的增殖能力。

Objective To explore the biological characteristics of pancreatic cancer vascular endothelial cells, including the aspects of morphology, species, genetics, vascular formation ability, and proliferation ability in vitro. Methods The human pancreatic cancer cells were inoculated in nude mice pancreas to get pancreatic cancer vascular endothlial cell. The pancreatic cancer vascular endothelial cells were cultured in vitro. The passage number and passage time were recorded. The morphological features under common microscopy of each passage were observed. The species origin and genetic characteristics of the pancreatic cancer vascular endothelial cells were detected by karyotype assay. The ability of angiogenesls of the pancreatic cancer vascular endothelial cells in vitro was determined by tube formation assay. The proliferation of the pancreatic cancer vascular endothelial cells in vitro was measured by MTT method. The data were analyzed by one way analysis of variance and paired difference test. Results Under appropriate culture condition, the pancreatic cancer endothelial cells were passaged every two to three days. Once confluence was attained, the cells were in monolayer growth and with cobblestone feature. The species type of the pancreatic cancer vascular endothelial cells was human type. A large number of polyploid ceils, non-integer multiple chromosomes cells, nuclear chromosome loss, nuclear chromosome dislocation, and unanalyzahle fragments were observed. The pancreatic cancer vascular endothelial cells could form a hollow tubular structure in vitro. After cultured for 48 and 72 hours, the absorbance of the pancreatic cancer vascular endothelial ceils was 0. 581 ±0. 014 and 1. 082 ± 0. 033 respectively, both were significantly higher than those of human umbilical vein vascular endothelial cells （0,379±0. 038, t=8. 720, P=0. 001; 0. 604±0. 026, t= 19. 883, P〈0.01）. Conclusions The species origin of pancreatic cancer vascular endothelial cells is same as human pancreatic cancer cells. The cells have typical morphological features and in vitro angiogenesis formation ability of vascular endothelial cells, whose genetic feature is instable and proliferation is active.