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Qing-Wei Li,
Xiao-Yan Lu,
Yong You, Huan Sun,
Xin-Yu Liu,
Jian-Zhong Ai,
Rui-Zhi Tan,
Tie-Lin Chen,
Mian-Zhi Chen,
Hong-Lian Wang,
Yu-Quan Wei,
Qin Zhou
[show abstract]
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ABSTRACT: Autosomal recessive polycystic kidney disease (ARPKD), characterized by ectatic collecting duct, is an infantile form of PKD occurring in 1 in 20 000 births. Despite having been studied for many years, little is known about the underlying mechanisms. In the current study, we employed, for the first time, a MS-based comparative proteomics approach to investigate the differently expressed proteins between kidney tissue samples of four ARPKD and five control individuals. Thirty two differently expressed proteins were identified and six of the identified protein encoding genes performed on an independent group (three ARPKD subjects, four control subjects) were verified by semi-quantitative RT-PCR, and part of them were further validated by Western blot and immunohistochemistry. Moreover, similar alteration tendency was detected after downregulation of PKHD1 by small interfering RNA in HEK293T cell. Interestingly, most of the identified proteins are associated with mitochondria. This implies that mitochondria may be implicated in ARPKD. Furthermore, the String software was utilized to investigate the biological association network, which is based on known and predicted protein interactions. In conclusion, our findings depicted a global understanding of ARPKD progression and provided a promising resource of targeting protein, and shed some light further investigation of ARPKD.
Proteomics 06/2012; 12(15-16):2556-70. · 4.43 Impact Factor
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Jingjing Duan,
Huizhe Huang,
Xiaoyan Lv,
Honglian Wang,
Ziwei Tang, Huan Sun,
Qingwei Li,
Jianzhong Ai,
Ruizhi Tan,
Yuhang Liu,
Mianzhi Chen,
Weiwei Duan,
Yuquan Wei,
Qin Zhou
[show abstract]
[hide abstract]
ABSTRACT: Autosomal recessive polycystic kidney disease (ARPKD) is a severe inherited disorder with an incidence of 1/20 000 live births. Mutations of PKHD1 (polycystic kidney and hepatic disease gene 1) gene were identified to be responsible for ARPKD. However, the underlying molecular mechanisms remain largely unknown. MicroRNAs (miRNAs) are an abundant class of small RNAs with global effect on gene expression. Up to 30% of human protein coding genes may be regulated by miRNAs. However, to date, nothing is known regarding the role of miRNAs in PKHD1. In this study, we exploited bioinformatics to analyse the 3'UTR of PKHD1 gene and illustrated that the 3'UTR region of the gene is highly conserved in evolution. We identified about 35 candidate miRNAs within a 3738 bp window of the 3'UTR region. Of the 35 potential miRNAs, miR-365-1 emerged to post-transcriptionally modulate the expression of PKHD1. Furthermore, we demonstrated that miR-365-1 modulated PKHD1 suppressed cell-cell adhesion in part through E-cadherin.
Cell Biochemistry and Function 03/2012; 30(5):382-9. · 1.77 Impact Factor
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ABSTRACT: MicroRNAs (miRNAs) have gradually been recognized as regulators of embryonic development; however, relatively few miRNAs have been identified that regulate cardiac development. A series of recent papers have established an essential role for the miRNA-17-92 (miR-17-92) cluster of miRNAs in the development of the heart. Previous research has shown that the Friend of Gata-2 (FOG-2) is critical for cardiac development. To investigate the possibility that the miR-17-92 cluster regulates FOG-2 expression and inhibits proliferation in mouse embryonic cardiomyocytes we initially used bioinformatics to analyze 3' untranslated regions (3'UTR) of FOG-2 to predict the potential of miR-17-92 to target it. We used luciferase assays to demonstrate that miR-17-5p and miR-20a of miR-17-92 interact with the predicted target sites in the 3'UTR of FOG-2. Furthermore, RT-PCR and Western blot were used to demonstrate the post-transcriptional regulation of FOG-2 by miR-17-92 in embryonic cardiomyocytes from E12.5-day pregnant C57BL/6J mice. Finally, EdU cell assays together with the FOG-2 rescue strategy were employed to evaluate the effect of proliferation on embryonic cardiomyocytes. We first found that the miR-17-5p and miR-20a of miR-17-92 directly target the 3'UTR of FOG-2 and post-transcriptionally repress the expression of FOG-2. Moreover, our findings demonstrated that over-expression of miR-17-92 may inhibit cell proliferation via post-transcriptional repression of FOG-2 in embryonic cardiomyocytes. These results indicate that the miR-17-92 cluster regulates the expression of FOG-2 protein and suggest that the miR-17-92 cluster might play an important role in heart development.
Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas / Sociedade Brasileira de Biofisica ... [et al.] 02/2012; 45(2):131-8. · 1.08 Impact Factor
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Jianzhong Ai,
Huizhe Huang,
Xiaoyan Lv,
Ziwei Tang,
Mianzhi Chen,
Tielin Chen,
Weiwei Duan, Huan Sun,
Qingwei Li,
Ruizhi Tan,
Yuhang Liu,
Jingjing Duan,
Yang Yang,
Yuquan Wei,
Yan Li,
Qin Zhou
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ABSTRACT: Hepatocellular carcinoma (HCC) is one of the most deadly diseases; metastasis and recurrence are the most important factors that affect the therapy of HCC chronically. Until now, the prognosis for the metastasis of HCC had not improved. Recently, several proteins that are related to metastasis and invasion of HCC were identified, but the effective markers still remain to be elucidated.
In this study, comparative proteomics was used to study the differentially expressed proteins in two HCC cell lines MHCC97L and HCCLM9, which have low and high metastatic potentials, respectively.
Our findings indicated that filamin A (FLNA) and phosphoglycerate kinase 1 (PGK1) were two significantly differentially expressed proteins, with high expression in HCCLM9 cells, and may influence the metastasis of HCC cells.
Taken together with the confirmation of expression on the mRNA level, we propose the use of FLNA and PGK1 as potential markers for the progression of HCC.
Cellular Physiology and Biochemistry 01/2011; 27(3-4):207-16. · 2.86 Impact Factor
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Yan Xiao,
Xiaoyan Lv,
Ge Cao,
Guohui Bian,
Jingjing Duan,
Jianzhong Ai, Huan Sun,
Qingwei Li,
Qiutan Yang,
Tielin Chen,
Danhua Zhao,
Ruizhi Tan,
Yuhang Liu,
Yidong Wang,
Zheng Zhang,
Yang Yang,
Yuquan Wei,
Qin Zhou
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ABSTRACT: Trpp5 is one member of the polycystic kidney disease (PKD) family, which belongs to transient receptor potential (TRP) superfamily. Our previous study has shown that Trpp5 is developmentally expressed in mouse testis and overexpression of Trpp5 increases intracellular free calcium concentration in MDCK cells. However, the roles of this protein in cellular processes are largely unknown. Here, we demonstrated that Trpp5 resided in both cytoplasm and cell membrane of HEK293 cells. We found that overexpression of Trpp5 slightly increased the calcium current amplitude of HEK293 cells and shifted the reversal potential to a more negative value. Meanwhile, overexpression of Trpp5 suppressed proliferation of Hela cells via inhibiting DNA replication and induced apoptosis of Hela cells with morphological changes and accumulation of fragmented DNA. Collectively, these findings suggest that Trpp5 might involve calcium homeostasis contributing to cell proliferation and apoptosis.
Molecular and Cellular Biochemistry 06/2010; 339(1-2):155-61. · 2.06 Impact Factor
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Jianzhong Ai,
Jingjing Duan,
Xiaoyan Lv,
Mianzhi Chen,
Qiutan Yang, Huan Sun,
Qingwei Li,
Yan Xiao,
Yidong Wang,
Zheng Zhang,
Ruizhi Tan,
Yuhang Liu,
Danhua Zhao,
Tielin Chen,
Yang Yang,
Yuquan Wei,
Qin Zhou
[show abstract]
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ABSTRACT: Multiple lines of evidence have shown that the functional defect of pancreatic β cells is the root cause of type 2 diabetes. FoxO1, a key transcription factor of fundamental cellular physiology and functions, has been implicated in this process. However, the underlying molecular mechanism is still largely unknown. Here, we show that the overexpression of FoxO1 promotes the proliferation of cultured pancreatic β cells exposed to low nutrition, while no change in apoptosis was observed compared with the control group. Moreover, by using two specific inhibitors for PI3K and MAPK signaling, we found that FoxO1 might be the downstream transcription factor of these two pathways. Furthermore, a luciferase assay demonstrated that FoxO1 could regulate the expression of Ccnd1 at the transcription level. Collectively, our findings indicated that FoxO1 modulated by both MAPK and PI3K signaling pathways was prone to cause the proliferation, but not the apoptosis, of pancreatic β cells exposed to low nutrition, at least partially, by regulating the expression of Ccnd1 at the transcription level.
12/2009;
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Jianzhong Ai,
Jingjing Duan,
Xiaoyan Lv,
Mianzhi Chen,
Qiutan Yang, Huan Sun,
Qingwei Li,
Yan Xiao,
Yidong Wang,
Zheng Zhang,
Ruizhi Tan,
Yuhang Liu,
Danhua Zhao,
Tielin Chen,
Yang Yang,
Yuquan Wei,
Qin Zhou
[show abstract]
[hide abstract]
ABSTRACT: Multiple lines of evidence have shown that the functional defect of pancreatic beta cells is the root cause of type 2 diabetes. FoxO1, a key transcription factor of fundamental cellular physiology and functions, has been implicated in this process. However, the underlying molecular mechanism is still largely unknown. Here, we show that the overexpression of FoxO1 promotes the proliferation of cultured pancreatic beta cells exposed to low nutrition, while no change in apoptosis was observed compared with the control group. Moreover, by using two specific inhibitors for PI3K and MAPK signaling, we found that FoxO1 might be the downstream transcription factor of these two pathways. Furthermore, a luciferase assay demonstrated that FoxO1 could regulate the expression of Ccnd1 at the transcription level. Collectively, our findings indicated that FoxO1 modulated by both MAPK and PI3K signaling pathways was prone to cause the proliferation, but not the apoptosis, of pancreatic beta cells exposed to low nutrition, at least partially, by regulating the expression of Ccnd1 at the transcription level.
Biochemistry 11/2009; 49(1):218-25. · 3.42 Impact Factor
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Huan Sun,
Qing-Wei Li,
Xiao-Yan Lv,
Jian-Zhong Ai,
Qiu-Tan Yang,
Jing-Jing Duan,
Guo-Hui Bian,
Yan Xiao,
Yi-Dong Wang,
Zheng Zhang,
Yu-Hang Liu,
Rui-Zhi Tan,
Yang Yang,
Yu-Quan Wei,
Qin Zhou
[show abstract]
[hide abstract]
ABSTRACT: To identify the possible microRNAs (miRNAs) which target the polycystic kidney disease-2 gene (PKD2), and clarify effects of the miRNAs on PKD2. We preliminarily used bioinformatics to analyze 3'UTR (3'untranslated regions) of PKD1 and PKD2 in order to predict the potential microRNAs targeted on them. Subsequently, the stable cell lines with overexpression of microRNA-17 (miR-17) were screened, and luciferase assay combined with the mutation 3'UTR of PKD2 were performed to verify PKD2 is the target of miR-17. Moreover, RT-PCR and Western Blotting were used to determine the post-transcriptionally regulation of PKD2 by miR-17. Finally, MTT cell assays allied with PKD2 rescued strategy were employed to evaluate cell proliferation effects. Our study firstly found that the 3'UTR of PKD2 was more conservation than that of PKD1, and microRNA-17 directly targets the 3'UTR of PKD2 and post-transcriptionally repress the expression of PKD2. Moreover, our findings also demonstrated that overexpression of miR-17 may promote cell proliferation via post-transcriptionally repression of PKD2 in HEK 293T. This suggested that microRNA might be a novel mechanism for cystogenesis as well as a potential therapeutic target for the cell proliferation of autosomal dominant polycystic kidney disease (ADPKD).
Molecular Biology Reports 10/2009; 37(6):2951-8. · 2.93 Impact Factor
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Guo-Hui Bian,
Ge Cao,
Xiao-Yan Lv,
Qing-Wei Li, Huan Sun,
Yan Xiao,
Jian-Zhong Ai,
Qui-Tan Yang,
Jing-Jing Duan,
Yi-Dong Wang,
Zheng Zhang,
Rui-Zhi Tan,
Yu-Hang Liu,
Yang Yang,
Yu-Quan Wei,
Qin Zhou
[show abstract]
[hide abstract]
ABSTRACT: The Pkd2 gene encodes an integral protein (~130 kDa), named polycystin-2 (PC-2). PC-2 is mainly involved in autosomal dominant polycystic kidney disease. Recently, polycystin-1/polycystin-2 complex has been shown to act as an adhesion complex mediating or regulating cell-cell or cell-matrix adhesion, suggesting that PC-2 may play a role in cell-cell/cell-matrix interactions. Here, we knocked down the expression of Pkd2 gene with small interfering RNAs (siRNAs) in the mouse melanoma cells (B16 cells), indicating that the cells transfected with the targeted siRNAs significantly suppressed cell-cell adhesion, but not cell-matrix adhesion, compared to the cells transfected with non-targeted control (NC) siRNA. This study provides the first directly functional evidence that PC-2 mediates cell-cell adhesion. Furthermore, we demonstrated that PC-2 modulated cell-cell adhesion may be, at least partially, associated with E-cadherin. Collectively, these findings for the first time showed that PC-2 may mediate cell-cell adhesion, at least partially, through E-cadherin.
Molecular Biology Reports 09/2009; 37(5):2387-95. · 2.93 Impact Factor
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ABSTRACT: PKD2 gene encodes a critical cation channel protein that plays important roles in various developmental processes and is usually evolutionarily conserved. In the present study, we analyzed the evolutionary patterns of PKD2 and its homologous genes (PKD2L1, PKD2L2) from nine mammalian species. In this study, we demonstrated the orthologs of PKD2 gene family evolved under a dominant purifying selection force. Our results in combination with the reported evidences from functional researches suggested the entire PKD2 gene family are conserved and perform essential biological roles during mammalian evolution. In rodents, PKD2 gene family members appeared to have evolved more rapidly than other mammalian lineages, probably resulting from relaxation of purifying selection. However, positive selection imposed on synonymous sites also potentially contributed to this case. For the paralogs, our results implied that PKD2L2 genes evolved under a weaker purifying selection constraint than PKD2 and PKD2L1 genes. Interestingly, some loop regions of transmembrane domain of PKD2L2 exhibited higher P (N)/P (S) ratios than expected, suggesting these regions are more functional divergent in organisms and worthy of special attention.
Genetica 03/2009; 137(1):77-86. · 2.15 Impact Factor
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Zheng Zhang,
Xiaoyan Lv,
Yidong Wang,
Ye Chen,
Rong Zheng, Huan Sun,
Guohui Bian,
Yan Xiao,
Qingwei Li,
Qiutan Yang,
Jianzhong Ai,
Jingjing Duan,
Ruizhi Tan,
Yuhang Liu,
Yang Yang,
Yuquan Wei,
Qin Zhou
[show abstract]
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ABSTRACT: Embryos transfer is one of the most critical steps for generating genetically modified mice. Because of the limitations of the current transfer equipment and techniques, such as discharge of freshly transferred embryos and the necessity of sophisticated transfer skills, the success ratio of implantation may not well satisfy the requirements for mass production of high quality animals in the field of genetically modified mice. In this study, we describe a modified embryo transfer pipette with a syringe-like tip that can easily be applied to mouse embryo transfer. Using this improved method, we show that the procedures for mouse embryo implantation are simplified and easier to perform; moreover, the viability rate of mouse embryos is 20% higher than that achieved with conventional methods. Our modified tool and improved transfer technique are effective, time-saving and less invasive, resulting in increased success of embryo transfer.
Journal of Reproduction and Development 12/2008; 55(1):94-7. · 1.46 Impact Factor
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Yi-Dong Wang,
Guo-Hui Bian,
Xiao-Yan Lv,
Rong Zheng, Huan Sun,
Zheng Zhang,
Ye Chen,
Qin-Wei Li,
Yan Xiao,
Qiu-Tan Yang,
Jian-Zhong Ai,
Yu-Quan Wei,
Qin Zhou
[show abstract]
[hide abstract]
ABSTRACT: Although previous studies have implicated a role for TC1 (C8orf4) in cancer cell proliferation, the molecular mechanism of its action is still largely unclear. In this study, we showed, for the first time, that the mRNA levels of TC1 were upregulated by mitogens (FBS/thrombin) and at least partially, through the ERK1/2 signaling pathway. Interestingly, the over-expression of TC1 promoted the G(1)- to S-phase transition of the cell cycle, which was delayed by the deficiency of ERK1/2 signaling in fibroblast cells. Furthermore, the luciferase reporter assay indicated that the over-expression of TC1 significantly increased Cyclin D1 promoter-driven luciferase activity. Taken together, our findings revealed that TC1 was involved in the mitogen-activated ERK1/2 signaling pathway and positively regulated G(1)- to S-phase transition of the cell cycle. Our results may provide a novel mechanism of the role of TC1 in the regulation of cell proliferation.
BMB reports 11/2008; 41(10):733-8. · 1.72 Impact Factor
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Ye Chen,
Zheng Zhang,
Xiao-Yan Lv,
Yi-Dong Wang,
Zhong-Guo Hu, Huan Sun,
Rui-Zhi Tan,
Yu-Hang Liu,
Guo-Hui Bian,
Yan Xiao,
Qin-Wei Li,
Qiu-Tan Yang,
Jian-Zhong Ai,
Lu Feng,
Yang Yang,
Yu-Quan Wei,
Qin Zhou
[show abstract]
[hide abstract]
ABSTRACT: Pkd2l2 is a novel member of the polycystic kidney disease (PKD) gene family in mammals. Prominently expressed in testis, this gene is still poorly understood. In this study, reverse transcription polymerase chain reaction (RT-PCR) results showed a time-dependent expression pattern of Pkd2l2 in postnatal mouse testis. Immunohistochemical analysis revealed that Pkd2l2 encoded a protein, polycystin-L2, which was predominantly detectable in the plasma membrane of spermatocytes and round spermatids, as well as in the head and tail of elongating spermatids within seminiferous tubules in mouse testis tissue sections of postnatal day 14 and adult mice. A green fluorescent fusion protein of Pkd2l2 resided in the plasma membrane of HEK 293 and MDCK cells, suggesting that it functions as a plasma membrane protein. Overexpression of Pkd2l2 increased the intracellular calcium concentration of MDCK cells, as detected by flow cytometry. Collectively, these data indicated that Pkd2l2 may be involved in the mid-late stage of spermatogenesis through modulation of the intracellular calcium concentration.
Biological & Pharmaceutical Bulletin 09/2008; 31(8):1496-500. · 1.66 Impact Factor
