人胰腺混合性导管-内分泌癌细胞系的建立

The establishment of a human pancreatic mixed ductal endocrine carcinoma cell line

目的 建立胰腺混合性导管内分泌癌细胞系。方法 从一男性患者胰头切除的肿瘤组织取材，切割为细小的组织块接种于裸鼠皮下，经过裸鼠的两次过渡后，再取第二次裸鼠皮下的肿瘤组织，切割为非常小的组织块，用含有１０ ％ 胎牛血清的培养基接种至培养瓶进行体外培养。结果该细胞系目前已传至５５ 代。这一细胞系（ＰＣＥＮ）同时具有内、外分泌胰腺癌的形态特征，并得到了免疫组化标记和组织化学染色的证实。电镜下，部分细胞内含有神经分泌颗粒。生长特性表现为群体倍增时间相对较长。染色体核型说明ＰＣＥＮ有两个细胞群，一个细胞群的染色体众数为４８ 条，另一个的为５６ 条。癌基因Ｋｉｒａｓ１２ 密码子突变。应用放免法在ＰＣＥＮ 细胞匀浆及培养上清液中均检测到多种胃肠激素和癌胚抗原。结论 ＰＣＥＮ细胞系的细胞、分子生物学的特点充分说明，该细胞系不仅同时具有内、外分泌胰腺的形态特征，还具有其独特的生长特性、染色体核型及癌基因的变化，更重要的是ＰＣＥＮ同时具有内、外分泌胰腺的功能。ＰＣＥＮ的建立，为深入研究内、外分泌胰腺肿瘤的各种细胞和分子生物学特性提供了一个较有价值的体外模型。

Objective Establishment of a human pancreatic mixed ductal endocrine carcinoma cell line. Methods A specimen of this type of cancer was obtained from a male patient at the head of pancreas. The tumor tissue was minced into pieces and inoculated subcutaneously in nude mice. With two successive subcutaneous implants, a piece of tumor tissue taken from the 2 nd implanted neoplasm of a nude mouse was minced into minute tissue masses and inoculated in a media containing 10% fetal bovine serum. Results Currently, it has been passed to the 55 th passage. Cells of this cell line (PC EN) kept simultaneously the morphologic patterns of a pancreatic ductal and endocrine tumor which were verified by histochemistry and immunohistochemistry assays. Some of the PC EN cells showed presence of neuroendocrine granules by electron microscopy. The growth rate of this cell line was relatively low. Kakryotype analysis showed two cell populations with its major mode of chromosomes numbers as 48 in one, and 56 in the other. Study on K ras gene demonstrated that PC EN harbored a mutated codon 12. Several gastrointestinal hormones and CEA were detected in PC EN cells and in the supra cultured medium using radioimmunoassay. Conclusions The cellular and molecular biologic characteristics of PC EN cells illustrated that this cell line preserved the morphology of a pancreatic ductal and endocrine tumor and possessed a special growth property, chromosome karyotype, and mutation of Ki ras gene, but also showed a biologic exocrine and endocrine functions of the pancreas. The establishment of PC EN cell line will provide a useful model in vitro for further investigating the cellular and molecular biology of exocrine and endocrine tumors of the pancreas.