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ABSTRACT: miR-21 is the most commonly over-expressed microRNA (miRNA) in cancer and a proven oncogene. Hsa-miR-21 is located on chromosome 17q23.2, immediately downstream of the vacuole membrane protein-1 (VMP1) gene, also known as TMEM49. VMP1 transcripts initiate ∼ 130 kb upstream of miR-21, are spliced, and polyadenylated only a few hundred base pairs upstream of the miR-21 hairpin. On the other hand, primary miR-21 transcripts (pri-miR-21) originate within the last introns of VMP1, but bypass VMP1 polyadenylation signals to include the miR-21 hairpin. Here, we report that VMP1 transcripts can also bypass these polyadenylation signals to include miR-21, thus providing a novel and independently regulated source of miR-21, termed VMP1-miR-21. Northern blotting, gene-specific RT-PCR, RNA pull-down and DNA branching assays support that VMP1-miR-21 is expressed at significant levels in a number of cancer cell lines and that it is processed by the Microprocessor complex to produce mature miR-21. VMP1 and pri-miR-21 are induced by common stimuli, such as phorbol-12-myristate-13-acetate (PMA) and androgens, but show differential responses to some stimuli such as epigenetic modifying agents. Collectively, these results indicate that miR-21 is a unique miRNA capable of being regulated by alternative polyadenylation and two independent gene promoters.
Nucleic Acids Research 04/2012; 40(14):6821-33. · 8.03 Impact Factor
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ABSTRACT: Dose-escalated radiation therapy for localized prostate cancer (PCa) has a clear therapeutic benefit; however, escalated doses may also increase injury to noncancerous tissues. Radiosensitizing agents can improve ionizing radiation (IR) potency, but without targeted delivery, these agents will also sensitize surrounding normal tissues. Here we describe the development of prostate-targeted RNAi agents that selectively sensitized prostate-specific membrane antigen-positive (PSMA-positive) cells to IR. siRNA library screens identified DNA-activated protein kinase, catalytic polypeptide (DNAPK) as an ideal radiosensitization target. DNAPK shRNAs, delivered by PSMA-targeting RNA aptamers, selectively reduced DNAPK in PCa cells, xenografts, and human prostate tissues. Aptamer-targeted DNAPK shRNAs, combined with IR, dramatically and specifically enhanced PSMA-positive tumor response to IR. These findings support aptamer-shRNA chimeras as selective sensitizing agents for the improved treatment of high-risk localized PCa.
The Journal of clinical investigation 06/2011; 121(6):2383-90. · 15.39 Impact Factor
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Juxiang Chen,
Qihan Wu,
Yicheng Lu,
Tao Xu,
Yan Huang,
Judit Ribas, Xiaohua Ni,
Guohan Hu,
Fengping Huang,
Liangfu Zhou,
Daru Lu
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ABSTRACT: SPP1 was found to be significantly upregulated in many kinds of malignant tumors, including gliomas. Considering that gene polymorphisms have been implicated in the development of gliomas, we performed an association study between SPP1 functional promoter region polymorphisms and glioma risk in a Chinese population. We found significant evidence of an association between SPP1 promoter polymorphisms and glioma risk. For the -155_156insG variant, the -155_156GG allele was found to be significantly associated with an increased risk of glioma (P=0.020, odds ratio (OR)=1.202, 95% confidence interval (CI): 1.028-1.408). Individuals with the genotype containing the GG allele had a 1.372-fold increased risk (P=0.006, OR=1.372, 95% CI: 1.095-1.719). Further stratified analyses suggested that a significant association existed in adult glioma patients, male subjects and in cases without a family history of cancer. Alternatively, the study of single-nucleotide polymorphism -443C/T in a recessive model revealed that the genotype CC+CT significantly decreased the risk of glioma when compared with TT (P=0.023, OR=0.774, 95% CI: 0.621-0.966). After the analysis of haplotypes, the haplotype -155_156GG/-443T was represented at a significantly higher frequency in cases (P=0.029, OR=1.192, 95% CI: 1.018-1.395). Cellular assay indicated that the transcriptional activity of the SPP1 promoter containing the -155_156GG allele significantly increased in glioma cells. Thus, variants of the SPP1 promoter might influence the risk of glioma by regulating promoter activity. Further analyses are necessary to validate our observation in larger samples or in other ethnic groups.
Journal of Human Genetics 07/2010; 55(7):456-61. · 2.57 Impact Factor
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ABSTRACT: MicroRNAs (miRNAs) are small non-coding RNAs that play important roles in regulation of eukaryotic gene expression. Aberrant expression and structural alternation of miRNAs are considered to participate in tumorigenesis and cancer development. Recently, different genotypes of miR-196a polymorphisms (SNP, rs11614913) were found to be associated with the survival of patients with lung cancer and increased risk of breast cancer. To further investigate whether this polymorphism may influence glioma risk or not, we examined the SNP allele frequency in Chinese population. Our data shows the genotype CC of miR-196a (rs11614913) polymorphism is associated with decreased risk of glioma in the Chinese population (OR = 0.74, 95% CI:0.56-0.98). Furthermore, a significant association was observed between this genotype and glioma risk in the subgroups of adult glioma (OR = 0.73, 95% CI:0.55-0.98), male glioma (OR = 0.69, 95% CI:0.48-0.99) and patients with glioblastoma (OR = 0.58, 95% CI:0.37-0.91). This was the first study investigating the association between the miR-196a rs11614913 and glioma risk. Compared with the results from previous studies in lung cancer and breast cancer, our data suggest a different genotype association in glioma. This may be related to the diversity on the tissue origin, tumor type, tumorigenesis, and developing process.
Journal of Cancer Research and Clinical Oncology 03/2010; 136(12):1853-9. · 2.56 Impact Factor
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Judit Ribas, Xiaohua Ni,
Michael Haffner,
Erik A Wentzel,
Amirali Hassanzadeh Salmasi,
Wasim H Chowdhury,
Tarana A Kudrolli,
Srinivasan Yegnasubramanian,
Jun Luo,
Ron Rodriguez,
Joshua T Mendell,
Shawn E Lupold
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ABSTRACT: Androgen receptor (AR)-mediated oncogenic pathways have not been fully elucidated. In this study, we used high-throughput microarray analysis on two AR-positive prostate cancer (CaP) cell lines to identify 16 AR-responsive microRNAs (miRNA). We focused on miR-21 because of its previously reported oncogenic activity in other cancers. We show androgen-induced AR binding to the defined miR-21 promoter, miPPR-21, suggesting direct transcriptional regulation. Inhibition of miR-21 diminished androgen-induced CaP cell proliferation, providing new evidence that miRNAs can contribute to androgen-driven cell growth. Elevated expression of miR-21 enhanced CaP tumor growth in vivo and, surprisingly, was sufficient for androgen-dependent tumors to overcome castration-mediated growth arrest. Thus, elevated miR-21 expression alone is sufficient to impart castration resistance. Moreover, quantitative reverse transcription-PCR analysis revealed elevated miR-21 expression in CaP when compared with adjacent normal tissue. These results suggest that miR-21 may contribute to CaP pathogenesis.
Cancer Research 10/2009; 69(18):7165-9. · 7.86 Impact Factor
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Juxiang Chen,
Gang Yin,
Yicheng Lu,
Meiqing Lou,
Haipeng Cheng, Xiaohua Ni,
Guohan Hu,
Chun Luo,
Kang Ying,
Yi Xie,
Yumin Mao
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ABSTRACT: J-domain proteins are involved in protein importing and sorting, protein translocation into the endoplasmic reticulum and interaction with hsp70, cell cycle regulation and exocytosis. During the large-scale sequencing analysis of a human fetal brain cDNA library, we isolated a cDNA clone encoding a putative 531 amino acid protein with a DnaJ domain and C2H2-type zinc finger motif, which was named as DNAJA5 (human DnaJ homology subfamily A member 5). The putative protein of DNAJA5 gene was homologous to Caenorhabditis elegans DNJ-17 (identity 33.2% similarity 45.1%). By mapping, we located DNAJA5 gene to human chromosome 5p12-5p13. The DNAJA5 gene contained at least 12 exons that spanned more than 25.6 kb of the genomic sequence. The reverse transcription-polymerase chain reaction analysis indicated that it was expressed in the human brain, placenta, kidney and pancreas. DNAJA5 might play a role in co-operating with Hsp70.
International Journal of Molecular Medicine 06/2004; 13(5):735-40. · 1.98 Impact Factor
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Gang Yin,
JianLiang Dai,
Chaoneng Ji, Xiaohua Ni,
Guang Shu,
Xin Ye,
Jianfeng Dai,
Qihan Wu,
Shaohua Gu,
Yi Xie,
Robert Chunhua Zhao,
Yumin Mao
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ABSTRACT: Intraflagellar transport (IFT), in which molecular motors and IFT particle proteins participate, is very important in assembling and maintaining many cilia/flagella, such as the motile cilia that drive the swimming of cells and embryos, the nodal cilia that generate left-right asymmetry in vertebrate embryos, and the sensory cilia that detect sensory stimuli in some animals. Here we report the molecular cloning and characterization of a novel human cDNA that is 1060 base pair in length, encoding a putative protein of 158 amino acid residues which shares high homology to Mus musculus intraflagellar transport protein mIFT20 (Accession number AAL99202, identity 82.3%, similarity 82.9%) at protein level. Bioinformatics analysis revealed that the gene is composed of 6 exons and is located on human chromosome 17p 11.1. RT-PCR showed that this gene is expressed in human brain, lung, kidney and pancreas, and lower expression were also detected in human placenta, liver, thymus, prostate and testis.
Molecular Biology Reports 01/2004; 30(4):255-60. · 2.93 Impact Factor
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Haipeng Cheng,
Qi Gao,
Min Jiang,
Yushu Ma, Xiaohua Ni,
Lingchen Guo,
Wei Jin,
Gentao Cao,
Chaoneng Ji,
Kang Ying,
Weiwen Xu,
Shaohua Gu,
Yuhong Ma,
Yi Xie,
Yumin Mao
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ABSTRACT: Reversible phosphorylation is recognized to be a major mechanism for the control of intracellular events in eukaryotic cells. From a human fetal brain cDNA library, we isolated a cDNA clone encoding a novel dual specificity protein phosphatase, which showed 88% identity with previously reported mouse LMW-DSP3 at the amino acid level. The deduced protein had a single dual-specificity phosphatase catalytic domain, and lacked a cdc25 homology domain. LMW-DSP3 was expressed in the heart, lung, liver, and pancreas, and the expression level in the pancreas was highest. The LMW-DSP3 gene was located in human chromosome 2q32, and consisted of five exons spanning 21kb of human genomic DNA. LMW-DSP3 fused to GST showed phosphatase activity towards p-nitrophenyl phosphate which was optimal at pH 7.0 and 40 degrees C, and the activity was enhanced by Ca(2+) and Mn(2+). The phosphatase activity of LMW-DSP3 was inhibited by orthovanate. LMW-DSP3 showed phosphatase activity toward oligopeptides containing pSer/Thr and pTyr, indicating that LMW-DSP3 is a protein phosphatase with dual substrate specificity.
The International Journal of Biochemistry & Cell Biology 03/2003; 35(2):226-34. · 4.63 Impact Factor
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Juxiang Chen,
Yan Huang,
Hai Wu, Xiaohua Ni,
Haipeng Cheng,
Jingping Fan,
Shaohua Gu,
Xing Gu,
Gentao Cao,
Kang Ying,
Yumin Mao,
Yicheng Lu,
Yi Xie
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ABSTRACT: The J-domain is believed to be part of a chaperone involved in protein folding. From a fetal brain cDNA library, we isolated a cDNA of 3249 bp encoding a novel human J-domain protein, which was named as HDJ3. The expression pattern of HDJ3 was examined by reverse transcription/polymerase chain reaction, which suggested that the transcripts were highly expressed in human pancreas and selectively expressed in human brain, lung, liver, skeletal muscle and kidney. The results also showed that a probable splice variant of HDJ3 gene might exist. The HDJ3 gene was located on human chromosome 12q13.1-12q13.2 and consisted of seven exons spanning 8593 bp of the human genome. PSORT analysis indicated that the HDJ3 gene contained a transmembrane domain. The putative protein of the HDJ3 gene was highly homologous to rat dopamine-receptor-interacting protein, suggesting that it was a novel member of the molecular chaperone family and functionally related to dopamine signal transduction.
Journal of Human Genetics 02/2003; 48(5):217-21. · 2.57 Impact Factor
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ABSTRACT: The NM23 gene is a conspicuous metastasis-suppressor gene. Eight human genes of the NM23/nucleoside diphosphate kinase family have been discovered. From our large cDNA cloning and sequencing project, we cloned a different transcript ( NM23-H1B) of human NM23-H1 from 18-week-old human fetal brain. The 987-bp cDNA encodes a protein of 177 amino acid residues. Compared with NM23H1, the cDNA contained an additional NH(2)-terminal region (25 amino acid residues). It was mapped to chromosome 17q21.3 using bioinformatics analysis, which shows that the second exon does not exist in NM23-H1. The expression pattern of NM23-H1B showed that it was ubiquitously expressed in normal tissues (15 tissues except colon) at different levels. Our data also indicated that the expression of the transcript in tumors related to tumor differentiation: in poorly differentiated breast carcinoma GI-101, pancreatic adenocarcinoma GI-103, and undifferentiated ovarian carcinoma GI-102, there was no expression. In poorly differentiated lung carcinoma LX-1, lung carcinoma GI-117, the expression level was very low. The transcript band in well-differentiated colon adenocarcinoma CX-1 was significantly higher than that in poorly differentiated colon adenocarcinoma GI-112. A high transcription level was also found in grade IV prostatic adenocarcinoma PC3.
Journal of Human Genetics 02/2003; 48(2):96-100. · 2.57 Impact Factor
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ABSTRACT: Protein 4.1 is an important structural protein that is expressed in erythroid and in a variety of nonerythroid tissues. In mammalian erythrocytes, it plays a key role in regulating the physical properties of mechanical stability and deformability in membranes by stabilizing the spectrin-actin interaction. The protein 4.1 family mainly comprises 4.1R, 4.1G (general type), 4.1B (brain type), and 4.1N (neuron type). We identified a novel human 4.1 ( 4.1O) gene that is 2312 bp in length and encodes a protein of 553 amino acid residues. The protein shared homology with mouse protein 4.1B (identity 38%, similarity 55%) with a FERM domain. The expression pattern of the human 4.1O gene in 16 tissues showed that there was a transcript only in ovary, whereas in the remaining 15 tissues, specific bands of the transcript could not be detected. In eight human fetal tissues, the specific bands of the transcript could be detected in skeletal muscle, with lower levels detected in thymus and brain. The 4.1O gene consists of 14 exons and 13 introns and was mapped to Chromosome 9q21-9q22 by bioinformatics analysis.
Journal of Human Genetics 02/2003; 48(2):101-6. · 2.57 Impact Factor
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ABSTRACT: Rab proteins are small molecular weight GTPases that control vesicular traffic in eukaryotic cells. A subset of Rab proteins, the Rab3 proteins are thought to play an important role in regulated exocytosis of vesicles. During the large-scale sequencing analysis of a human fetal brain cDNA library, we isolated a cDNA clone encoding a novel Rab protein, which showed 99% identity with previously isolated bovine Rab3C at the amino acid level It contained four conserved motifs characteristic of the Rab3 family. RT-PCR analysis indicated that human Rab3C was expressed in the human brain, placenta, and lung. By mapping, we localized the Rab3Cgene to human chromosome 5q13. The Rab3C gene consisted of 6 exons spanning more than 310 kb of human genomic DNA. Rab3A, Rab3B, and Rab3D have been mapped to three different chromosomes, suggesting that they are not transcripts of the same gene.
Biochemical Genetics 09/2002; 40(7-8):263-72. · 0.86 Impact Factor
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Haipeng Cheng,
Yushu Ma, Xiaohua Ni,
Min Jiang,
Lingchen Guo,
Wei Jin,
Weiwen Xu,
Gentao Cao,
Chaoneng Ji,
Kang Yin,
Shaohua Gu,
Yuhong Ma,
Yi Xie,
Yumin Mao
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ABSTRACT: F-box proteins are an expanding family of eukaryotic proteins that are characterized by an approximately 40 amino acid motif. Some F-box proteins are critical for the controlled degradation of cellular regulatory proteins. During a large-scale sequencing analysis of a human fetal brain cDNA library, we isolated a cDNA clone that encodes a novel F-box protein. It showed a 90.0% identity with the previously isolated mouse F-box protein16 at the amino acid level. Northern blot analysis showed no detectable expression, while reverse transcription-polymerase chain reaction analysis indicated that FBXO16 was expressed in the heart, spleen, and colon. By mapping, we localized the FBXO16 gene to the human chromosome 8p12. The FBXO16 gene consisted of 9 exons that spanned 67,816 bp of human genomic DNA.
Molecules and Cells 09/2002; 14(1):56-9. · 2.18 Impact Factor
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ABSTRACT: We isolated and sequenced a cDNA clone coding a human protein kinase CK1 (casein kinase 1) by screening a human fetal brain cDNA library. This new cDNA clone of 1756 bp contained an open reading frame, encoding a protein of 438 amino acids with a molecular weight of 50,272 Da and an isoelectric point of 9.37. The entire amino acid sequence of the novel human CK1 was 94% homologous to that of rat CK1gamma1. Northern blot analysis indicated that the human CK1gamma1 was highly expressed in the liver, skeletal muscle, heart and kidney. Furthermore, the human CK1gamma1 gene was mapped to chromosome 15q22 between STS marker D15S159 and D15S125 by polymerase chain reaction analysis of human/rodent hybrid cell panels.
International Journal of Molecular Medicine 09/2002; 10(2):227-30. · 1.98 Impact Factor
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ABSTRACT: We report cloning and some features of a novel human gene, MATH2, which encodes a protein of 337 amino acid residues with a basic helix loop helix domain and exhibits 98% similarity to mouse Math2. Results of Northern blot analysis revealed two transcripts of the MATH2 gene of 1.7 kb and 2.4 kb in human brain. We localized MATH2 to chromosome 7 at 7p14-15 by matching with the Human Genome Sequence Database. Human MATH2 and mouse Math2 may have the same functions in the nervous system.
Journal of Genetics 05/2002; 81(1):13-7. · 1.09 Impact Factor
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ABSTRACT: Pantothenate kinase (PanK) is a key regulatory enzyme in the CoA biosynthetic pathway in bacteria and mammalian cells. It catalyzes the first committed step in the universal biosynthetic pathway leading to CoA. Here we report the molecular cloning and characterization of a new human PanK gene. The gene encoded a protein of 314 amino acid residues, which share high homology to mouse pantothenate kinase (mPanK) 1 beta. Northern blot analysis revealed a transcript of the gene of 2.6 kb in human liver and kidney. The human PanK gene was located to human chromosome between 10q22.3 and 10q23.2 by bioinformatics analysis.
The International Journal of Biochemistry & Cell Biology 03/2002; 34(2):109-15. · 4.63 Impact Factor
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ABSTRACT: Rab proteins are small-molecular-weight guanosine triphosphatases (GTPases) that control vesicular traffic in eukaryotic cells. The small GTPase Rab2 is a resident of pre-Golgi intermediates and is required for protein transport from the endoplasmic reticulum to the Golgi complex. We identified a novel human Rab (Rab2B) gene that was 2312 bp in length and encoded a protein of 216 amino acid residues. The protein shared high homology with mouse Rab2 (identity 83%, similarity 91%). The expression pattern of the human Rab2B gene showed that there is a transcript in kidney, prostate, lung, liver, thymus, colon, pancreas, and skeletal muscle, and low levels in placenta, whereas specific bands of the transcript could not be detected in heart, brain, spleen, testis, ovary, small intestine, and leukocyte. Overexpression has been observed in colon adenocarcinoma CX-1. The Rab2B gene consists of nine exons and eight introns and is mapped to chromosome 14q11.1-14q11.2 by bioinformatics analysis.
Journal of Human Genetics 02/2002; 47(10):548-51. · 2.57 Impact Factor
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ABSTRACT: Cyclic adenosine monophosphate (cAMP) response element-binding (CREB) proteins are a family of mammalian transcription activators. We identified a novel human CREB gene ( CREB4) that was 1592 bp long and encoded a protein of 395 amino acid residues. The protein shared high homology to mouse CREB3 (identity 62%, similarity 72%). The expression pattern of the human CREB4 gene showed transcripts in prostate, brain, pancreas, skeletal muscle, small intestine, testis, leukocyte, and thymus, whereas in heart, lung, liver, kidney, placenta, spleen, ovary, and colon, specific bands of the transcript could not be detected. The CREB4 gene consisted of ten exons and nine introns and was mapped to chromosome 1q21.3 by means of a bioinformatics analysis.
Journal of Human Genetics 02/2002; 47(7):373-6. · 2.57 Impact Factor
