摘要：
Mutations in the PKHD1 gene result in autosomal recessive polycystic kidney disease (ARPKD) in humans. To determine the molecular mechanism of the cystogenesis in ARPKD, we recently generated a mouse model for ARPKD that carries a targeted mutation in the mouse orthologue of human PKHD1. The homozygous mutant mice display hepatorenal cysts whose phenotypes are similar to those of human ARPKD patients. By littermates of this mouse, we developed two immortalized renal collecting duct cell lines with Pkhd1 and two without. Under nonpermissive culture conditions, the Pkhd1(-/-) renal cells displayed aberrant cell cell contacts and tubulomorphogenesis. The Pkhd1(-/-) cells also showed significantly reduced cell proliferation and elevated apoptosis. To validate this finding in vivo, we examined proliferation and apoptosis in the kidneys of Pkhd1(-/-) mice and their wildtype littermates. Using proliferation (PCNA and Histone-3) and apoptosis (TUNEL and caspase-3) markers, similar results were obtained in the Pkhd1(-/-) kidney tissues as in the cells. To identify the molecular basis of these findings, we analyzed the effect of Pkhd1 loss on multiple putative signaling regulators. We demonstrated that the loss of Pkhd1 disrupts multiple major phosphorylations of focal adhesion kinase (FAK), and these disruptions either inhibit the Ras/C-Raf pathways to suppress MEK/ERK activity and ultimately reduce cell proliferation, or suppress PDK1/AKT to upregulate Bax/caspase-9/caspase-3 and promote apoptosis. Our findings indicate that apoptosis may be a major player in the cyst formation in ARPKD, which may lead to new therapeutic strategies for human ARPKD. (C) 2010 Elsevier Inc. All rights reserved.
作者：
Li Li;He Xiu-Sheng*;Luo Qiao;Zhang Zhi-Wei;Yao Xu-Jiong;...
期刊：
生物化学与生物物理进展,2011年38(3):248-253 ISSN：1000-3282
通讯作者：
He Xiu-Sheng
作者机构：
[Wang Li-Li; Zhang Zhi-Wei; Luo Qiao; Duan Rong; He Xiu-Sheng; Yao Xu-Jiong; Li Chun-Cheng; Li Li; Chen Su-Qiong] Univ S China, Canc Res Inst, Hengyang 421001, Peoples R China.;[Chen Zhu-Chu] Cent S Univ, Canc Res Inst, Changsha 410078, Peoples R China.
通讯机构：
[He Xiu-Sheng] U;Univ S China, Canc Res Inst, Hengyang 421001, Peoples R China.
通讯机构：
Cancer Research Institute, Medical College of University of Southern China, 28 Changsheng West Road, China
关键词：
胃癌;AKT蛋白;p27Kip1蛋白;CyclinE蛋白组织芯片
摘要：
目的:探讨AKT、p27Kip1与Cyclin E表达在胃癌发生发展中的意义.方法:收集2008-01/2010-01湘潭市第一人民医院和南华大学附属南华医院的活检标本和胃癌手术切除标本,包括正常胃黏膜组织14例（胃镜活检标本）、非典型增生胃组织10例（胃镜活检标本）、胃癌组织94例、癌旁组织49例,淋巴结转移胃癌组织35例,制作成组织芯片,每个蜡块设计5×10点阵,阵列的最后一排的最后一孔为空白标记,确定方位顺序.采用免疫组织化学染色,检测胃组织芯片中AKT蛋白、p27Kipl蛋白磷酸化及Cyclin E蛋白表达.方向.结果:与正常胃黏膜、癌旁和非典型增生组织相比,AKT蛋白磷酸化和Cyclin E蛋白在原发癌、淋巴结转移癌中表达升高(AKT:85.1％,85.7％ vs 14.3％,26.5％,30.0％; CyclinE:85.1％,82.9％ vs 14.3％,34.7％,20.0％,均P＜0.01),而p27Kip1蛋白磷酸化水平降低(22.3％,17.1％ vs 71.4％,44.9％,40.0％,P＜0.01)；与正常胃黏膜组织比较,AKT蛋白磷酸化在非典型增生组织中表达上调(P=0.26）.结论:AKT、p27Kip1蛋白磷酸化及Cyclin E蛋白表达与胃癌的发生发展有关,随着AKT蛋白磷酸化及Cyclin E蛋白表达水平的升高,p27kipl蛋白磷酸化水平降低.
作者机构：
[赵其辉] Hunan Environment-Biological Polytechnic, Hengyang Hunan, 421005, China;[邱青朝; 贺修胜] Cancer Research Institute, University of South China, Hengyang Hunan 421001, China;[贺修培] Dept. of Otolaryngology, First People's Hospital of Yunnan Province, Kunming 650032, China;[胡波] Hunan Environment-Biological Polytechnic, Hengyang Hunan, 421005, China, Cancer Research Institute, University of South China, Hengyang Hunan 421001, China
通讯机构：
[Hu, B.] H;Hunan Environment-Biological Polytechnic, Hengyang Hunan, 421005, China
作者机构：
[Zhang Zhi-Wei; Cao Jian-Guo; Li Yan-Lan; Luo Zhao-Yang; He Xiu-Sheng] Univ So China, Coll Med, Canc Res Inst, Hengyang 421001, Peoples R China.;[Zhang Zhi-Wei; Li Yan-Lan; Luo Zhao-Yang; He Xiu-Sheng] Univ So China, Coll Med, Dept Pathol, Hengyang 421001, Peoples R China.;[Zhang Zhi-Wei; Li Yan-Lan; Luo Zhao-Yang; He Xiu-Sheng] Univ Key Lab Canc Cellular & Mol Pathol Hunan Pro, Hengyang 421001, Peoples R China.;[Cao Jian-Guo] Hunan Normal Univ, Coll Med, Dept Pharmacol, Changsha 410013, Hunan, Peoples R China.
通讯机构：
[Zhang Zhi-Wei] U;Univ So China, Coll Med, Canc Res Inst, Hengyang 421001, Peoples R China.
摘要：
Objective： Analysis of improved three site-directed mutagenesis methods in a novel gene recombinants construction. Methods： Overlapping extension PCR, MutanBest kit and Quikchange site-directed mutagenesis kit were used to construct mutants con- raining a single different residue from wild type gene. By Stratagagene primers design on-line, PCR was simplified. Comparing with PrimeSTAR polymerase and Ultra-Super competent cell kit substitution, Quikchange kit protocol was more economical. Results： Objective recombinants were successfully gained by all three methods, so recombinant vectors could be utilized in further test. Conclusions： With two key elements substitution, Quikchang site-directed mutagenesis protocol could be improved to an effective, convenient, simple and economic approach.
作者机构：
[Deng, Min; Yang, Shuai; He, Xiu-Sheng; Luo, Qiao; Hu, Bo] Univ S China, Canc Res Inst, Hengyang City 421001, Hunan, Peoples R China.;[He, Zhi-Min; Xiao, Zhi-Qiang; Chen, Zhu-Chu] Cent S Univ, Canc Res Inst, Changsha 410078, Hunan, Peoples R China.
通讯机构：
[He, Xiu-Sheng] U;Univ S China, Canc Res Inst, Hengyang City 421001, Hunan, Peoples R China.
关键词：
Nasopharyngeal neoplasm;STGC3;Gene expression;Tet-on system
摘要：
STGC3 is a novel candidate tumor suppressor gene that was found to be associated with nasopharyngeal carcinoma (NPC) via the cDNA cloning and RACE processes. The biological function of the STGC3 protein and its expression level in nasopharyngeal carcinoma remain unknown. This study aimed to evaluate the STGC3 protein expression level in NPC and to investigate the inhibitory function of STGC3 as a candidate tumor suppressor gene. We assessed the expression of the STGC3 protein in NPC biopsies and normal control specimens via Western blot and immunohistochemical analysis. The expression of STGC3 as induced by doxycycline (Dox) via a tetracycline (Tet)-regulated system in human nasopharyngeal carcinoma cell line CNE2 was also established, and the effect of STGC3 restoration on the biological behavior of CNE2 was observed. A reduced level of STGC3 expression (0.978 +/- 0.213 versus 0.324 +/- 0.185, P < 0.05) was detected in NPC versus normal nasopharyngeal tissue by Western blot assay. Immunohistochemical assays for STGC3 detected positive staining in the nuclei and cytoplasm of epithelial cells, and the positive expression rate in NPC, 8 of 21 (38%), was lower than that in normal nasopharynx samples, 16 of 22 (72%). After STGC3 expression was restored, the growth capacity and clone formation potential of CNE2 cells in soft agar were significantly suppressed, and the cell percentage in G(0)/G(1) phase increased, while the percentage of cells entering the S and G(2) phases decreased. This indicates that an abnormality in STGC3 expression is associated with nasopharyngeal carcinogenesis and that it may play an important role in controlling cell growth and regulating the cell cycle.
作者：
Hu Bo;Qiu Qing-Chao;He Xiu-Sheng*;Luo Qiao;Tang Guo-Hua;...
期刊：
生物化学与生物物理进展,2007年34(5):538-545 ISSN：1000-3282
通讯作者：
He Xiu-Sheng
作者机构：
[Liao Yin-Hua; Luo Qiao; Qiu Qing-Chao; He Xiu-Sheng; He, XS; Hu Bo; Long Zhi-Feng; Tang Guo-Hua] Nanhua Univ, Canc Res Inst, Hengyang 421001, Peoples R China.
通讯机构：
[He Xiu-Sheng] N;Nanhua Univ, Canc Res Inst, Hengyang 421001, Peoples R China.
