-
[show abstract]
[hide abstract]
ABSTRACT: The human oncogene B-cell-specific Moloney murine leukemia virus integration site 1 (BMI-1) is a member of the mammalian Polycomb group family. The overexpression of BMI-1 is associated with human malignancies. In this study, the effects of knockdown of BMI-1 by shRNA-mediated RNA interference on cell cycle and possible downstream targets in human cervical adenocarcinoma HeLa cells were investigated. As a result, when the shRNA plasmid was stably introduced into the cell line, the mRNA and protein of BMI-1 were specifically down-regulated, and the cells increased in the phase of G1 and cells in S phase significantly decreased by flow cytometric analysis; the knockdown of BMI-1 expression could lead to significant up-regulation of p16INK4a, HOXA9 and HOXC13 mRNA expression, but hTERT and HOXB4 mRNA expression did not change significantly. In conclusion, RNAi-mediated knockdown of BMI-1 expression can induce cell-cycle arrest and up-regulate p16INK4a, HOXA9 and HOXC13 in HeLa cells. Our results suggest that targeting BMI-1 might be a therapeutic potential for the treatment of cancer.
Medical Oncology 12/2011; 28(4):1201-9. · 2.14 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Asthma is a common polygenic disease, caused by complex interactions between multiple genes and environmental factors. Study of the gene-gene interactions would contribute to a new insight into the pathogenesis and therapeutics of asthma.
To evaluate the single and combined associations of eight single-nucleotide polymorphisms loci in five candidate genes with the development of asthma in Chinese children.
We examined eight single-nucleotide polymorphisms (SNPs) in five key asthma susceptibility genes and performed single SNP association study, haplotype analysis, and gene-gene interactions analysis in 479 Chinese children, including 252 asthmatic subjects and 227 healthy controls. Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Haplotype analysis was detected by SHEsis software. Gene-gene interactions were tested using the multifactor dimensionality reduction (MDR) method.
There were significant differences of interleukin (IL)-13 R130Q and IL-13 C1923T in genotype and allele frequency distributions between the asthmatic group and control group. Furthermore, the A allele of IL-13 R130Q and the T allele of IL-13 C1923T were significantly associated with increased risk of asthma (odds ratio [OR] = 1.59, 95% confidence interval [CI] 1.20-2.09, p = .0010; OR = 1.57, 95% CI 1.19-2.08, p = .0014, respectively). By haplotype analysis, the C-G and T-A haplotypes consisting of IL-13 C1923T and IL-13 R130Q and the G-A and A-A haplotypes consisting of IL-4Ralpha I75V and IL-4Ralpha Q576R were significantly associated with asthma (p < .05). Using MDR, the authors detected significant gene-gene interactions with a best six-locus model among IL-4 -C33T, IL-13 R130Q, IL-4Ralpha I75V, IL-4Ralpha Q576R, STAT6 C2892T, and CD14 -C159T on the risk of asthma (OR = 4.43, 95% CI 1.30-15.04, p < .001, by 1000-fold permutation test).
These data suggest that genetic variants in the IL-13 gene may play an important role in the development of pediatric asthma in Middle China. In addition, the significant gene-gene interactions among IL-4 -C33T, IL-13 R130Q, IL-4Ralpha I75V, IL-4Ralpha Q576R, STAT6 C2892T, and CD14 -C159T may increase an individual's susceptibility to asthma and contribute to the pathogenesis of asthma.
Journal of Asthma 04/2010; 47(3):238-44. · 1.52 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: There are limited prospective data on clopidogrel resistance and clinical outcome of patients with selective coronary drug-eluting stent (DES) implantation.
To investigate whether clopidogrel resistance is associated with long-term thrombotic events in patients with selective coronary DES implantation.
A total of 154 patients who underwent selective percutaneous coronary intervention (PCI) with DES were enrolled in this study. Platelet aggregation was measured using light transmittance aggregometry (LTA) before clopidogrel administration (baseline) and 24 hours after loading with clopidogrel 300 mg. Clopidogrel resistance was defined as ≤10% absolute difference between baseline aggregation and post-administration aggregation. All patients who received the same anti-platelet treatment were followed up for 1 year after discharge for the incidence of a composite endpoint consisting of cardiovascular death, myocardial infarction (MI) and revascularization, and secondly for the incidence of stent thrombosis.
The incidence of clopidogrel resistance is 20.28% in our study population. Patients who are complicated by diabetes mellitus, smoke, or have a higher body mass index (BMI) tend to have clopidogrel resistance. Patients in the clopidogrel-resistant group have significantly higher incidences of composite endpoints (21.88% vs 4.92%; p = 0.006) and stent thrombosis (12.5% vs 1.64%; p = 0.017) than patients in the clopidogrel-response group during 1-year follow-up.
Diabetes, smoking, and high BMI are associated with clopidogrel resistance, and clopidogrel resistance indicates an increased risk of long-term thrombotic events in patients implanted with DES.
Drugs in R&D. 01/2010; 10(4):219-24.
-
[show abstract]
[hide abstract]
ABSTRACT: The accurate assessment of a proto-oncogene, human epidermal growth factor receptor-2 gene (HER-2), is extremely important for the therapy and prognosis of breast cancer. Currently, immunohistochemistry (IHC) is the method widely used for the detection of HER-2 protein. Fluorescence in situ hybridization (FISH) has been suggested to be a golden standard assay for HER-2 amplification. This study examined the expression and amplification of HER-2 in paraffin-embedded sections of breast cancer tissues, and compared the two methods on the measurement of HER-2 status. HER-2 gene and protein were determined in breast cancer samples from 52 Chinese women by FISH and IHC respectively. The findings indicated that the HER-2 gene amplification was found in 18 cases (34.6%) by FISH and the HER-2 protein over-expression (score 3+) in 15 cases (28.8%) by IHC. Immunohistochemically, 28.6% of the cases scored as 2+ and 93.3% of the cases scored as 3+ were HER-2-positive by FISH. There was a significant correlation between the HER-2 gene amplification and HER-2 protein over-expression in breast cancer (P<0.005). No correlation was noted between the HER-2 gene amplification and any of the clinicopathological parameters examined, including age, menopausal status, menarche age, tumor size, histological tumor type, histological grade, lymph node status, and the expression of ER and PR. It was concluded that the detection of HER-2 gene amplification in breast cancer by FISH is valuable and can compare with HER-2 protein detection by IHC.
Journal of Huazhong University of Science and Technology 06/2009; 29(3):354-8. · 0.38 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the distribution characteristics and linkage disequilibrium of T cell immunoglobulin domain and mucin domain protein 4 (TIM4) promoter polymorphisms in asthma patients of Chinese Han population, the promoter region of TIM4 was re-sequenced by PCR-sequencing, and linkage disequilibrium was analyzed by SHEsis software. Four single nucleotide polymorphisms (SNPs) in the promoter region of TIM4 were detected, including two new SNPs (at positions-1609,-153) and two reported SNPs (rs6874202, rs6882076). The frequency distribution of rs6882076 was different among different races (P<0.05). In addition, linkage disequilibrium among the SNPs of the promoter region of TIM4 was found and GGTG was the predominant haplotype. There were four SNPs in the promoter region of TIM4 in asthma patients of Chinese Han population, which were in linkage disequilibrium.
Journal of Huazhong University of Science and Technology 08/2008; 28(4):447-50. · 0.38 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To observe the inhibitory effect of cyclin-dependent kinase inhibitor p21 on regulation of survivin transcription in human liver cancer HepG cells, and explore the related mechanisms.
Doxorubicin (DOX) was used to treat HepG cells. Eukaryotic vector pEGFP-C2-p21 was transfected into HepG cells by lipofectamine and positive clones were screened out by G418. The mRNA expression of p21, p53 and survivin were detected by real-time fluorescent quantitative polymerase chain reaction (RQ-PCR). Flow cytometry was used to determine the cell cycle phases, and reverse transcription polymerase chain reaction (RT-PCR) was used to measure the levels of E2F-1 or p300.
After treatment with DOX, the expression of p53 and p21 was increased, whereas that of survivin was reduced during 24 hours of the treatment. After transfection the p21 level was 2100.1-fold or 980.9-fold enhanced in comparison with that in HepG2 cells or HepG2-pEGFP cells. Survivin level was markedly down-regulated to 0.5% or 0.6% relative to that in the other two groups, nevertheless, significant p53 changes were not observed. Overexpression of p21 resulted in G1/G0 phase arrest (F = 31.59, P < 0.01), meanwhile, E2F-1 mRNA or p300 mRNA were less expressed compared with that in the other controls (F(E2F-1) = 125.28, P < 0.05; Fp300 = 46.01, P < 0.01).
p21 could be a potential mediator of survivin suppression at transcription level in HepG2 cells, which might be through the block at G1/G0 phase and down-regulation of transcription factors E2F-1 and p300.
Zhonghua zhong liu za zhi [Chinese journal of oncology] 08/2008; 30(8):583-7.
-
[show abstract]
[hide abstract]
ABSTRACT: The effect of cyclin-dependent kinase inhibitors Cip1/Waf1 (p21) on regulatory expression of survivin transcription in human hepatocellular carcinoma cell HepG2 was observed and the related mechanisms explored. Doxorubicin (DOX) was used to treat HepG2. Eukaryotic vector pEGFP-C2-p21 was transfected into HepG2 by lipofectamine and positive clones were screened out by G418. The mRNA expression of p21 and survivin was detected by real-time fluorescent quantitative polymerase chain reaction (RQ-PCR). Flow cytometry was used to examine the cell cycle, and reverse transcription polymerase chain reaction (RT-PCR) was used to measure the levels of E2F-1 and p300. The results showed that: (1) After treatment with DOX, the expression of p21 was increased, whereas that of survivin was reduced during 24 h of treatment; (2) After transfection of pEGFP-C2-p21 into HepG2, p21 level was significantly enhanced to 2100.11-folds or 980.89-folds in comparison to HepG2 or HepG2-C2 group, and survivin level was markedly down-regulated to 0.54% or 0.59% relative to the control groups; (3) Overexpressed p21 resulted in G1/G0 phase arrest (F=31.59, P<0.01), meanwhile E2F-1 mRNA and p300 mRNA were reduced as compared with those of controls (F(E2F-1)=125.28, P<0.05; F(p300)=46.01, P<0.01). It was suggested that p21 could be a potential mediator of survivin suppression at transcription level in HepG2 cell, which might be through the block at G1/G0 phase and down-regulation of transcription factors E2F-1 and p300.
Journal of Huazhong University of Science and Technology 06/2008; 28(3):308-13. · 0.38 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Chemotherapy protocols using adriamycin (ADM) is a standard treatment for hepatoblastoma, but the treatment results became unsatisfied because of drug resistance. Recently, ADM combined gene therapy is a developing alternative treatment for hepatoblastoma. This study was to investigate the effect of ADM combined human p21CIP1 transfection on the proliferation of hepatoblastoma cell line HepG2.
HepG2 cells were divided into empty control group (no treatment), ADM group (treated with 0.5 microg/mL ADM), blank control group (transfected with blank plasmid pcDNA3), p21 group (transfected with plasmid pcDNA3-p21), and combination group (ADM treatment plus p21 transfection). The proliferation of HepG2 cells was observed by MTT assay. The mRNA levels of p21 and survivin were detected by real-time polymerase chain reaction (PCR).
After transfection, the mRNA level of p21 in p21 group was increased by 155 folds of that in empty control group (P<0.05). p21 inhibited the proliferation of HepG2 cells at Day 3 and Day 4 after transfection (P<0.01). The proliferation inhibition rate was significantly higher in combination group than in ADM group and p21 group (43.92% vs. 32.97% and 35.77% at Day 3, P<0.01; 59.86% vs. 39.35% and 40.96% at Day 4, P<0.01; 51.81% vs. 33.91% and 10.68% at Day 5, P<0.01). This effect was enhanced along with the increasing time of co-treatment from Day 1 to Day 4 (r=0.91, P<0.05), and it was obvious at Day 4 (Q =1.07). The mRNA level of survivin was significantly lower in combination group than in p21 group and ADM group (P<0.01).
p21 gene transfection plus ADM can inhibit the proliferation of HepG2 cells and down-regulate the level of survivin mRNA, thus may be a potential therapeutic strategy against human hepatoblastoma.
Ai zheng = Aizheng = Chinese journal of cancer 05/2008; 27(5):476-81.
-
[show abstract]
[hide abstract]
ABSTRACT: In order to investigate peptide mimics of carbohydrate blood group A antigen, a phage display 12-mer peptide library was screened with a monoclonal antibody against blood group A antigen, NaM87-1F6. The antibody-binding properties of the selected phage peptides were evaluated by phage ELISA and phage capture assay. The peptides were co-expressed as glutathione S-transferase (GST) fusion proteins. RBC agglutination inhibition assay was performed to assess the natural blood group A antigen-mimicking ability of the fusion proteins. The results showed that seven phage clones selected bound to NaM87-1F6 specifically, among which, 6 clones bore the same peptide sequence, EYWYCGMNRTGC and another harbored a different one QIWYERTLPFTF. The two peptides were successfully expressed at the N terminal of GST protein. Both of the fusion proteins inhibited the RBC agglutination mediated by anti-A serum in a concentration-dependent manner. These results suggested that the fusion proteins based on the selected peptides could mimic the blood group A antigen and might be used as anti-A antibody-adsorbing materials when immunoabsorption was applied in ABO incompatible transplantation.
Journal of Huazhong University of Science and Technology 05/2008; 28(2):222-6. · 0.38 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the single nucleotide polymorphisms (SNPs) of -1516G/T in the promoter region and 4259G/T in the exon-3 region of the T cells immunoglobulin mucin-3 (TIM-3) and their linkage disequilibrium, and therefore to detect their haplotype relationship with allergic asthma of the Han population from Hubei province of China.
The two polymorphisms were detected with allelic specific polymerase chain reaction (ASPCR). In the 175 asthmatic subjects and in the 202 healthy controls collected from June, 2004 to October 2007 in the Han population from Hubei province. The genotype and allele frequencies, the D' value between the two SNPs sites, the haplotype and their frequencies were calculated and analyzed, respectively.
The genotype frequencies of GG, GT and TT in -1516G/T polymorphism of TIM-3 gene were 82.7% (167/202), 17.3% (35/202), 0 (0/202) respectively in the 202 controls, and 82.9% (145/175), 17.1% (30/175), 0 (0/175) respectively in the 175 asthmatic subjects. The genotype frequencies of GG, GT and TT in 4259G/T polymorphism of TIM-3 gene were 0.5% (1/202), 2.5% (5/202), 97.0% (196/202) respectively in the 202 controls and 0.6% (1/175), 5.7% (10/175), 93.7% (164/175) respectively in the 175 asthmatic subjects. The control group: D' = 1.0, the asthma group D' = 0. 9. The 3 haplotypes were G-G, G-T, T-T in the Han population from Hubei province of China, and their haplotype frequencies were distributed similarly in asthma 3.4% (12/350), 88.0% (308/ 350), 8.6% (30/350) and in the controls 1.7% (7/404), 89.6% (362/404), 8.7% (35/404). None of these differences were statistically significant (chi2 = 2.15, 0.47, 0.003 respectively, all P > 0.05).
There are strong linkage disequilibrium between the two SNPs sites in TIM-3 gene in Han population from Hubei province, but the haplotypes G-G, G-T and T-T are not associated with asthma susceptibility of this population. We cannot exclude the possibility that the haplotypes of TIM-3 may be associated with asthma susceptibility in other ethnic populations or the susceptibility to other atopic diseases and autoimmunity diseases.
Zhonghua jie he he hu xi za zhi = Zhonghua jiehe he huxi zazhi = Chinese journal of tuberculosis and respiratory diseases 04/2008; 31(3):196-200.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the frequencies of -1516,-574 and 4259 single nucleotide polymorphisms (SNPs) of T cells immunoglobulin mucin -3(TIM-3) gene in Hubei population and address the question whether they are in linkage disequilibrium(LD) .
Genotypes and allele frequencies of TIM-3 gene were examined by allele-specific polymerase chain reaction (AS-PCR) methods in 147 healthy Hubei Han individuals. Hardy-Weinberg equilibrium and Two-point LD analyses and haplotype frequencies were evaluated with Arlequin v3.1 software.
The allele frequencies of the 3 SNPs were in agreement with Hardy-Weinberg equilibrium. Minor allelic frequencies of TIM-3 -1516G/T,-574T/G and 4259G/T were 8.5%,1.0% and 2.0%,respectively. The dominant haplotypes comprising the three loci were G-G-G(2.0%),G-G-T(88.4%), T-G-T(8.5%) and G-T-T(1.0%). LD analyses revealed that all of the coefficient of linkage disequilibrium (D') were 1.
The -1516,-574 and 4259 loci of TIM-3 gene are in complete linkage disequilibrium. Our study has provided population genetic data on TIM-3 gene in Chinese Hubei Han population and a basis for searching immune-mediated disease-related TIM-3 haplotype.
Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics 03/2008; 25(1):101-4.
-
[show abstract]
[hide abstract]
ABSTRACT: This study was purposed to construct and identify the mammalian expression vector of pEGFP-BMI-1 and to detect whether it could express in human cervix cancer cell line HeLa. The cDNA fragment of BMI-1 obtained by RT-PCR was inserted into pEGFP-N1. The recombinant plasmid was confirmed by restriction enzyme digestion, PCR and DNA sequencing. pEGFP-BMI-1 was transfected into HeLa cells with lipofectamine 2000. The expression of pEGFP-BMI-1 was determined by EGFP fluorescence and Western blot analysis. SYBR Green I real-time RT-PCR was used to quantitate P16INK4a mRNA. The results showed that the correct construction of the recombinant plasmid pEGFP-BMI-1 has been shown by restriction enzyme digestion, PCR and DNA sequencing. pEGFP-BMI-1 could express BMI-1-EGFP fusion protein in HeLa cells. Real-time RT-PCR showed that P16INK4a mRNA expression was reduced to 9.2%. It is concluded that the vector of pEGFP-BMI-1 has been successfully constructed and it can be expressed in HeLa cells. This work has laid foundations for further study on biological functions and potential application of BMI-1.
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology 11/2007; 15(5):1056-60.
-
[show abstract]
[hide abstract]
ABSTRACT: A plasmid carrying DNA to be transcribed into a small interfering RNA against transketolase-like-1 mRNA was constructed and transfected into a human colon cancer cell line. The mRNA expression of transketolase gene family in the human colon cell line was determined by real-time polymerase chain reaction. The effect of anti-transketolase-like-1 small interfering RNA on cell proliferation and cell cycle in the human colon cancer cell line cells was detected by flow cytometry and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide. The transketolase-like-1 gene was significantly downregulated in human colon cancer cell line cells transfected with small interfering RNA transketolase-like-1 constructs compared with the cells transfected with control vector and the cells without transfection. In addition, the anti-transketolase-like-1 small interfering RNA construct significantly decreased the level of transketolase in the transfected human colon cancer cell line cells, arrested them in G0/G1 phase and substantially inhibited cell proliferation. No significant difference was found in the other two genes (transketolase and transketolase-like-2 genes) between the transfected human colon cancer cell line cells and the controls (P>0.05). Our data demonstrated that the transketolase-like-1 gene plays an important role in total transketolase activity and in the cell proliferation of human colon cancer. Transketolase-like-1 may serve as a target for novel anticancer therapies.
Anti-Cancer Drugs 04/2007; 18(4):427-33. · 2.41 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Beta-catenin plays a central role in Wnt signaling pathway. The aberrant localization of beta-catenin in nucleus causes the transcription of down-stream target genes, which is the pathogenesy of some solid tumours. As the expression of adheren junction on hemopoietic cells is very low, there are a few studies on beta-catenin expression in leukaemia. This study was aimed to investigate beta-catenin localization and beta-catenin mRNA expression levels in 4 leukemia cell lines so as to explore a new oncogenic mechanism and to find out a new therapeutic target. The beta-catenin localization in leukemia cell lines was detected by immunocytochemistry, the beta-catenin mRNA expression level was assayed by real-time quantitative RT-PCR. The results showed that there was aberrant localization of beta-catenin in Jurkat and Thp-1, and beta-catenin mRNA expression level was not increased in these two cell lines, however, the mRNA expression levels of Jurkat and Thp-1 were lower than those of Daudi and K562. The beta-catenin mRNA expression level was not correlated with beta-catenin aberrant localization in these 4 cell lines. It is concluded that the aberrant localization of beta-catenin may play a role in the development of some leukemia, and the mechanism resulting in beta-catenin aberrant localization not take place at transcription level.
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology 01/2007; 14(6):1096-100.
-
[show abstract]
[hide abstract]
ABSTRACT: In this study, we examined the effects of aspirin on the growth rates, subcellar distribution of beta-catenin protein, the expression of beta-catenin/TCF signaling pathway target gene cyclinD1 mRNA, and cell cycle of Jurkat cell line (Human T-acute lymphoblastic leukemia). Our results showed that the treatment with aspirin inhibited the growth of Jurkat cell line. Jurkat cells treated with 3 mmol/L of aspirin could significantly decrease nuclear localization of beta-catenin, and at 5 mmol/L of aspirin, the nuclear localization of beta-catenin was undetectable. QRT-PCR showed that the target gene cyclinD1 mRNA expression was gradually decreased with the dosage of aspirin. Aspirin induced G0/G1 cell cycle arrest in Jurkat cells. We are led to conclude that aspirin acts through beta-catenin-independent mechanisms. The effects of aspirin include down-regulation of beta-catenin nuclear localization and G0/G1 cell cycle arrest, which might serve as a means of growth inhibition in aspirin-treated human Jurkat cell line.
Journal of Huazhong University of Science and Technology 02/2006; 26(6):731-4. · 0.38 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Upregulation of hTERT mRNA plays an important role during the occurrence and development of laryngeal squamous cell carcinoma. Most hTERT mRNA in plasma from patients with laryngeal squamous cell carcinoma is derived from tumour cells; moreover, the determination of plasma hTERT mRNA contributes to tumour diagnosis and the observation of curative effect.
To establish a real-time fluorescent reverse transcriptase polymerase chain reaction (RT-PCR) and to quantitate the level of human telomerase reverse transcriptase (hTERT) mRNA in carcinoma tissue and plasma from patients with laryngeal squamous cell carcinoma. We also wished to evaluate the role that hTERT mRNA expression plays during the occurrence and development of laryngeal squamous cell carcinoma, to probe the correlation between the expression level and the clinical and pathological parameters and to investigate the value of the determination of plasma hTERT mRNA in tumour diagnosis and the observation of curative effect.
A real-time fluorescent RT-PCR and a Lightcycler PCR system were used to quantitate the expression level of hTERT mRNA.
The expression levels of hTERT mRNA (NhTERT) from laryngeal squamous cell carcinoma tissue and corresponding adjacent non-cancerous tissue were 62.6 +/- 21.7 and 3.5 +/- 1.9, respectively. NhTERT was significantly elevated in laryngeal squamous cell carcinoma tissue, rising to 17.9-fold on average, but there was no significant correlation between NhTERT and either tumour location, differentiation degree, T grade or N grade. For healthy examinees, NhTERT in plasma was 1.3 +/- 0.9, compared to 13.1 +/- 9.4 and 9.3 +/- 5.8 in patients with laryngeal squamous cell carcinoma examined before and 2 days after surgery, respectively. Compared to healthy examinees, NhTERT in plasma from patients with laryngeal squamous cell carcinoma was significantly elevated; moreover, 2 days after surgery, NhTERT in plasma had decreased significantly in these patients.
Acta Oto-Laryngologica 05/2005; 125(4):409-14. · 1.08 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To detect the expression of telomerase subunits (human telomerase reverse transcriptase, human telomerase associated protein 1 and human telomerase RNA) in gastric cancer and to examine the role that different telomerase subunits play in the gastric carcinogenesis, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect telomerase subunits messenger RNA in 24 samples of gastric cancer and corresponding non-cancerous tissue. The results showed that the positive rate of hTERT mRNA from gastric cancer and corresponding non-cancerous tissues was 100% and 25%, respectively. The former was significantly higher than the latter (chi2 = 26.4, P < 0.01). The positive rate of hTEP1 mRNA from gastric cancer and corresponding non-cancerous tissues was 100% and 91.7%, respectively and no significant difference was found between them (chi2 = 2.1, P > 0.05). The positive rates of hTR for gastric cancer and corresponding non-cancerous tissues were both 100% and no significant difference existed between them. It is concluded that in contrast to hTEP1 and hTR, the up-regulation of hTERT mRNA expression may play a more important role in the development of gastric cancer.
Journal of Huazhong University of Science and Technology 02/2005; 25(6):741-3. · 0.38 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To set up a real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay, to detect human telomerase reverse transcriptase (hTERT) messenger RNA in gastric carcinomas, and to evaluate quantitative determination of hTERT mRNA in the diagnostic value of gastric carcinomas, and to analyze the correlation between the expression level of hTERT mRNA and clinicopathological parameters in patients with gastric cancer.
A real-time quantitative RT-PCR (RQ-PCR) based on TaqMan fluorescence methodology and the LightCycler system was used to quantify the full range of hTERT mRNA copy numbers in 35 samples of gastric carcinomas and corresponding adjacent non-cancerous tissues. The normalized hTERT (NhTERT) was standardized by quantifying the number of GAPDH transcripts as internal control and expressed as 100X (hTERT/GAPDH) ratio. Variables were analyzed by the Student's t-test, chi2 test and Fisher's exact test.
NhTERT from gastric carcinomas and corresponding adjacent non-cancerous tissues was 6.27+/-0.89 and 0.93+/-0.18, respectively (t = 12.76, P<0.001). There was no significant association between gastric cancer hTERT mRNA expression level and patient's age, gender, tumor size, location and stage (pTNM), but a significant correlation was found between hTERT mRNA expression level in gastric carcinomas and the degree of differentiation.
Quantitative determination of hTERT mRNA by RQ-PCR is a rapid and sensitive method. hTERT might be a potential biomarker for the early detection of gastric cancer.
World Journal of Gastroenterology 12/2004; 10(23):3514-7. · 2.47 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Telomerase activity and the expression of telomerase subunits (for example, telomerase reverse transcriptase and telomerase associated protein 1 and telomerase RNA component) of peripheral white blood cells were detected in the patients with acute myelogenous leukaemia (AML) and the correlation between telomerase activity and the expression of telomerase subunits was observed. In 94 peripheral white blood cells from 18 healthy volunteers and 76 patients with AML, including 31 AML at initial presentation, 24 at relapse and 21 at complete remission, the telomerase activity and telomerase subunits mRNA or RNA were detected by PCR-ELISA and RT-PCR respectively. The results showed that the positive rate of telomerase from patients with AML at initial presentation, at relapse and at complete remission was 74.1%, 79.2% and 4.8% respectively. The positive rate of telomerase reverse transcriptase mRNA from healthy volunteers, AML at initial presentation, AML at relapse and AML at complete remission was 5.6%, 80.6%, 83.3% and 9.5% respectively. The positive rate of telomerase associated protein 1 mRNA and telomerase RNA component in all samples were 100%. It was suggested that the up-regulation of telomerase activity and the expression of telomerase reverse transcriptase is correlated closely with the occurrence and relapse of AML, so telomerase activity and the expression of telomerase reverse transcriptase may be used to estimate the curative effect and predict relapse of AML. Moreover, the up-regulation of telomerase activity is correlated with the expression of telomerase reverse transcriptase significantly.
Journal of Huazhong University of Science and Technology 02/2004; 24(1):48-51. · 0.38 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the link between the polymorphism of -109 and Glu237 in the high-affinity IgE receptor beta (FcepsilonRIbeta) gene and susceptibility to allergic asthma in a Chinese population.
Blood samples from 216 allergic asthma patients and 198 age- and sex-matched controls were studied. A -109C/T and a coding variant Glu237Gly in FcepsilonRIbeta were detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).
The genotype frequencies were 0.403 for -109T/T, 0.491 for -109T/C and 0.106 for -109C/C in allergic asthma in a Chinese population. No significant difference in the distribution of -109C/T polymorphism was found between allergic asthma subjects and healthy controls, however, homozygosity for the -109T allele was associated with increased total plasma IgE levels in subjects with allergic asthma (F = 4.020, P < 0.05). The allele frequency of Gly237 in the patients and control was 0.236 and 0.136 respectively. There was a significant association between the Gly/Gly genotype and allergic asthma. Among allergic asthma patients Gly237 was significantly associated with high IgE levels.
These results suggest that the Gly237 variant of the FcepsilonRIbeta gene is involved in the development of allergic asthma. The -109C/T and Glu237Gly polymorphisms are two of the genetic factor identified thus far, which affect total plasma IgE levels of allergic asthma patients in a Chinese population.
Chinese medical journal 12/2003; 116(12):1875-8. · 0.86 Impact Factor
