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
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
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
The correlation between aquaporin 1 （AQP1） and hypoxia-inducible factor 1 （HIF 1） in breast cancer tissues was preliminarily studied. In 155 cases of breast cancer, the expression levels of AQP1 were detected by i...The correlation between aquaporin 1 （AQP1） and hypoxia-inducible factor 1 （HIF 1） in breast cancer tissues was preliminarily studied. In 155 cases of breast cancer, the expression levels of AQP1 were detected by immunohistochemisty in HIFl-positive group or HIFl-negative group, and the correlation between AQP1 and HIF1 was analyzed. The overexpression of AQP1 and HIF1 were observed in 155 cases of breast cancer tissues. The expression level of AQP1 in HIFl-positive group was significantly higher than that in HIFl-negative group. The positive expression rate of AQP1 was 296.55±24.67 and 168.37±37.53 in HIFl-positive group and HIFl-negative group respectively with the difference being very significant between them （P〈0.001）. It was concluded that AQP1 was overexpressed in the HIFl-positive group and there were some correlations between AQP1 and HIF1, suggesting they interact each other and regulate the oncogenesis of breast cancer.展开更多
Background The glioblastoma has served as a valuable experimental model system for investigating the growth and invasive properties of glioblastoma.Aquaporin-1(AQP1)in facilitating cell migration and potentially contr...Background The glioblastoma has served as a valuable experimental model system for investigating the growth and invasive properties of glioblastoma.Aquaporin-1(AQP1)in facilitating cell migration and potentially contributing to tumor progression.In this study,we analyzed the role of AQP1 overexpression in glioblastoma and elucidated the main mechanisms involved.Methods AQP1 overexpression recombinant vector was introduced into C6 rat glioma cells to construct an AQP1 overexpression C6 cell line,and its effect on cell viability and migration ability was detected by MTT and Transwell.RNA was extracted by Trizol method for gene sequencing and transcriptomics analysis,and the differentially expressed genes(DEGs)were enriched for up-and downregulated genes by Principal component analysis(PCA),and the molecular mechanism of AQP1 overexpression was analyzed in comparison with the control group using the NCBI GEO database.Statistical analysis was performed using Mann-Whitney paired two tailed t test.Results The cell viability of AQP1-transfected cell lines increased by 23%and the mean distance traveled increased by 67%compared with the control group.Quantitative analysis of gene expression showed that there were 12,121 genes with an average transcripts per million(TPM)value greater than 1.DEGs accounted for 13%of the genes expressed,with the highest correlation with upregulated genes being FOXO4 and MAZ,and the highest with down-regulated genes being E2F TFs.Conclusions AQP1 may be implicated in glioma formation by interacting with the transcriptional regulation networks involving the FOXO4,MAZ,and E2F1/2.These findings shed light on the potential significance of AQP1 in glioma pathogenesis and warrant further investigations to unravel the underlying molecular mechanisms.展开更多
Background Aquaporin-1 （AQP1） has involved in fluid transport in diverse pulmonary edema diseases. Our study aimed to explore the dynamic changes of AQP1 in pulmonary water metabolism in rats following traumatic bra...Background Aquaporin-1 （AQP1） has involved in fluid transport in diverse pulmonary edema diseases. Our study aimed to explore the dynamic changes of AQP1 in pulmonary water metabolism in rats following traumatic brain injury （TBI） and the protective effect provided by shenmai injection.Methods Sixty male Sprague Dawley rats weighting 280-300 g were randomly divided into three groups： the normal control group, the model group and the shenrnai injection （SMI） group. One piece skull was taken away without injuring cerebral tissue in normal control group, while rats in model group and SMI group were subject to free fall injury in the cerebral hemisphere. Rats in model group received intraperitoneal normal sodium （15 mi/kg） at one hour post-injury and the same dose of shenmai injection instead in SMI group, respectively. The expression of AQP1 was detected by immunohistochemical analysis and semi-quantitative RT-PCR at 0 hour, 10 hours, 72 hours and 120 hours after TBI.Arterial blood gas analysis and lung wet to dry were also measured.Results AQP1 was mainly presented in the capillary endothelium and slightly alveolar epithelial cells in three groups, but the expression of AQP1 in the normal control group was positive and tenuous, weakly positive in the model and SMI groups,respectively. Compared with normal control group, AQP1 mRNA levels were down regulated in the model and SMI groups at 10 hours, 72 hours and 120 hours （P ￡1/40.05）. While AQP1 mRNA levels in the SMI group was up-regulated than that in the model group （P￡1/40.05）. Lung wet to dry weight ratio （W/D） in the model and SMI groups at 10 hours were higher than that in normal control group （P ￡1/40.05）. Compared with normal control group, PaO2 was markedly lower in the model and SMI groups （P ￡1/40.05）, but there were no statistically significant differences between model and SMI groups （P ￡3/40.05）.Conclusions The decreased AQP1 expression may be involved in the increased lung water content and dysfunction of pulmonary water metabolism following TBI. The treatment with SMI could improve water metabolism by promoting AQP1 expression.展开更多
The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were stu...The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were studied, and the implication of AQP1 regulation in corticosteroid-glaucoma and the possibility of AS-ODN inhibiting the AQP1 expression were evaluated. The cultured HTM cells in vitro were treated with different concentrations of dexamethasone and transfected with oligonucleotides for 5 days respectively. Then, total RNA and protein of HTM cells were extracted. The changes of AQP1 mRNA and protein were demonstrated qualitatively and quantitatively by RT-PCR and Western blot. Band intensities were detected by imaging analysis. There was a parallel relationship between the results of RT-PCR and those of Western blot. The expression levels of AQP1 mRNA and protein in dexamethasone-treated groups were increased initially and decreased later as dexamethasone concentration was stepped up. In the 0.04 μg/mL and 0.4 μg/mL groups, the levels of AQP1 were higher than in control group （0 μg/mL）. In the 4 μg/ mL and 40 μg/mL groups, the AQP1 expression levels were lower than in control group. AS-ODN could down-regulate the expression of AQP1 mRNA and protein in a dose-dependent manner. At 5 μg/mL, down-regulation efficiency reached the maximum. There was no statistically significant difference in the expression of AQP1 mRNA and protein between all sense oligonucleotides groups and control group. It was suggested that dexamethasone may induce the changes of the AQP1 expression in HTM cells to be involved in the occurrence of corticosteroid-glaucoma. AS-ODN can down-regulate the AQP1 expression in HTM cells to some extent.展开更多
AIM：To investigate the role of Aquaporin-1（AQP-1）in lens epithelial cells（LECs）and its potential target genes.AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparen...AIM：To investigate the role of Aquaporin-1（AQP-1）in lens epithelial cells（LECs）and its potential target genes.AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparency maintenance.Herein,AQP-1 expression in LECs was investigated to evaluate its influence on cell survival in association with its potential role in cataract formation.·M ETHODS：LECs were transfected with lentivirus carrying AQP-1 small interfering RNA（si RNA）.Real-time polymerase chain reaction（PCR）and Western blotting were conducted to detect AQP-1 expression in LECs from different groups.Meanwhile,cell counting kit-8（CCK-8）assay and flow cytometry were performed to measure LEC proliferation and apoptosis,respectively.·RESULTS：AQP-1 expression was significantly reduced in LECs,both at m RNA and protein levels（〈0.05）,after si RNA treatment.Decreased cell viability was detected by CCK-8 assay in LECs with si RNA interference,compared to control cells（〈0.05）.The apoptosis rate significantly increased in cells after si RNA interference（〈0.05）.·CONCLUSION：The decreased cell viability following AQP-1 down regulation is largely due to its induction of apoptosis of LECs.AQP-1 reduction might lead to changes of physiological functions in LECs,which might be associated with the occurrence and development of cataracts.展开更多
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.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.
文摘The correlation between aquaporin 1 （AQP1） and hypoxia-inducible factor 1 （HIF 1） in breast cancer tissues was preliminarily studied. In 155 cases of breast cancer, the expression levels of AQP1 were detected by immunohistochemisty in HIFl-positive group or HIFl-negative group, and the correlation between AQP1 and HIF1 was analyzed. The overexpression of AQP1 and HIF1 were observed in 155 cases of breast cancer tissues. The expression level of AQP1 in HIFl-positive group was significantly higher than that in HIFl-negative group. The positive expression rate of AQP1 was 296.55±24.67 and 168.37±37.53 in HIFl-positive group and HIFl-negative group respectively with the difference being very significant between them （P〈0.001）. It was concluded that AQP1 was overexpressed in the HIFl-positive group and there were some correlations between AQP1 and HIF1, suggesting they interact each other and regulate the oncogenesis of breast cancer.
基金supported by Natural Science Foundation of Hainan Province(821RC692)Hainan Provincial Health Industry Research Project(20A200136)Hainan Natural Science Foundation(2019RC388)
文摘Background The glioblastoma has served as a valuable experimental model system for investigating the growth and invasive properties of glioblastoma.Aquaporin-1(AQP1)in facilitating cell migration and potentially contributing to tumor progression.In this study,we analyzed the role of AQP1 overexpression in glioblastoma and elucidated the main mechanisms involved.Methods AQP1 overexpression recombinant vector was introduced into C6 rat glioma cells to construct an AQP1 overexpression C6 cell line,and its effect on cell viability and migration ability was detected by MTT and Transwell.RNA was extracted by Trizol method for gene sequencing and transcriptomics analysis,and the differentially expressed genes(DEGs)were enriched for up-and downregulated genes by Principal component analysis(PCA),and the molecular mechanism of AQP1 overexpression was analyzed in comparison with the control group using the NCBI GEO database.Statistical analysis was performed using Mann-Whitney paired two tailed t test.Results The cell viability of AQP1-transfected cell lines increased by 23%and the mean distance traveled increased by 67%compared with the control group.Quantitative analysis of gene expression showed that there were 12,121 genes with an average transcripts per million(TPM)value greater than 1.DEGs accounted for 13%of the genes expressed,with the highest correlation with upregulated genes being FOXO4 and MAZ,and the highest with down-regulated genes being E2F TFs.Conclusions AQP1 may be implicated in glioma formation by interacting with the transcriptional regulation networks involving the FOXO4,MAZ,and E2F1/2.These findings shed light on the potential significance of AQP1 in glioma pathogenesis and warrant further investigations to unravel the underlying molecular mechanisms.
文摘Background Aquaporin-1 （AQP1） has involved in fluid transport in diverse pulmonary edema diseases. Our study aimed to explore the dynamic changes of AQP1 in pulmonary water metabolism in rats following traumatic brain injury （TBI） and the protective effect provided by shenmai injection.Methods Sixty male Sprague Dawley rats weighting 280-300 g were randomly divided into three groups： the normal control group, the model group and the shenrnai injection （SMI） group. One piece skull was taken away without injuring cerebral tissue in normal control group, while rats in model group and SMI group were subject to free fall injury in the cerebral hemisphere. Rats in model group received intraperitoneal normal sodium （15 mi/kg） at one hour post-injury and the same dose of shenmai injection instead in SMI group, respectively. The expression of AQP1 was detected by immunohistochemical analysis and semi-quantitative RT-PCR at 0 hour, 10 hours, 72 hours and 120 hours after TBI.Arterial blood gas analysis and lung wet to dry were also measured.Results AQP1 was mainly presented in the capillary endothelium and slightly alveolar epithelial cells in three groups, but the expression of AQP1 in the normal control group was positive and tenuous, weakly positive in the model and SMI groups,respectively. Compared with normal control group, AQP1 mRNA levels were down regulated in the model and SMI groups at 10 hours, 72 hours and 120 hours （P ￡1/40.05）. While AQP1 mRNA levels in the SMI group was up-regulated than that in the model group （P￡1/40.05）. Lung wet to dry weight ratio （W/D） in the model and SMI groups at 10 hours were higher than that in normal control group （P ￡1/40.05）. Compared with normal control group, PaO2 was markedly lower in the model and SMI groups （P ￡1/40.05）, but there were no statistically significant differences between model and SMI groups （P ￡3/40.05）.Conclusions The decreased AQP1 expression may be involved in the increased lung water content and dysfunction of pulmonary water metabolism following TBI. The treatment with SMI could improve water metabolism by promoting AQP1 expression.
文摘The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were studied, and the implication of AQP1 regulation in corticosteroid-glaucoma and the possibility of AS-ODN inhibiting the AQP1 expression were evaluated. The cultured HTM cells in vitro were treated with different concentrations of dexamethasone and transfected with oligonucleotides for 5 days respectively. Then, total RNA and protein of HTM cells were extracted. The changes of AQP1 mRNA and protein were demonstrated qualitatively and quantitatively by RT-PCR and Western blot. Band intensities were detected by imaging analysis. There was a parallel relationship between the results of RT-PCR and those of Western blot. The expression levels of AQP1 mRNA and protein in dexamethasone-treated groups were increased initially and decreased later as dexamethasone concentration was stepped up. In the 0.04 μg/mL and 0.4 μg/mL groups, the levels of AQP1 were higher than in control group （0 μg/mL）. In the 4 μg/ mL and 40 μg/mL groups, the AQP1 expression levels were lower than in control group. AS-ODN could down-regulate the expression of AQP1 mRNA and protein in a dose-dependent manner. At 5 μg/mL, down-regulation efficiency reached the maximum. There was no statistically significant difference in the expression of AQP1 mRNA and protein between all sense oligonucleotides groups and control group. It was suggested that dexamethasone may induce the changes of the AQP1 expression in HTM cells to be involved in the occurrence of corticosteroid-glaucoma. AS-ODN can down-regulate the AQP1 expression in HTM cells to some extent.
基金Supported by the National Natural Science Foundation of China(No.81070715)Innovative Platform Foundation of Fujian ProvinceChina(No.2010Y2003)
文摘AIM：To investigate the role of Aquaporin-1（AQP-1）in lens epithelial cells（LECs）and its potential target genes.AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparency maintenance.Herein,AQP-1 expression in LECs was investigated to evaluate its influence on cell survival in association with its potential role in cataract formation.·M ETHODS：LECs were transfected with lentivirus carrying AQP-1 small interfering RNA（si RNA）.Real-time polymerase chain reaction（PCR）and Western blotting were conducted to detect AQP-1 expression in LECs from different groups.Meanwhile,cell counting kit-8（CCK-8）assay and flow cytometry were performed to measure LEC proliferation and apoptosis,respectively.·RESULTS：AQP-1 expression was significantly reduced in LECs,both at m RNA and protein levels（〈0.05）,after si RNA treatment.Decreased cell viability was detected by CCK-8 assay in LECs with si RNA interference,compared to control cells（〈0.05）.The apoptosis rate significantly increased in cells after si RNA interference（〈0.05）.·CONCLUSION：The decreased cell viability following AQP-1 down regulation is largely due to its induction of apoptosis of LECs.AQP-1 reduction might lead to changes of physiological functions in LECs,which might be associated with the occurrence and development of cataracts.
