不同剂量N-亚硝基双-2-氧丙基诱导仓鼠胰腺癌的成模效果及对相关基因表达的影响

Effects of different doses of BOP on induced hamster model of pancreatic cancer and related gene expression

目的 探讨不同时间、不同剂量N-亚硝基双-2-氧丙基（N-nitrosobis-2-oxopropylamin,BOP）诱导仓鼠胰腺癌的成模效果及仓鼠胰腺癌组织COX-2、5-LOX、GAL-1、GAL-3、MHC-Ⅰ、MHC-Ⅱ等基因表达的变化.方法 72只仓鼠按完全随机法分为3组,每组24只.A组采用小剂量BOP长周期皮下注射诱导造模（10 mg/kg体质量,每周1次,连续8周）;B组采用大剂量BOP短周期皮下注射诱导造模（首次剂量70 mg/kg体质量,后3周每周20 mg/kg体质量）;C组为未注射BOP的对照组.成模后取胰腺组织及肝脏组织行病理组织学检查.采用实时PCR法及蛋白质印迹法检测仓鼠胰腺癌组织COX-2、5-LOX、GAL-1、GAL-3、MHC-Ⅰ、MHC-Ⅱ等基因的表达.结果 A组仓鼠中19只成功诱导胰腺癌模型;B组均未见胰腺肿瘤形成,而18只仓鼠成功诱导肝内胆管细胞癌;C组胰腺及肝脏均未见异常.仓鼠胰腺癌组织COX-2、5-LOX、GAL-1、GAL-3、MHC-Ⅰ mRNA的表达量分别为对照组的1.93、2.03、1.57、5.55和3.54倍,差异有统计学意义（P值均＜0.05）;COX-2、5-LOX、GAL-1、GAL-3、MHC-Ⅰ蛋白的表达量分别为2.47 ±0.22、0.56 ±0.05、1.23 ±0.05、1.55 ±0.13、2.02 ±0.01,而对照组仓鼠正常胰腺组织的蛋白表达量分别为0.80±0.08、0.39 ±0.01、0.60 ±0.01、0.53 ±0.03、0.40 ±0.02,两组差异均有统计学意义（P值均＜0.05）.两组MHC-ⅡmRNA及蛋白表达的差异无统计学意义.结论 小剂量BOP长周期皮下注射能成功诱导仓鼠胰腺癌模型,胰腺癌组织的COX-2、5-LOX、GAL-1、GAL-3、MHC-Ⅰ基因表达上调,可能参与仓鼠胰腺癌的发生和发展.

Objective To investigate the effects of different timing and doses of BOP on the construction model of hamster pancreatic cancer,and on the expression of COX-2,5-LOX,GAL-1,GAL-3,MHC-Ⅰ,MHC-Ⅱ gene.Methods Seventy-two hamsters were randomly divided into 3 groups,hamsters in A group were treated by subcutaneous injection of small dose long cycle of BOP （10 mg/kg,once a week for 8 weeks）,hamsters in B group were treated with large doses of BOP and short period of subcutaneous injection （initial dose 70 mg/kg,then 20 mg/kg,once a week for 3 weeks）,hamsters in C groups were control group.The hamsters were sacrificed after the model was successfully established and pancreas tissue,liver tissue were harvested for pathological examinations.Real-time PCR and Western-blot were used to detect the expression of COX-2,5-LOX,GAL-1,GAL-3,MHC-Ⅰ,MHC-Ⅱ gene in pancreatic cancer tissue.Results Pancreatic cancer model was successfully established in 19 out of 24 hamsters in A group,while 18 hamsters in B group developed cholangiocarcinoma but not pancreatic cancer.Hamsters in C groups remained healthy.The expressions of COX-2,5-LOX,GAL-1,GAL-3,MHC-Ⅰ mRNA in pancreatic cancer tissue were 1.93,2.03,1.57,5.55 and 3.54 folds higher than that in control group,and the difference between the two groups was statistically significant （P ＜ 0.05）.The expressions of COX-2,5-LOX,GAL-1,GAL-3,MHC-Ⅰ protein were 2.47 ±0.22,0.56 ±0.05,1.23 ±0.05,1.55 ±0.13 and 2.02 ±0.01,while the corresponding values in control group were 0.80 ± 0.08,0.39 ± 0.01,0.60 ± 0.01,0.53 ± 0.03 and 0.40 ± 0.02,and the difference between the two groups was statistically significant （P ＜ 0.05）.The expressions of MHC-Ⅰ mRNA and protein between the two groups were not statistically significant.Conclusions Small dose,long cycle subcutaneous injection of BOP can successfully induce pancreatic cancer mode in hamster.The expressions of COX-2,5-LOX,GAL-1,GAL-3,MHC-Ⅰ are up-regulated,which may be involved in the development of pancreatic cancer.