三种原位种植人胰腺癌裸鼠模型的建立及其血清代谢组学

Establishment of three human pancreatic cancer orthotopic xenograft nude mice models and serum metabolomics

目的应用代谢组学的方法，分析三种不同分化程度人胰腺癌细胞株原位种植胰腺癌裸鼠模型血清与正常裸鼠血清之间的代谢差异。方法分别将三种人胰腺癌细胞株SW1990（中分化）、BxPC-3（低一中分化）、Panc-1（低分化）接种于裸鼠皮下。皮下成瘤后，取皮下癌组织原位移植于裸鼠胰腺，从而建立裸鼠原位种植胰腺癌模型。造模成功后，采集血清标本，然后采用基于核磁共振的代谢组学技术，联合多变量统计分析处理，筛选出三种胰腺癌裸鼠模型与正常裸鼠血清标本的差异代谢物。结果三种裸鼠原位种植癌模型均成功构建。SW1990组成瘤率为79％（11／14），病死率为7％（1／14）；BxPC-3组成瘤率为93％（13／14），无死亡；Panc-1组成瘤率为86％（12／14），病死率为7％（1／14）。三种胰腺癌裸鼠血清中肌酸、丙氨酸、谷氨酰胺、1-甲基组氨酸、异亮氨酸、乳酸、苯丙氨酸、色氨酸及缬氨酸水平均显著高于正常裸鼠，而甘油磷酸胆碱及葡萄糖水平显著低于正常裸鼠。三种胰腺癌裸鼠血清中异亮氨酸及缬氨酸水平随肿瘤分化程度的降低而升高。结论采用原位种植的方法可建立稳定可靠且成瘤率较高的裸鼠胰腺癌模型。胰腺癌裸鼠与正常裸鼠两者糖代谢、脂质代谢及氨基酸代谢存在显著差异。不同人胰腺癌原位种植裸鼠模型血清中具有某些相同的代谢物，可作为诊断胰腺癌的潜在标志物。

Objective To analyze the metabolic profile in serum between normal and orthotopic xenograft nude mice burdened with three human pancreatic cancer cell lines, which were differentiated differ- ently. Methods Human pancreatic cancer lines SW1990, BxPC-3 and Panc-1 were subcutaneously injec- ted into the nude mice, respectively. When the tumor volume reached 1.0 cm3 , the nude mice were eutha- nized and the tumor tissues were removed and implanted to the pancreas to establish the orthotopie xenograft mice model. The serum from three orthotopic xenograft tumor nude mice and the normal controls were col- lected and then analyzed by 1H nuclear magnetic resonance spectroscopy. Results The three orthotopie xenograft nude mice models were successfully established. In SW1990, BxPC-3 and Pane-1 group, the or- thotopie xenograft tumor formation rate was 79% （ 11/14）, 93% （13/14） and 86% （ 12/14）, while the mortality was 7% （1/14）, 0 and 7% （ 1/14）, respectively. Compared with control group, the content of metabolites in the serum of orthotopie xenograft tumor nude mice was increased including ereatine, alanine, glutamine, 1-methylhistidine, isoleucine, lactate, phenylalanine, tryptophan and valine, but the glycerol- phosphocholine （GPC） and glucose levels were reduced. As the tumors progressed to be more malignant, the content of valine and isoleucine tended to increase. Conclusions The establishment of the orthotopie implantation tumor nude mice model was stable and reliable with high tumor formation rate. Obvious meta- bolic differences of glucose, lipid and amino acids were observed between normal and human pancreatic cancer tumor burdening nude mice models. The common metabolic features identified in all three nude micemodels burdened with human pancreatic cancer could be used as the potential markers for diagnosing human pancreatic cancer.