TO THE EDITOR Sir, I read with great interest the recently published article in the World Journal of Gastroenterology by Jin and co-workers on the cloning and characterization of porcine aquaporin 1 water channel from...TO THE EDITOR Sir, I read with great interest the recently published article in the World Journal of Gastroenterology by Jin and co-workers on the cloning and characterization of porcine aquaporin 1 water channel from the pig liver and studies on its expression in the porcine gastrointestinal system. The authors should be congratulated for making this important and valuable contribution to the field of aquaporin biology and porcine gastrointestinal physiology. However, there are a number of unresolved issues and controversies concerning the expression of aquaporins （especially aquaporin 1） in the gastrointestinal system that are worthy of additional comment and discussion by Jin and co-workers.展开更多
The authors investigated the regulation of human aquaporin I（hAQP1） and the involvement of aquaporin 1 （AQP 1） in the migration of human hepatocellular carcinoma SMMC-7221 cells using RNA intereference technology ...The authors investigated the regulation of human aquaporin I（hAQP1） and the involvement of aquaporin 1 （AQP 1） in the migration of human hepatocellular carcinoma SMMC-7221 cells using RNA intereference technology Firstly, two short hairpin RNA（shRNA） constructs in PBSU6 vector were reconstructed and their knockdown effects were identified in SMMC-7221 cells. Next, the involvement of endogenous hAQP1 in regulating the migration of SMMC-7221 cells was investigated via siRNA technology. HAQPI-shRNA can specifically inhibit AQP1 dependent osmotic water permeability. Meanwhile the migration of SMMC-7221 cells was inhibited remarkably after silencing AQP1 by performing transwell cell migration assay and in vitro wound healing assay. Furthermore, in the presence of an inhibitor HgCl2, the water permeability of the cell membrane was remarkably decreased, the expression of AQP1 was upregulated after HgCla treatment and the cell movement was decreased at the moment. Increased AQP1 cannot attenuate cell migration ability when cell membrane loses its water permeability function. This demonstrates that the cell migration was remarkably related to the transporting water function of cell membrane.展开更多
Aquaporins（AQPs） are specific membrane channels for water and other small nonionic molecules.In order to overcome the difficulties to generate the effictive antibody of membrane protein,we selected the cytoplasmic C...Aquaporins（AQPs） are specific membrane channels for water and other small nonionic molecules.In order to overcome the difficulties to generate the effictive antibody of membrane protein,we selected the cytoplasmic C-terminus of Aquaporin 1（AQP1） as an unique antigen.The long C-terminus of mouse AQP1 was overexpressed in the Glutathione S-tansferase Gene Fusion System.On the basis of the resonable amounts of soluable membrane protein peptides,we prepared the specific antibody.To pursure this object,we constructed pGEX-4T-1/mAQP1（DNA sequence from 700 to 801 bp） recombinant plasmid and transformed it into Escherichia coli BL21 cells.The GST-AQP1 C-terminal hydrophilic peptide fusion protein was induced by IPTG and further purified by Glutathione Sepharose 4B to obtain the right size fusion protein.Then we immunized the New Zealand rabbits to prepare the antiserum.The purified AQP1 antibody showed high sensitivity by ELISA assay and high specificity by Western blot with AQP1 null mice served as negative control.Finally,we also checked the AQP1 localization in the mouse renal tissues in wild type of mice and AQP1 null mice served as negative control.We demonstrated that AQP1 was highly expressed at the descending limb of Henle tube using our purified AQP1 antibody,which was consistent with previous report.The successful design and preparation of AQP1 antibody through GST technique is an example as making antibodies against a specific membrane protein.展开更多
Objective： To investigate the expression of aquaporin 1 in cervical squamous carcinomas （CSC） and cervical precancerous lesions, and the relationship between the tumor clinicopathological parameters, prognosis and ...Objective： To investigate the expression of aquaporin 1 in cervical squamous carcinomas （CSC） and cervical precancerous lesions, and the relationship between the tumor clinicopathological parameters, prognosis and the expression of AQP1. Methods： Immunohistochemical method （EliVision） was used to detect the expression of AQP1 in samples from 106 patients [20 with normal cervical tissue, 30 with cervical intraepithelial neoplasia （stage Ⅰ and Ⅱ） and 56 with CSC]. Survival analysis was performed by Kaplan-Meier method. Results： AQP1 protein was expressed in vascular endothelia of all samples. It showed upregulation of AQP1 expression in CSC. There was a significant difference between CSC and normal cervical tissues （P〈0.05）. AQP1 was expressed in some tumor cells and unexpressed in normal squamous epithelial cells. And APQl-expressing tumor cells were positively related to lymph node metastasis. Patients with APQl-expressing tumor cells had the lower survival rate than the ones without. Conclusion： Abnormal expression of AQP1 plays an important role in the development of CSC. Positive expression of AQP1 in tumor cells maybe enhances tumor metastasis and could be used as a marker for tumor prognosis.展开更多
文摘[目的]研究腹水草(Veronicastrum axillare,V.axillare)含药血清对乙醇损伤的人胃上皮细胞(gastric epithelial cells-1,GES-1)的保护作用及其对环腺苷酸依赖性激酶(protein kinase A,PKA)、环磷腺苷效应元件结合蛋白(c AMP-response element binding protein,CREB)、水通道蛋白1(aquaporin 1,AQP1)表达水平的调控作用。[方法]20只雄性SD大鼠随机分为5组(n=4),连续灌胃给药14d,制备正常组(生理盐水20 m L·kg-1)、雷尼替丁组(0.027g·kg-1)以及V.axillare低、中、高剂量组(0.7、1.4、2.8 g·kg-1)含药血清。MTT法检测体外培养的GES-1细胞的活性,研究V.axillare含药血清及PKA抑制剂H-89(25μmol·m L-1)预处理对5%乙醇诱导GES-1细胞活力的影响。荧光定量RT-PCR检测PKA、CREB及AQP1 m RNA的表达水平,Western blot检测AQP1蛋白表达水平。[结果]模型组和H-89组GES-1细胞活性低于正常组,差异有统计学意义(P<0.01);乙醇造模处理显著上调PKA、CREB及下调AQP1的表达水平,差异有统计学意义(P<0.01);H-89处理显著抑制PKA和CREB m RNA表达水平(P<0.01);加入各剂量V.axillare含药血清不能显著改善H-89对PKA、CREB和AQP1 m RNA表达水平的抑制作用。V.axillare中、高剂量含药血清细胞活性高于模型组,PKA、CREB m RNA表达水平低于模型组,差异有统计学意义(P<0.01),而V.axillare中、低剂量含药血清AQP1表达水平高于模型组,差异有统计学意义(P<0.01)。[结论]V.axillare含药血清显著减轻乙醇对GES-1细胞的损伤,对GES-1细胞具有保护作用,其保护机制与下调PKA、CREB及上调AQP1的表达有关。
文摘TO THE EDITOR Sir, I read with great interest the recently published article in the World Journal of Gastroenterology by Jin and co-workers on the cloning and characterization of porcine aquaporin 1 water channel from the pig liver and studies on its expression in the porcine gastrointestinal system. The authors should be congratulated for making this important and valuable contribution to the field of aquaporin biology and porcine gastrointestinal physiology. However, there are a number of unresolved issues and controversies concerning the expression of aquaporins （especially aquaporin 1） in the gastrointestinal system that are worthy of additional comment and discussion by Jin and co-workers.
基金Supported by the National Natural Science Foundation of China(Nos 30871301,30700827)the Program of Ministry of Science and Technology of China(No2010DFA31430)+1 种基金the Program of Jilin Provincial Science & Technology Department, China(Nos20070719, 20080731 and 200905116)the Fund of Northeast Normal University, China(NoNENU-STC07005)
文摘The authors investigated the regulation of human aquaporin I（hAQP1） and the involvement of aquaporin 1 （AQP 1） in the migration of human hepatocellular carcinoma SMMC-7221 cells using RNA intereference technology Firstly, two short hairpin RNA（shRNA） constructs in PBSU6 vector were reconstructed and their knockdown effects were identified in SMMC-7221 cells. Next, the involvement of endogenous hAQP1 in regulating the migration of SMMC-7221 cells was investigated via siRNA technology. HAQPI-shRNA can specifically inhibit AQP1 dependent osmotic water permeability. Meanwhile the migration of SMMC-7221 cells was inhibited remarkably after silencing AQP1 by performing transwell cell migration assay and in vitro wound healing assay. Furthermore, in the presence of an inhibitor HgCl2, the water permeability of the cell membrane was remarkably decreased, the expression of AQP1 was upregulated after HgCla treatment and the cell movement was decreased at the moment. Increased AQP1 cannot attenuate cell migration ability when cell membrane loses its water permeability function. This demonstrates that the cell migration was remarkably related to the transporting water function of cell membrane.
基金Supported by the National Natural Science Foundation of China(Nos.30700827 and 30871301)Jilin Provincial Science & Technology Department of China(Nos.20070719 and 20080731)Northeast Normal University,China(Nos.20070401,NENU-STC07005)
文摘Aquaporins（AQPs） are specific membrane channels for water and other small nonionic molecules.In order to overcome the difficulties to generate the effictive antibody of membrane protein,we selected the cytoplasmic C-terminus of Aquaporin 1（AQP1） as an unique antigen.The long C-terminus of mouse AQP1 was overexpressed in the Glutathione S-tansferase Gene Fusion System.On the basis of the resonable amounts of soluable membrane protein peptides,we prepared the specific antibody.To pursure this object,we constructed pGEX-4T-1/mAQP1（DNA sequence from 700 to 801 bp） recombinant plasmid and transformed it into Escherichia coli BL21 cells.The GST-AQP1 C-terminal hydrophilic peptide fusion protein was induced by IPTG and further purified by Glutathione Sepharose 4B to obtain the right size fusion protein.Then we immunized the New Zealand rabbits to prepare the antiserum.The purified AQP1 antibody showed high sensitivity by ELISA assay and high specificity by Western blot with AQP1 null mice served as negative control.Finally,we also checked the AQP1 localization in the mouse renal tissues in wild type of mice and AQP1 null mice served as negative control.We demonstrated that AQP1 was highly expressed at the descending limb of Henle tube using our purified AQP1 antibody,which was consistent with previous report.The successful design and preparation of AQP1 antibody through GST technique is an example as making antibodies against a specific membrane protein.
基金the Fund of Science and Technology Development Program of Shaanxi Province (2005K09-G6, 2005K09-G9)
文摘Objective： To investigate the expression of aquaporin 1 in cervical squamous carcinomas （CSC） and cervical precancerous lesions, and the relationship between the tumor clinicopathological parameters, prognosis and the expression of AQP1. Methods： Immunohistochemical method （EliVision） was used to detect the expression of AQP1 in samples from 106 patients [20 with normal cervical tissue, 30 with cervical intraepithelial neoplasia （stage Ⅰ and Ⅱ） and 56 with CSC]. Survival analysis was performed by Kaplan-Meier method. Results： AQP1 protein was expressed in vascular endothelia of all samples. It showed upregulation of AQP1 expression in CSC. There was a significant difference between CSC and normal cervical tissues （P〈0.05）. AQP1 was expressed in some tumor cells and unexpressed in normal squamous epithelial cells. And APQl-expressing tumor cells were positively related to lymph node metastasis. Patients with APQl-expressing tumor cells had the lower survival rate than the ones without. Conclusion： Abnormal expression of AQP1 plays an important role in the development of CSC. Positive expression of AQP1 in tumor cells maybe enhances tumor metastasis and could be used as a marker for tumor prognosis.
