益气解毒方对DNP诱导TgN(p53mt-LMP1)/HT小鼠鼻腔/鼻咽上皮癌前病变及p16和c-jun基因表达的影响  

Interference of Yiqijiedu formula in DNP inducing effects on the occurrence of precancerous lesions of nasal and nasopharyngeal epithelia in TgN(p53mt-LMP1)/HT transgenic mice

作　　者：何迎春[1] 卢芳国[1] 田道法[1] 刘刚[1] 

机构地区：[1]湖南中医药大学中西医结合学院,湖南长沙410007

出　　处：《中国癌症杂志》2008年第10期734-739,共6页China Oncology

基　　金：国家自然科学基金(No:0672738);湖南省杰出青年基金(No:03JJY1006);湖南省科技厅国际合作项目(No:06WK3016);湖南省自然科学基金(No:07JJ6038);湖南省教育厅优秀青年基金(No:06B070)。

摘　　要：背景与目的:有研究表明,益气解毒方中药可通过抑制细胞端粒酶的激活而阻断二亚硝基哌嗪(DNP)诱导大鼠鼻咽癌变和人胚鼻咽上皮细胞转化。本研究探讨益气解毒方对经DNP诱导TgN(p53mt-LMP1)/HT小鼠的鼻腔和鼻咽黏膜上皮增殖及c-jun和p16基因表达的影响,为益气解毒方的疗效和可能作用机制提供新的证据。方法:G4代5月龄TgN(p53mt-LMP1)/HT小鼠20只,随机分为2组:①TgN(p53mt-LMP1)/HT诱癌中药组[中药组,10只,以12.91g/(kg?d)剂量灌胃益气解毒方药液];②TgN(p53mt-LMP1)/HT诱癌0.9%氯化钠溶液对照组(氯化钠溶液对照组,10只,以15ml/kg氯化钠溶液灌胃)。另设阴性对照组(用野生型C57BL/6J小鼠10只,以15ml/kg氯化钠溶液灌胃)。用HE染色法观察各组小鼠鼻腔和鼻咽黏膜上皮组织生物学表型特征,比较癌前病变率;免疫组织化学染色法检测各组小鼠鼻腔和鼻咽黏膜上皮组织c-jun和p16基因的表达水平,并采用Western印迹法验证。结果:阴性对照组、氯化钠溶液对照组和中药组鼻腔和(或)鼻咽黏膜上皮癌前病变率分别为0、90%和10%,差异有显著性(P<0.01)。与氯化钠溶液对照组比较,中药组小鼠鼻腔和鼻咽的p16基因表达水平显著上调(鼻腔:157±10vs132±10,鼻咽:155±12vs133±10,P值均<0.01),c-jun基因的表达水平显著下调(鼻腔:111±9vs161±10;鼻咽:110±10vs159±10,P值均<0.01),趋于正常水平,与阴性对照组(鼻腔和鼻咽的p16表达灰度值分别为130±12和129±12,c-Jun分别为164±13和164±9)的差异无显著性(P值均>0.05)。结论:中药益气解毒方可阻逆DNP对TgN(p53mt-LMP1)/HT小鼠鼻腔和鼻咽黏膜上皮组织癌前病变发生和发展的诱导作用,其干预作用与下调c-jun表达和上调p16基因表达密切相关。Background and purpose：The development of nasopharyngeal carcinoma in rat and transformation of embryo nasophryngeal epithelia induced by N,N＇-dinitrosopiperazine （DNP） were interfered by Yiqijiedu formular medicine via inhibiting telomerase. We investigated the impact of Yiqijiedu formula on precancerous lesion of nasal and nasopharyngeal epithelia in TgN （p53mt-LMP1） / HT mice induced by DNP and on its expression level of c-jun and p16 genes. Methods：The fourth generation TgN （p53mt-LMP1） / HT mice aged 5 months were randomly divided into 2 groups：（1）TgN（p53mt-LMP1）/HT cancerous lesion-inducing group （group A） was treated by normal saline with 15ml/kg, and Chinese traditional medicine treating group （group B） was treated by Yiqijiedu formular drug with 12.91 g/（kg？d）. C57BL/6J wild-type mice were chosen as negative control group （group C） treated by normal saline with 15ml/kg. Epithelial tissue samples were taken from nasal cavity and nasopharyngnx of mice respectively for pathohistological evaluation by H-E staining procedures, and expression level of c-Jun and p16 in these tissue sampleswere determined by immunohistochemical methods and western blot. Results：The occurrence rates of precancerous lesions in nasal and/or nasopharyngeal epithelia in Group C, A and B were 0, 90% and 10% respectively （P〈0.01）. Compared with group B, the expression level of p16 gene was 157=10 vs 132±10 and 155±12 vs 133±10 in nasal epithelia; the expression of c-Jun was 111±9 vs 161=10 in nasal epithelia,and 110±10 vs 159=10 in nasopharyngeal epithelia in group A and group B respectively, there were signifi cantly differences （P〈0.01）. There were no signifi cant differences between group A and B Vs. group C in terms of c-jun and p16 gene expression level. Conclusion： Yiqijiedu formula could prevent the inducing effects of DNP on the occurrence and development of precancerous lesions of nasal/ nasopharyngeal epithelia in TgN （p53mt-LMP1）/HT mice.The effects of Yi

关 键 词：鼻腔上皮 鼻咽上皮 癌前病变 益气解毒方 C-JUN P16 

分 类 号：R73-36[医药卫生—肿瘤]