Analysis of oncogenesis in pancreatic cancer using ductal cells tranfered with papilloma virus E6/E7

使用乳头瘤病毒 E6/E7 转染的导管细胞分析胰腺癌的肿瘤发生

• 批准号: 11670481
• 负责人: KAWABE Takao
• 金额: $ 2.3万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670481/
• 关键词: Papilloma virus; Gene transfer; Pancreatic duct cell; Pancreatic cancer; パピローマウィルス; 膵

1. Monolayer culture of pancreatic duct cells. We established the monolayer culture of rat pancreatic duct cells. Then we also tried to culture of human pancreatic ductal cells, but it was not successful. We did not obtain a large amount of human materials and the materials were not brought to us soon after they were obtained from the patients. These were considered part of the reasons for our failure in human pancreatic duct cell culture.2. Construction of recombinant adenovirus vectors. Bcl-2 and bcl-XL are considered to be anti-apoptotic genes and mutated k-ras to promote cell growth. We tried to construct adenovirus vectors integrated with these genes using 293 cells by COS-TCP method.The expression of Bcl-XL was confirmed by Western blotting in the cells infected with recombinant adenovirus. An elevation of the bio-activity was also revealed by the anti-apoptotic effects of Bcl-XL, which was accessed by resistant property to Cisplatin.The clones of the recombinant virus integrated with bcl-2 or mutated k-ras were obtained. However, the amount of the virus titer was not enough for further experiments in 293 cells. To obtain enough amount, another technique, such as CRE-loxP or conditional expression system, may be needed in further investigations.

1.胰管细胞的单层培养。我们建立了大鼠胰管细胞的单层培养。随后我们也尝试培养人胰腺导管细胞，但没有成功。我们没有获得大量的人体材料，而且从病人那里获得材料后也没有很快送到我们这里。这些被认为是我们人胰管细胞培养失败的部分原因。2．重组腺病毒载体的构建。 Bcl-2和bcl-XL被认为是抗凋亡基因，突变的k-ras可促进细胞生长。我们尝试通过COS-TCP方法利用293细胞构建整合这些基因的腺病毒载体。通过Western blotting证实重组腺病毒感染的细胞中Bcl-XL的表达。通过对顺铂的抗性，Bcl-XL的抗凋亡作用也显示出生物活性的提高。获得了整合有bcl-2或突变的k-ras的重组病毒的克隆。然而，病毒滴度不足以在293细胞中进行进一步实验。为了获得足够的量，进一步研究可能需要另一种技术，例如 CRE-loxP 或条件表达系统。

项目成果

Maeda S, Yoshida H, Ogura K, Yamaji Y, Ikenoue T, Mitsushima T, Tagawa H, Kawaguchi R, Mori K, Mafune K, Kawabe T, Shiratori Y, Omata M.: "Assessment of gastric carcinoma risk associated with Helicobacter pylori may vary depending on the antigen used : Ca

Maeda S、Yoshida H、Ogura K、Yamaji Y、Ikenoue T、Mitsushima T、Takawa H、Kawaguchi R、Mori K、Mafune K、Kawabe T、Shiratori Y、Omata M.：“与幽门螺杆菌相关的胃癌风险评估

Ikenoue T, Togo g, Kawabe T, et al.: "Frameshift mutation at mononucleotide repeats in RAD50 recombinational DNA rep air gene in colorectal cancer with microsatellite Instability"Jpn J Cancer Res. 92・6. 587-591 (2001)

Ikenoue T、Togo g、Kawabe T 等：“具有微卫星不稳定性的结直肠癌中 RAD50 重组 DNA 修复基因中单核苷酸重复的移码突变”Jpn J Cancer Res 92·6。

Obi S, Shiratori Y, Kawabe T, et al.: "Early detection of haemobilia associated with percutaneous ethanol injection for hepatocellular carcinoma"Eur. J. Gastroenterol. Hepatol. 12・3. 285-290 (2000)

Obi S、Shiratori Y、Kawabe T 等人：“肝细胞癌经皮乙醇注射相关的出血的早期检测”Eur. Gastroenterol 12·3 (2000)。

Shiratori Y,Kanai F,Ohashi M, et al: "Strategy of liver-directed gene therapy : present status and future prospects"Liver. 19・4. 265-74 (1999)

Shiratori Y、Kanai F、Ohashi M 等：“肝脏定向基因治疗策略：现状和未来前景” 肝脏 265-74 (1999)。

Yamagata M, Kawabe T, Tada M, et al.: "In vitro sonographic evaluation of common bile duct stones and fragments with a high-frequency microprobe"J. Clin. Ultrasound. 27・5. 249-257 (1999)

Yamagata M、Kawabe T、Tada M 等人：“利用高频微探针对胆总管结石和碎片进行体外超声评估”J. Clin 27・5 (1999)。