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Qianjin Liao,
Xiaofang Guo,
Xiaoling Li,
Wei Xiong,
Xiayu Li,
Jing Yang,
Pan Chen,
Wenling Zhang,
Haibo Yu,
Hailin Tang,
Min Deng,
Fang Liang,
Minghua Wu,
Zhaohui Luo, Rong Wang,
Xi Zeng,
Zhaoyang Zeng,
Guiyuan Li
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ABSTRACT: Prohibitin-1 (PHB, also known as PHB1) is a pleiotropic protein in cells. PHB is a cell-surface receptor and is involved in the regulation of proliferation, apoptosis, transcription, and mitochondrial protein folding. PHB is upregulated in 5-8F cells, which overexpress LPLUNC1 (long palate, lung, nasal epithelium clone 1, a candidate tumour suppressor gene), and was identified using two-dimensional fluorescence difference gel electrophoresis and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS/MS). Thus, we examined PHB mRNA levels using 24 nasopharyngeal carcinoma (NPC) and eight normal nasopharyngeal epithelium (NPE) tissues. Protein levels were detected using immunohistochemistry with a tissue microarray consisting of 323 NPC and NPE tissues. A Kaplan-Meier analysis was carried out, and the log-rank test was used to determine the statistical significance of the results using SPSS 15.0 software. PHB mRNA and protein expression levels were significantly downregulated in NPC tissue specimens compared with the NPE samples (P<0.01). In addition, decreased PHB expression correlates significantly with a poor prognosis, whereas decreased PHB protein expression is closely associated with advanced clinical stage and metastasis in NPC lesions. Therefore, we favour the hypothesis that the expression level of PHB could be used as a potential prognostic biomarker for NPC.
European journal of cancer prevention: the official journal of the European Cancer Prevention Organisation (ECP) 06/2012; · 2.21 Impact Factor
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Zuping Zhang,
Hailin Tang,
Zeyou Wang,
Baoxin Zhang,
Wei Liu,
Hongmei Lu,
Lan Xiao,
Xiaoping Liu, Rong Wang,
Xiaoling Li,
Minghua Wu,
Guiyuan Li
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ABSTRACT: Perturbation of DNA methylation is frequent in cancers and has emerged as an important mechanism involved in tumorigenesis. To determine how DNA methylation is modified in the genome of primary glioma, we used Methyl-DNA immunoprecipitation (MeDIP) and Nimblegen CpG promoter microarrays to identify differentially DNA methylation sequences between primary glioma and normal brain tissue samples.
MeDIP-chip technology was used to investigate the whole-genome differential methylation patterns in glioma and normal brain tissues. Subsequently, the promoter methylation status of eight candidate genes was validated in 40 glioma samples and 4 cell lines by Sequenom's MassARRAY system. Then, the epigenetically regulated expression of these genes and the potential mechanisms were examined by chromatin immunoprecipitation and quantitative real-time PCR.
A total of 524 hypermethylated and 104 hypomethylated regions were identified in glioma. Among them, 216 hypermethylated and 60 hypomethylated regions were mapped to the promoters of known genes related to a variety of important cellular processes. Eight promoter-hypermethylated genes (ANKDD1A, GAD1, HIST1H3E, PCDHA8, PCDHA13, PHOX2B, SIX3, and SST) were confirmed in primary glioma and cell lines. Aberrant promoter methylation and changed histone modifications were associated with their reduced expression in glioma. In addition, we found loss of heterozygosity (LOH) at the miR-185 locus located in the 22q11.2 in glioma and induction of miR-185 over-expression reduced global DNA methylation and induced the expression of the promoter-hypermethylated genes in glioma cells by directly targeting the DNA methyltransferases 1.
These comprehensive data may provide new insights into the epigenetic pathogenesis of human gliomas.
Molecular Cancer 09/2011; 10:124. · 3.99 Impact Factor
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Xiaofang Guo,
Qianjin Liao,
Pan Chen,
Xiayu Li,
Wei Xiong,
Jian Ma,
Xiaoling Li,
Zhaohui Luo,
Hailin Tang,
Min Deng,
Yin Zheng, Rong Wang,
Wenling Zhang,
Guiyuan Li
[show abstract]
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ABSTRACT: Dysregulation of microRNA (miRNA) metabolism has been observed in a variety of human cancers, but the expression patterns of the enzymes responsible for generating miRNAs remain largely unexplored. In this study, we investigated the expression profiles of the two most important enzymes of the miRNA machinery, Drosha and Dicer, which were closely correlated with nasopharyngeal carcinoma (NPC) and patient survival.
Dicer and Drosha mRNA levels were detected by quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) using 24 NPC tissues, 7 normal nasopharyngeal epithelium samples (NPE) and NPC cell lines. In addition, protein levels were detected by immunohistochemistry (IHC) using an NPC tissue microarray (TMA), which include 251 NPC and 105 NPE cases. For some NPC patients can not be contacted, the survival data were available only for 146 patients. Kaplan-Meier analysis was performed, and the chi-square and log-rank tests were used to detect significance levels using SPSS 15.0 software.
The mean level of Dicer and Drosha mRNA were significantly down-regulated in NPC tissue specimens and cell lines when compared with controls. The low levels of Dicer and Drosha protein were frequently seen in NPC, and the low expression of Dicer and Drosha protein was significantly correlated with shorter progression-free survival (PFS) and overall survival (OS) of NPC patients.
We observed that Drosha and Dicer expression was dysregulation in NPC compared with healthy control samples and was significantly correlated with shorter PFS and OS of NPC patients. Therefore, we hypothesise that the expression levels of Dicer and Drosha could be used as potential prognostic biomarkers for NPC.
Journal of Cancer Research and Clinical Oncology 09/2011; 138(1):49-56. · 2.56 Impact Factor
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ABSTRACT: LRRC4 is a tumor suppressor of glioma, and it is epigenetically inactivated commonly in glioma. Our previous study has shown that induction of LRRC4 expression inhibits the proliferation of glioma cells. However, little is known about the mechanisms underlying the action of LRRC4 in glioma cells. We employed two-dimensional fluorescence differential gel electrophoresis (2-D DIGE) and MALDI -TOF/TOF-MS/MS to identify 11 differentially expressed proteins, including the significantly down-regulated STMN1 expression in the LRRC4-expressing U251 glioma cells. The levels of STMN1 expression appeared to be positively associated with the pathogenic degrees of human glioma. Furthermore, induction of LRRC4 over-expression inhibited the STMN1 expression and U251 cell proliferation in vitro, and the glioma growth in vivo. In addition, induction of LRRC4 or knockdown of STMN1 expression induced cell cycle arrest in U251 cells, which was associated with modulating the p21, cyclin D1, and cyclin B expression, and the ERK phosphorylation, and inhibiting the CDK5 and cdc2 kinase activities, but increasing the microtubulin polymerization in U251 cells. LRRC4, at least partially by down-regulating the STMN1expression, acts as a major glioma suppressor, induces cell cycle arrest and modulates the dynamic process of microtubulin, leading to the inhibition of glioma cell proliferation and growth. Potentially, modulation of LRRC4 or STMN1 expression may be useful for design of new therapies for the intervention of glioma.
Journal of Cellular Biochemistry 08/2011; 112(12):3621-9. · 2.87 Impact Factor
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Hailin Tang,
Xiaoping Liu,
Zeyou Wang,
Xiaoling She,
Xi Zeng,
Min Deng,
Qianjin Liao,
Xiaofang Guo, Rong Wang,
Xiaoling Li,
Fang Zeng,
Minghua Wu,
Guiyuan Li
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ABSTRACT: LRRC4 is not only a brain-specific gene, but it has also been identified as a tumor suppressor gene for glioma. Promoter methylation of LRRC4 is frequently involved in the inactivation in glioma. MiRNA-mediated gene regulation has recently been demonstrated to play an important role in multiple biological processes related to cancer, including glioma. In this study, we demonstrated that a small regulatory microRNA, hsa-miR-381, an "oncomir", had a major role in glioma progression and that LRRC4 was a target of hsa-miR-381. By regulating LRRC4, hsa-miR-381 increased the in vitro and in vivo proliferation of glioma cells, and this action was associated with decreased inhibition of MEK/ERK and AKT signaling. Conversely, LRRC4, as a glioma suppressor, inhibited the endogenous expression of hsa-miR-381 and decreased cell proliferation and tumor growth. The interaction of hsa-miR-381 and LRRC4 is involved in the pathogenesis of glioma. In addition, the stable expression of hsa-miR-381 in blood provides a novel and promising diagnostic biomarker, and anti-hsa-miR-381 "antagomir" may be an ideal target for glioma therapy.
Brain research 03/2011; 1390:21-32. · 2.46 Impact Factor
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Bo Xiang,
Mei Yi,
Li Wang,
Wei Liu,
Wenling Zhang,
Jue Ouyang,
Ya Peng,
Wenjuan Li,
Dachuan Yin,
Ming Zhou,
Huaying Liu,
Minghua Wu, Rong Wang,
Xiaoling Li,
Guiyuan Li
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ABSTRACT: Oxidored-nitro domain containing protein 1 (NOR1) gene is a novel nitroreductase gene first isolated from nasopharyngeal carcinoma (NPC). It plays an important role in the formation of chemical carcinogen and the carcinogenesis of NPC for its nitrosation function. Overexpression of the wild-type NOR1 gene in nasopharyngeal carcinoma cells is effective to inhibit cell growth and proliferation. In this study, for the first time, we generated a highly specific NOR1 antibody and analyzed NOR1 distribution in the human tissues and NPC biopsies. The results showed that NOR1 protein is predominantly expressed in human nasopharynx and tracheal tissues. Human heart, liver, spleen, stomach, colon, kidney, skeletal muscle, thymus, and pancreas are all deficient of NOR1 protein. More importantly, we performed immunohistochemistry assay of NOR1 protein expression in the NPC tissues, and the result showed that NOR1 protein is frequently down-expressed in NPC. These data shed light on the selectivity of potential physiological functions of NOR1 and provides an indispensable reference to the carcinogenesis process of NPC and to identify or validate tissue-specific drug targets.
Acta Biochimica et Biophysica Sinica 10/2009; 41(9):754-62. · 1.38 Impact Factor
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Wenling Zhang,
Zhaoyang Zeng,
Yanhong Zhou,
Wei Xiong,
Songqing Fan,
Lan Xiao,
Donghai Huang,
Zheng Li,
Dan Li,
Minghua Wu,
Xiaoling Li,
Shourong Shen, Rong Wang,
Li Cao,
Ke Tang,
Guiyuan Li
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ABSTRACT: Previous studies have revealed that Epstein-Barr virus (EBV) was closely associated with nasopharyngeal carcinoma (NPC). This study aimed to characterize the global pathways affected in the EBV-associated NPC. Combined with microdissection, gene expression profiles in 22 NPCs and 10 non-tumor nasopharyngeal epithelial (NPE) tissue samples were analyzed. All NPC specimens served in the microarray analysis were positive for EBV, as judged by identification of the expression of EBV nuclear antigen 1 (EBNA1). Through gene set enrichment analysis (GSEA), we found that cell cycle pathway was the most disregulated pathway in NPC (P=0.000, false discovery rate q-value=0.007), which included some aberrant expressed components. We first found that overexpression of CDK4, cyclin D1, and Rb proteins, and loss of expression of proteins p16, p27, and p19 were statistically significant in NPC tissues compared with non-cancerous NPE (P<0.05) by real-time RT-PCR and tissue microarray. EBV-encoded small RNA-1 (EBER-1) hybridization signals in the NPC showed significant associations with the overexpression of Rb (P=0.000), cyclin D1 (P=0.000), CDK4 (P=0.000), and the loss of expression of p16 proteins (P=0.039). In the final logistic regression analysis model, EBER-1 and abnormal expression of p16, Rb, cyclin D1, and E2F6 were independent contributions to nasopharyngeal carcinogenesis. Through survival analysis, only cyclin D1 could predict the prognosis of NPC patients. These results suggested that cell cycle pathway was the most disregulated pathway in the EBV-associated NPC, and EBER-1 was closely associated with p16, CDK4, cyclin D1, and Rb.cyclin D1 could be the prognosis biomarker for NPC.
Acta Biochimica et Biophysica Sinica 05/2009; 41(5):414-28. · 1.38 Impact Factor
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Ke Tang,
Li Cao,
Song-qing Fan,
Ming-hua Wu,
He Huang,
Yan-hong Zhou,
Ming Zhou,
Yun-lian Tang, Rong Wang,
Fang Zeng,
Ping Liao,
Xiao-ling Li,
Gui-yuan Li
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ABSTRACT: To explore the effect of all-trans-retinoic acid (ATRA) on the growth inhibition and cellular differentiation of C6 glioma cells.
Human glioma C6 cells were treated with 5 mg/L ATRA,and the inhibition of cell growth was assessed by methyl thiazolyl tetrazolium assay. The differentiation of C6 cells was determined by flow cytometry, microscopy,transmission electron microscope, and immunohistochemical technique.
Treatment of ATRA could result in the growth inhibition of C6 cells, and the cell density significantly decreased(P<0.01). The cell cycle distribution was changed, G0/G1 phase was prolonged, and cells at S phase decreased(P<0.01). The C6 glioma cells displayed normal fibroblast-like morphology under the microscope before the induction, and the ATRA-treated C6 cells became slightly long, turned into round in the middle, and had protrusions at both ends. The ATRA-treated C6 cells did not display obvious apoptosis by flow cytometry(P>0.05).Whereas, early apoptosis was observed under the transmission electron microscope, the vacuoles increased, the mitochondria and endoplasmic reticulum were abundant in the cytoplasm, and the cellular structures tended to be normal.The expression of glial fibrillaryacidic protein in C6 cells increased in the treatment group.
ATRA can inhibit the proliferation, and induce the differentiation of C6 glioma cells.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 10/2008; 33(10):892-7.
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Bingyi Xiao,
Songqing Fan,
Zhaoyang Zeng,
Wei Xiong,
Li Cao,
Yixin Yang,
Weifang Li, Rong Wang,
Ke Tang,
Jun Qian,
Shourong Shen,
Xiaoling Li,
Guiyuan Li
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ABSTRACT: Recently, the UBAPl gene, a putative nasopharyngeal carcinoma (NPC) related gene, which is located at human chromosome 9p21-22 where loss of heterozygosity frequently occurs in NPC, was cloned. The present study aimed to explore the purification approach to UBAP1 protein and its expression pattern in NPC with tissue microarray. The full length coding sequence of UBAP1 was subcloned into a prokaryotic expression vector, pGEX-4T-2, and expressed in Escherichia coli as a GST-fusion protein. With modification of the purification method, GST-UBAP1 fusion protein achieved a high level of purity. The New Zealand rabbit was immunized with the purified fusion protein to prepare polyclonal antiserum following standard protocols. With this antiserum, high-throughput analysis of UBAP1 protein expression using immunohistochemistry was performed on self-made tissue microarrays consisting of 316 nasopharyngeal specimens from 148 NPC and 168 non-cancerous nasopharyngeal epithelia with different morphological features. Consequently, we found that there was a significant decrease in the percentage of positive expression in all NPC on protein levels for UBAP1 (23%), compared to that of the non-NPC (89%) (P<0.01). The result suggests that UBAP1 might be a potential effective diagnosis candidate for NPC and decreased expression of UBAP1 protein is a possible point of dysfunction along the pathogenesis pathway for NPC that may contribute to malignant transformation.
Protein Expression and Purification 06/2006; 47(1):60-7. · 1.59 Impact Factor
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Wei Xiong,
Zhao Yang Zeng,
Jia Hui Xia,
Kun Xia,
Shou Rong Shen,
Xiao Ling Li,
Dong Xu Hu,
Chen Tan,
Juan Juan Xiang,
Jie Zhou, [......],
Yan Hong Zhou,
Xiao Min Luo,
Hou De Zhou,
Yi Xin Yang,
He Ping Dai,
Guo Yin Feng,
Qian Pan,
Ling Qian Wu,
Lin He,
Gui Yuan Li
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ABSTRACT: Nasopharyngeal carcinoma (NPC) poses one of the serious health problems in southern Chinese, with an incidence rate ranging from 15 to 50/100,000. Chromosome translocation t(1;3) and frequent loss of heterogeneity on short arms of chromosome 3 and 9 have been reported to be associated with NPC, and a genome-wide scan identified an NPC susceptibility locus on chromosome 4p15.1-q12 recently. In our study, we collected samples from 18 families at high risk of NPC from the Hunan province in southern China, genotyped with a panel of polymorphic markers on short arms of chromosomes 3, 9, and 4p15.1-q12. A locus on 3p21 was identified to link to NPC with a maximum logarithm of odds for linkage score of 4.18. Fine mapping located the locus to a 13.6-cM region on 3p21.31-21.2, where a tumor suppressor gene cluster resided. Our findings identified a novel locus for NPC and provided a map location for susceptibility genes candidates. In contrast to a recent study, no significant evidence for NPC linkage to chromosomes 4 and 9 was observed.
Cancer Research 04/2004; 64(6):1972-4. · 7.86 Impact Factor
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ABSTRACT: Objective: To express nitroreductase gene NOR1 in Escherichia coli and to purify the expressed protein in order to get the polyclonal antibody of NOR1. Methods: The full length of NOR1 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and digested withBamHI andXhoI restriction endonucleases. The plasmid pGEX-4T-2 was also digested withBamHI andXhoI, then the NOR1, gene was inserted into vector pGEX-4T-2. The recombinant expression vector pGEX-4T-2/NOR1, was identified by sequencing and restriction enzymes digestion. E.coli Jm105 transformed with the recombinant plasmid was
induced by IPTG to express the GST fusion protein. The purified targeted protein obtained by affinity chromatography was used
to immunize New Zealand rabbits to acquire antiserum. Antiserum was analyzed with immunoblot. Results: The 1.25 kb NOR1 gene was successfully isolated. After induction, a new anticipated protein of 74 kDa appeared on sodium dodecylsulfate polyacrylamide
(SDS-PAGE). The result was confirmed by Western blot analysis, and the purified targeted protein was obtained by affinity
chromatography. The titer of antiserum was 1:8. Conclusion: A high level of expression of GST-NOR1, is obtained in JM 105, and its antiserum can be prepared successfully.
Chinese Journal of Cancer Research 02/2004; 16(1):11-14. · 0.18 Impact Factor
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Zhao-Yang Zeng,
Wei Xiong,
Fang Xiong,
Shou-Rong Shen,
Xiao-Ling Li,
Wei-Fang Li, Rong Wang,
Bing-Yi Xiao,
Song-Qing Fan,
He Huang,
Ming Zhou,
Gui-Yuan Li
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ABSTRACT: To get genotype and allele frequency distributions of seven short tandem repeat (STR) loci of chromosome 9p,D9S288,D9S157,D9S1748,D9S171,D9S161,D9S1817 and D9S1805 in Chinese Han population in Hunan area,blood samples were collected from the random Han individual in Hunan and the whole genomic DNA was extracted.STR loci were amplified by multiplex-PCR technique and genotyped by ABI 377 sequencer.Seventy-five alleles were detected,with frequencies ranging from 0.002 to 0.800,and constituted 243 genotypes. All the seven loci met Hardy-Weinberg equilibrium. The statistical analysis of seven STR loci showed H(heterozygosity) ranging from 0.347 to 0.844,DP(discrimination power) ranging from 0.346 to 0.841,PPE(probabilities of paternity exclusion) ranging from 0.308 to 0.738 and PIC(polymorphic information content) ranging from 0.328 to 0.822. The result indicated that there was a significant difference between Han ethnic group and the white and the black.
Hereditas (Beijing) 10/2003; 25(5):543-8.
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Wei Xiong,
Zhao-yang Zeng,
Fang Xiong,
Shou-rong Shen,
Xiao-ling Li,
Wei-fang Li, Rong Wang,
Song-qing Fan,
Yi-xin Yang,
Hou-de Zhou,
Ming Zhou,
Gui-yuan Li
[show abstract]
[hide abstract]
ABSTRACT: To get the genotype and allele frequency distributions of 8 short tandem repeat (STR) loci on chromosome 3p (D3S1297, D3S1489, D3S1266, D3S1568, D3S1289, D3S1300, D3S1285 and D3S3681) in Chinese Han population in Hunan area.
Blood samples were collected from the random Han individuals in Hunan and the whole genomic DNA was extracted. STR loci were amplified by multiplex-PCR technique and genotyped by ABI 377 sequencer.
Ninety-one alleles were detected, with frequencies ranging from 0.002 to 0.431, and these alleles constituted 312 genotypes. All the 8 loci met Hardy-Weinberg equilibrium. The statistical analysis of 8 STR loci showed the heterozygosity (H) >or= 0.729, the discrimination power (DP) >or= 0.725, the probabilities of paternity exclusion (PPE) >or= 0.596, and the polymorphic information content (PIC >or= 0.682). The result indicated that there was a significant difference between Han ethnic group and the white and the black.
These results could serve as valuable data to enrich the Chinese genetic database and play an important role in Chinese population genetic and forensic medical application.
Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics 10/2003; 20(5):413-6.
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ABSTRACT: BRD7 was isolated through cDNA representational difference analysis (RDA) (GenBank accession No. AF152604). Previous studies showed that BRD7 gene was down-regulated in nasopharyngeal carcinoma (NPC) cells and tissues, and three cSNPs (coding-region single nucleotide polymorphisms) were found on BRD7. In addition, six BRD7-interacting proteins were identified by yeast two-hybrid system. To study the function of this gene on carcinogenesis in NPC, BRD7 gene was transfected into HNE1 cells low-expressed BRD7 by using liposomes and a stable cell line over-expressing BRD7 was established. After two-dimensional gel electrophoresis(2-DE), twenty differentially expressed proteins were identified by MALDI-TOF-MS including argininosuccinate lyase, metalloproteinase inhibitor-2 precursor, proteaseome activator28 beta subunit, thyroid transcriptional factor-1, cyclinH (MO15-associated protein), and so on. These differentially expressed proteins are related to cell cycling, transcription regulation, signaling pathway etc. Therefore, BRD7 may exert its functions by mediating differential expression of these proteins.
Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica 10/2003; 35(9):816-22.
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ABSTRACT: LRRC4 is a novel gene that the author has identified recently, which displayed significant downregulation in primary brain tumor biopsies. This study was designed to investigate if LRRC4 has the potential of suppressing brain tumor growth.
The full-length coding region of LRRC4 gene was subcloned into the expression vector pcDNA3.1, the recombinant was introduced into the glioblastoma cell line U251 by liposome transfection, and the U251 cells stably expressing LRRC4 gene were established by G418 selection. Furthermore, cell proliferation assay, soft agar assay, tumorigenesis assay were taken to examine the effect of LRRC4 expression on cell growth and tumor formation.
U251 cells stably expressing full-length coding region of LRRC4 were established by lipofection-mediated transfection and selected for further study. Compared with the nontransfected and vector-transfected cells, the cells transfected with LRRC4 cDNA exhibited a significant increase of expression of LRRC4 mRNA by Northern blot analysis. Further, when cell proliferation was followed over several days, the cells expressing the transfected LRRC4 cDNA grew more slowly than nontransfected cells. Consistently, the cells transfected with LRRC4 exhibited markedly lower colony formation rate. These clones were injected into athymic nude mice who was killed after 40 days and the tumor sizes were evaluated. Tumor volume in mice was significantly smaller in the group of cells stably transfected with LRRC4 cDNA than in the control.
LRRC4 gene may be transfected into the human glioblastoma cell line U251. The expression of LRRC4 in U251 cells may have the potential to suppress tumor cell growth and the tumorigenesis of U251 cell transplanted in nude mice.
Ai zheng = Aizheng = Chinese journal of cancer 10/2003; 22(9):897-902.
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ABSTRACT: Objective: To identify the relation between nasopharyngeal carcinoma and the human novel gene UBAP1, which is located in the
region of minimal heterozygosity deletion at 9pl3.2 and down-expressed in NPC. Methods: Five single nucleotide polymorphisms
(SNPs) within UBAP1 gene were analysed by sequencing in 105 NPC patients and 183 control subjects which matched to the NPC
cases on age, sex and residence. Results: Significant association was found between NPC with one SNP mark (rslO49557), which
is located at 3′ non-region of UBAP1 gene; the relative risk of this SNP mark is 1.64 (genotype GG) and 1.31 (genotype CG).
Conclusion: The result has proved again that UBAP1 gene may play a certain role in the occurrence and development of nasopharyngeal
carcinoma. The SNP mark rslO49557, considering its location, may influence the expression of UBAP1 gene.
Chinese Journal of Cancer Research 08/2003; 15(3):157-160. · 0.18 Impact Factor
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ABSTRACT: Malignant cells are often associated with aberrant expression or localization of connexins. Expression of Cx26, Cx32, Cx43, and Cx45 genes were studied to gain their expression profile in tissue microarrayers consisting of various carcinomas and to elucidate their function in carcinogenesis.
Basing on the principle of constructing tissue microarrays, the authors take use of self-made tissue arrayer to construct tissue microarrays for the different kinds of cancer specimens. Meanwhile, expression of connexin(Cx) genes, such as Cx32, Cx26, Cx43, and Cx45 in the tissue microarrays were detected in 292 cases of tumor tissues and 89 paratumor tissues by immunohistochemistry.
(1) Three kinds of tissue microarrays consisting samples from 20 different carcinomas were constructed successfully, which contained 360, 200, or 100 spots in each receptive paraffin block with tissue cylinder of 0.75mm or 0.6mm diameter respectively.(2) The expression profiles of Cx32, Cx26, Cx43, and Cx45 in 20 kinds of carcinomas and their related adjacent-cancer tissues were gained, which were widely low expressed in carcinomas especially in nasopharyngeal carcinomas. Expression of Cx26, Cx32, Cx43, and Cx45 in carcinomas such as carcinomas of colon and recta, hepatocellular carcinomas, gastric carcinomas, lung carcinomas and thyroid cancers were significantly lower than their related adjacent-cancer tissues.
Low expression of Cx26, Cx32, Cx43, and Cx45 might play crucial roles in carcinogenesis of many kinds of carcinomas. Tissue microarrays consisting of many different kinds of carcinomas provide a new high-throughout tool for rapid and comprehensive molecular expression profiles of carcinomas, such as connexin genes.
Ai zheng = Aizheng = Chinese journal of cancer 08/2003; 22(7):686-90.
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Wei Xiong,
Zhao-yang Zeng,
Shou-rong Shen,
Xiao-ling Li,
Wei-fang Li, Rong Wang,
Fang Xiong,
Cong Peng,
Qiu-hong Zhang,
Ming Zhou,
Yi-xin Yang,
Hou-de Zhou,
He Huang,
Gui-yuan Li
[show abstract]
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ABSTRACT: To investigate the relationship of nasopharyngeal carcinoma (NPC) with the high frequency allele imbalance locus D6S1581, and the NPC associated gene FBXO30 which is located near D6S1581.
Genescan was used to genotype D6S1581 of 12 NPC pedigrees, 85 sporadic NPC patients and 181 normal volunteers. Then parametric/nonparametric linkage analysis and association analysis were performed.
D6S1581 was linked with NPC, a Lod score of 2.611436 (P=0.00245) was obtained, and a significant difference in allele frequency was observed between familial NPC and control (P<0.005).
These results suggest that D6S1581 is highly associated with NPC, and there may be one or more NPC associated genes near D6S1581, including FBXO30.
Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics 08/2003; 20(4):311-4.
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Wei Xiong,
Zhao-yang Zeng,
Shou-rong Shen,
Xiao-ling Li,
Hong-bin Lu,
Juan-juan Xiang,
Xin-min Nie,
Shi-guo Zhu,
Wei-fang Li, Rong Wang,
Lin He,
Gui-yuan Li
[show abstract]
[hide abstract]
ABSTRACT: To search novel SNPs in exons and regulatory regions of CDKN2A and two novel putative tumor suppressor genes NGX6 and UBAP1, which all reside on chromosome 9p21-22.
The exons and regulatory regions of those genes were amplified and sequenced in 96 subjects.
Two novel SNPs were found, one resides on the sixth exon of UBAP1 gene and the other on the fourth exon of CDKN2A gene. Two novel SNPs were submitted to the dbSNP database, and their access ID are rs3135929 and rs3088440. The polymorphic information contents of them are 0.102 and 0.213 respectively. There is linkage equilibrium between them, and the polymorphic information content of their haplotype is 0.302, higher than any of them individually.
The polymorphic information content can be improved by using haplotype analysis of several SNPs.
Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics 07/2003; 20(3):203-6.
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ABSTRACT: NOR(1)is a good candidate of tumor suppressor/susceptibility gene associated with nasopharyngeal carcinoma. This study was designed to construct the prokaryotic expression vector and to investigate the expression of nitroreductase gene NOR(1)in Escherichia coli and to purify expressed product.
Total RNA was subtracted from normal nasopharyngeal carcinoma tissue. The full length of NOR(1)gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and digested with BamHIand XhoI restriction endonucleases. The plasmid pGEX-4T-2 was also digested with BamHI and XhoI,then the NOR(1)gene was inserted into vector pGEX-4T-2. The recombinant expression vector pGEX-4T-2/NOR(1)was identified by sequencing and digested with restriction enzymes. E.coli Jm105 transformed with the recombinant plasmid was induced by IPTG to express GST fusion protein. The result was confirmed by Western blot analysis and the purified targeted protein was obtained by affinity chromatography.
The 1.25kb NOR(1)gene was successfully isolated. After induction, a new anticipated protein of 74 kDa appeared on sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). It existed not only in supernatant but also in precipitation of broken bacteria. The result was confirmed by Western blot analysis,and the purified targeted protein was obtained by affinity chromatography.
The successes in construction of expression vector of NOR(1), expression and purification of GST/NOR(1)fusion protein make it possible to prepare for the polyantibodies for NOR(1).
Ai zheng = Aizheng = Chinese journal of cancer 03/2003; 22(2):136-9.
