期刊：
Journal of Zhejiang University. Science. B,2009年10(3):172-179 ISSN：1673-1581
通讯作者：
He, Ying-chun
作者机构：
[Tian, Dao-fa; He, Ying-chun] Chinese Med Univ Hunan, Fac Integrat Med, Changsha 410007, Peoples R China.;[Lu, Fang-guo] Chinese Med Univ Hunan, Fac Basic Med, Changsha 410007, Peoples R China.;[Tang, Fa-qing] Cent S Univ, Dept Clin Lab, Xiangya Hosp 1, Changsha 410008, Peoples R China.
通讯机构：
[He, Ying-chun] C;Chinese Med Univ Hunan, Fac Integrat Med, Changsha 410007, Peoples R China.
摘要：
OBJECTIVE: To investigate the enhancive effect of N,N'-dinitrosopiperazine (DNP) on induced carcinogenesis in nasal and/or nasopharyngeal epithelia among TgN(p53mt-LMP1)/HT transgenic mice to examine the underlying mechanism for the development of nasopharyngeal carcinoma (NPC). METHODS: TgN(p53mt-LMP1)/HT transgenic mice and the same strain of C(57)BL/6J wild-type mice both at the age of 5 months were randomly divided into 2 groups in parallel, respectively, i.e., TgN(p53mt-LMP1)/HT cancerous lesion-inducing group (TI), TgN(p53mt-LMP1)/HT control group (TC), C57BL/6J cancerous lesion-inducing group (CI), and C57BL/6J control group (CC). TI and CI mice were treated only with DNP for 16 weeks, twice each week, while TC and CC mice were given the same volume of saline as controls. At the end of treatment, animals were sacrificed to collect epithelial tissue samples from nasal cavity and nasopharynx for pathohistological evaluation by haematoxylin and eosin (HE) staining and for determination on the expression of TRAF2, c-Jun, and p16 by immunohistochemistry. RESULTS: Atypical hyperplasia was more significant in the samples of TI than in those of TC, CI, and CC, with the rates of lesions being 90%, 10%, 0, and 0 (P<0.01) respectively, though DNP was used alone in a much shortened inducing period at less dosage and without the use of carcinogenic promoter 12-O-tetradecanoylphorbol-13-acetate as usual. The expressions of tumor necrosis factor (TNF) receptor-associated factor 2 (TRAF2) and c-Jun in these samples were significantly up-regulated in TI (P<0.01), while the expression of p16 was significantly lower in TI than in the other groups (P<0.01). CONCLUSION: TgN(p53mt-LMP1)/HT mice hold inherited constitutional defect in immune surveillance function, which can be aggravated by environmental carcinogens, such as DNP used even though in a much less strength. The enhanced carcinogenesis-inducing effect of DNP on TgN(p53mt-LMP1)/HT mice should be closely associated with abnormal signaling of activator protein-1 (AP-1) pathway, especially up-regulated expressions of TRAF2 and c-Jun, and down-regulated expression of p16.
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20R,随机分为2组:①TgN(p53mt-LMP1)/HT诱癌中药组[中药组,10R,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,10R,以15 ml/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15 ml/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WesternEO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(JP<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±,10 VS 132±10,鼻咽:155±12 VS 133±10,P值均<0.01),c-jun基因的表达水平显著下调(鼻腔:111±9 VS 161±10;鼻咽:1104±10 VS 159±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12±12,c-Jun分别为164±13和164±9)的差异无显著性(Jp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.
作者机构：
[Tian, D.; Jiang, J.; Lu, F.; He, A.; He, Y.; Liu, G.] Department of Integrative Medicine, Taditional Chinese Medicine University of Hu'nan, Changsha 410007, China
通讯机构：
[He, Y.] D;Department of Integrative Medicine, Taditional Chinese Medicine University of Hu'nan, China
作者机构：
[Tian, D. F.; Liu, S. J.; He, Y. C.] Tradit Chinese Med Univ Hunan, Fac Integrat Med, Changsha 410007, Peoples R China.;[Ling, C. Q.; Liu, S. J.] Changhai Hosp, Dept Tradit Chinese Med, Shanghai, Peoples R China.;[Sun, Y. M.] Chinese Acad Sci, Inst Genet & Dev Biol, Beijing, Peoples R China.;[Zeng, L.] Huan Provincial Tumor Hosp, Dept Pathol, Changsha, Peoples R China.;[Sun, S. H.; He, Y.] Second Mil Med Univ, Dept Med Genet, Shanghai, Peoples R China.
通讯机构：
[Tian, D. F.] T;Tradit Chinese Med Univ Hunan, Fac Integrat Med, 113 Shaoshan Middle Rd, Changsha 410007, Peoples R China.
摘要：
Because the focus of nasopharyngeal carcinoma (NPC) is very close to intracranial organs, it often makes incursions into cranial cavity. Identification of intracranial invasion-associated indicators will provide potential therapeutic targets for NPC patients with intracranial invasion. In this regard, Human Xpro(TM) HC-plus cancer-related gene chip was utilised to screen intracranial invasion-associated genes for NPC from the biopsied primary focus tissue samples. In all, 8 upregulated and 23 downregulated genes were obtained. VEGF165 and MMP-9, the two upregulated genes, and NM23-H1, the downregulated one, were further confirmed by immunohistochemistry, quantitative real-time PCR and western blot. Invasion-associated cellular and nude mouse models were subsequently employed to study the biological properties of NM23-H1. NM23-H1 expression was significantly lower in 5-8F cells compared with that in 6-10B cells. Moreover, patch-clamp and transwell chamber were adopted to investigate the invading potential-associated biological dynamic mechanisms in the two cell lines, and Ca2+ current and motility were significantly elevated in 5-8F cells compared with that in 6-10B cells. Berberine, an inhibitor of Ca2+ current, could substantially increase the expression of NM23-H1 and decrease 5-8F cell motility. The specificity of berberine on NM23-H1 and cell motility was confirmed by RNAi assay.
