1.Distribution of tap1 and tap2 loci allelic and genotypic frequencies in Chinese Hans.
Wu SHEN ; Tong SHEN ; Biao YIN ; Jie-Wei ZHENG ; Da-Zhuang LIU ; Ming-Liang FENG
Journal of Experimental Hematology 2009;17(3):777-781
This study was aimed to investigate the distribution and implication of tap1 (transporter associated with antigen processing) and tap2 loci allelic and genotypic frequencies. The distribution of tap1 and tap2 loci allelic and genotypic frequencies in 339 random samples of healthy Chinese Hans was analyzed by TaqMan PCR. Several genetic information about power of discrimination, cumulative DP, polymorphism information content, expected heterozygosity and observed heterozygosity were calculated. The results indicated that 5 tap1 alleles (tap1*0101, 020101, 020102, 0301 and 0401) and 4 tap2 alleles (tap2*0101, 0102, 0103 and 0201) were detected in all samples. 8 tap1 genotypes were found which account for 53.3% of the theoretic genotype and 6 tap2 genotypes were found which account for 60% of the theoretic genotype. The genotyping results of tap1 and tap2 both conform to the Hardy-Weinberg expectations (p > 0.05). Tap1*0101 (79.79%) and tap2*0101 (82.74%) are the most common alleles in Chinese Hans. It is concluded that tap1*0101 and tap2*0101 are most common alleles in Chinese Hans, tap1 and tap2 loci carry some power of individual discrimination and polymorphism information content. These two locl can be used for the research in the fields of human genetics, linkage analysis of genetic disease genes, paternity test and individual identification and so on.
ATP-Binding Cassette Sub-Family B Member 2
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ATP-Binding Cassette Transporters
;
genetics
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ATP-Binding Cassette, Sub-Family B, Member 3
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Alleles
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Asian Continental Ancestry Group
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genetics
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Gene Frequency
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Genotype
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Haplotypes
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Humans
2.Association of expression of antigen processing machinery components with HLA-I in cervical lesions.
Ayshamgul HASIM ; Song QING ; Xiao-chuan WU ; Lin GE ; Abulizi ABUDULA
Chinese Journal of Pathology 2010;39(10):703-704
ATP-Binding Cassette Sub-Family B Member 2
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ATP-Binding Cassette Transporters
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metabolism
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ATP-Binding Cassette, Sub-Family B, Member 3
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Calnexin
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metabolism
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Calreticulin
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metabolism
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Carcinoma, Squamous Cell
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metabolism
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pathology
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virology
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Cervical Intraepithelial Neoplasia
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metabolism
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pathology
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virology
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Cysteine Endopeptidases
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metabolism
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Female
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HLA-A Antigens
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metabolism
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Human papillomavirus 16
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isolation & purification
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Humans
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Lymphatic Metastasis
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Papillomavirus Infections
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Proteasome Endopeptidase Complex
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metabolism
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Uterine Cervical Neoplasms
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metabolism
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pathology
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virology
3.Relationship between TAP gene promoter methylation and cervical lesions with HPV infection in Uyghur women.
Guzali AIKUER ; Zhen JIAO ; Xiao-chuan WU ; Jun-qi MA ; Abulizi ABUDLA ; Ayshamgul HASIM
Chinese Journal of Pathology 2012;41(7):438-442
<b>OBJECTIVEb>To study the relationship between TAP (transporter associated with antigen processing) gene promoter regional methylation level and cervical lesions with HPV infection in Uyghur women.
<b>METHODSb>A specialized software was used to design specific primers of CpG island fragments of TAP1 and TAP2 gene promoter for PCR amplification, bisulfitemodified SiHa cancer cell DNA for PCR amplification, cloning and sequencing analysis to obtain the relevant information on the gene base sequence methylation of CpG sites. Seventy-eight fresh cervical tissue samples from Uyghur women with cervicitis (number = 15), cervical intraepithelial neoplasia (CIN, number = 30) and cervical squamous cell carcinoma (number = 33) were collected. The methylation level of TAP1 and TAP2 gene promoter regions was detected using MassArray DNA technology. HPV infection status was determined by HPV gene chips. The relationship between CpG-island methylation of gene promoter regions and HPV infection was then analyzed.
<b>RESULTSb>Each TAP1 and TAP2 gene corresponding target fragment contained 23 and 8 CpG sites. There were 5 and 8 CpG sites methylation occurred in SiHa cervical cancer cells genomic DNA respectively. The TAP1 methylation level increased steadily with the severity of cervical lesions. The methylation levels in cervical squamous cell carcinoma and CIN (0.048 ± 0.039 and 0.037 ± 0.026, respectively) were higher than that of normal cervical tissue (0.035 ± 0.029, P < 0.05). Although TAP2 gene methylation level also demonstrated similar changes, the difference however was not statistically significant (P > 0.05). HPV gene chip detected 13 HPV genotypes, with HPV16 infection rate being 66.7% (52/78). The methylated proportion of TAP1 positively correlated with HPV16 infection (χ(2) = 6.08, P = 0.039).
<b>CONCLUSIONb>TAP1 methylation is a remarkable phenomenon occurring in a range of cervical lesions and significantly associated with cervical HPV infection.
ATP-Binding Cassette Sub-Family B Member 2 ; ATP-Binding Cassette Transporters ; genetics ; ATP-Binding Cassette, Sub-Family B, Member 3 ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Carcinoma, Squamous Cell ; genetics ; virology ; Cervical Intraepithelial Neoplasia ; genetics ; virology ; CpG Islands ; genetics ; DNA Methylation ; Female ; Human papillomavirus 16 ; Humans ; Middle Aged ; Papillomavirus Infections ; Promoter Regions, Genetic ; Uterine Cervical Neoplasms ; genetics ; virology ; Uterine Cervicitis ; genetics ; virology
4.Association between polymorphisms of PSMB8, PSMB9 and TAP2 genes with rheumatoid arthritis in ethnic Han Chinese from Yunnan.
Liang YU ; Qin LI ; Jun LIN ; Juan YU ; Qian LI ; Wei YI ; Hao SUN ; Jia-you CHU ; Zhao-qing YANG
Chinese Journal of Medical Genetics 2013;30(2):222-226
<b>OBJECTIVEb>To assess the association between single nucleotide polymorphisms (SNPs) of PSMB8, PSMB9 and TAP2 genes and rheumatoid arthritis (RA) in ethnic Han Chinese from Yunnan.
<b>METHODSb>A case-control study was carried out using 177 RA patients and 288 healthy controls. Genotypes of rs2071543, rs55745125 and rs138635403 loci of PSMB8 gene, and rs17587 locus of PSMB9 gene were determined with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). And a polymerase chain reaction amplification refractory mutation system (ARMS-PCR) was used for typing rs2228396 locus of TAP2 gene. Genotypic and allelic frequencies were calculated. An Epi Info 7 software was used to calculate the Odds Ratio (OR) of above SNPs between the two groups.
<b>RESULTSb>Allelic and genotypic frequencies of rs138635403 and rs17587 loci have differed significantly between the two groups (P<0.05). The frequency of GG genotype for rs17587 locus was also higher in the RA group (0.672) compared with control group (0.524) (OR=1.862, 95%CI: 1.261-2.749).
<b>CONCLUSIONb>Genetic polymorphisms of rs17587 appeared to be associated with RA in ethnic Han Chinese from Yunnan.
ATP-Binding Cassette Transporters ; genetics ; ATP-Binding Cassette, Sub-Family B, Member 3 ; Arthritis, Rheumatoid ; genetics ; Case-Control Studies ; China ; ethnology ; Cysteine Endopeptidases ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Polymorphism, Single Nucleotide ; Proteasome Endopeptidase Complex ; genetics
5.Association Analysis of Proteasome Subunits and Transporter Associated with Antigen Processing on Chinese Patients with Parkinson's Disease.
Ming-Shu MO ; Wei HUANG ; Cong-Cong SUN ; Li-Min ZHANG ; Luan CEN ; You-Sheng XIAO ; Guo-Fei LI ; Xin-Ling YANG ; Shao-Gang QU ; Ping-Yi XU ;
Chinese Medical Journal 2016;129(9):1053-1058
<b>BACKGROUNDb>Proteasome subunits (PSMB) and transporter associated with antigen processing (TAP) loci are located in the human leukocyte antigen (HLA) Class II region play important roles in immune response and protein degradation in neurodegenerative diseases. This study aimed to explore the association between single nucleotide polymorphisms (SNPs) of PSMB and TAP and Parkinson's disease (PD).
<b>METHODSb>A case-control study was conducted by genotyping SNPs in PSMB8, PSMB9, TAP1, and TAP2 genes in the Chinese population. Subjects included 542 sporadic patients with PD and 674 healthy controls. Nine identified SNPs in PSMB8, PSMB9, TAP1, and TAP2 were genotyped through SNaPshot testing.
<b>RESULTSb>The stratified analysis of rs17587 was specially performed on gender. Data revealed that female patients carry a higher frequency of rs17587-G/G versus (A/A + G/A) compared with controls. But there was no significant difference with respect to the genotypic frequencies of the SNPs in PSMB8, TAP1, and TAP2 loci in PD patients.
<b>CONCLUSIONb>Chinese females carrying the rs17587-G/G genotype in PSMB9 may increase a higher risk for PD, but no linkage was found between other SNPs in HLA Class II region and PD.
ATP-Binding Cassette Sub-Family B Member 2 ; genetics ; ATP-Binding Cassette, Sub-Family B, Member 3 ; genetics ; Adult ; Aged ; Antigen Presentation ; Case-Control Studies ; Cysteine Endopeptidases ; genetics ; Female ; Humans ; Male ; Middle Aged ; Parkinson Disease ; genetics ; immunology ; Polymorphism, Single Nucleotide ; Proteasome Endopeptidase Complex ; genetics
6.K562 cell line resistance to nilotinib induced in vitro and preliminary investigation of its mechanisms.
Ji-shi WANG ; Chang YANG ; Qin FANG ; Si-xi WEI ; Cheng CHEN ; Yuan YANG ; Ya-ting WANG ; Xiu-ying HU ; Dan MA
Chinese Journal of Hematology 2012;33(11):906-910
<b>OBJECTIVEb>To establish a bcr-abl(+) cell line resistance to nilotinib, and to investigate the possible mechanisms of resistance.
<b>METHODSb>K562 cells were treated with gradually increasing concentrations of nilotinib to generate resistance cell line K562-RN. The folder of drug-resistance was evaluated by MTT assay. Cells apoptosis rate was detected by flow cytometry, the mRNA level of bcr-abl fusion gene by FISH, and the expression of apoptosis relative gene mRNA and protein (such as bcr-abl, HO-1, mdr1, Bcl-2 and caspase-3) by RQ-PCR and western blot.
<b>RESULTSb>The resistant cell line K562-RN was successfully established, with 2.01 fold resistant to nilotinib compared with K562 cell line \[the IC(50) value of nilotinib to K562 and K562-RN were (12.320 ± 1.720) µmol/L and (24.742 ± 2.310) µmol/L, respectively\]. It also had the cross resistance to adriamycin, homoharringtonine, etoposide and imatinib. Treated with different concentrations of nilotinib, cell apoptosis rate of K562-RN was significantly lower than that of K562 cells. The rate of bcr-abl gene positive cells was 92% in K562-RN by FISH assay. The mRNA and protein levels of bcr-abl, HO-1 and mdr1 expression up-regulated in K562-RN cells, while those of caspase-3 expression down-regulated, being significantly statistical difference when compared with K562 cells (P < 0.05).
<b>CONCLUSIONb>Human leukemic cell line resistance to nilotinib, K562-RN is established successfully by gradually increasing concentrations of drug. The mechanisms of resistance in K562-RN is probably associated with increasing expression of bcr-abl, HO-1, mdr1 and decreasing expression of caspase-3 mRNA and protein levels.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Caspase 3 ; metabolism ; Drug Resistance, Neoplasm ; Fusion Proteins, bcr-abl ; metabolism ; Gene Expression Regulation, Leukemic ; Heme Oxygenase-1 ; metabolism ; Humans ; K562 Cells ; drug effects ; Pyrimidines ; pharmacology
7.Expression of FLT3 internal tandem duplication in pediatric patients with acute myeloid leukemia and its correlation with multidrug resistance.
Jiang-Ning ZHAO ; Zhen-Hua QIAO ; Lian-Rong XU ; Quan-Yi LU ; Xiao-Qing NIU ; Peng ZHANG ; Zhao WANG
Journal of Experimental Hematology 2009;17(1):23-26
This study was aimed to investigate the expression of FLT3 internal tandem duplication (FLT3-ITD) in pediatric patients with acute myeloid leukemia (AML) and to analyse the clinical features of patients with mutations and the relation of FLT3-ITD with multidrug resistance gene 1 (mdr1). RT-PCR was used to determine the expressions of FIT3-ITD and mdr1 gene in bone marrow samples from 81 new diagnosed pediatric patients with AML, the cytogenetics and immunophenotypes of bone marrow cells were routinely examined. The results indicated that the FLT3-ITDs were detected in 8 out of 81 pediatric patients (9.88%) and all mutations detected were hybrid, while less frequently this mutation was detected in adult patients. Although they were irrelevant with sex and immunophenotypes, the mutations seemed predominant in older pediatric patients. The leukocyte counts and bone marrow blast cell counts in pediatric patients with FLT3-ITD at diagnosis were higher than those in pediatric patients without FLT3-ITD (p = 0.001 and p = 0.041 respectively), but the normal chromosomes were found in most pediatric patients with FLT-ITD. The patients with FLT3-ITD had lower induction remission rate (only 25%), but the patients without FLT3-ITD had higher remission rate (76.1%). According results detected by RT-PCR, the mdr1 gene was found in 27 pediatric patients, but only 3 out of 8 pediatric patients with FLT3-ITD were detected to express both FLT3-ITD and mdr1, which suggests unrelation between FLT3-ITD occurrence and mdr1 expression. It is concluded that the FLT3-ITD is frequent mutation in pediatric patients with AML, the prognosis is worse and the induction remission rate is lower in these patients, but the FLT3-ITD not relates with the mdr1, which suggests that the common MDR modulators may be un effective for therapy of the patients with FLT3-ITD.
ATP Binding Cassette Transporter, Sub-Family B
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ATP-Binding Cassette, Sub-Family B, Member 1
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genetics
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Adolescent
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Child
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Child, Preschool
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Drug Resistance, Multiple
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genetics
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Drug Resistance, Neoplasm
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genetics
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Female
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Gene Duplication
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Humans
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Leukemia, Myeloid, Acute
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genetics
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Male
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Mutation
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Prognosis
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Tandem Repeat Sequences
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fms-Like Tyrosine Kinase 3
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genetics
8.siRNA silences mdr1 gene expression and reverses apoptosis resistance of K562/ADM cells line.
Hu-lai WEI ; Li-ping GAO ; Tao JING ; Huai-shun ZHAO ; Juan YI ; Jin SUN ; Jian HAN
Chinese Journal of Hematology 2007;28(6):388-390
<b>OBJECTIVEb>To explore the effect of small interfering RNA(siRNA) on silence of mdr1 gene and reversal of apoptosis resistance in multidrug-resistant (MDR) human leukemia K562/ADM cell.
<b>METHODSb>Human MDR leukemia cell line K562/ADM was used as the target cells. Two siRNAs (mdr1 siRNA-1 and mdr1 siRNA-2) targeted mdr1 gene were chemically synthesized and transfected into K562/ADM cells with liposome. Expression of mdr1 mRNA was determined by real-time PCR, P-glycoprotein (P-gp) expression and caspase-3 activity were measured with flow cytometry (FCM), and the cell apoptosis was observed by optical and electronic microscopy for morphology and Annexin V/PI staining.
<b>RESULTSb>The mdr1 siRNA-1 and mdr1 siRNA-2 could markedly down-regulate the expression of mdr1 gene in K562/ADM cells, the expression of mdr1 mRNA decreased by 91.2% and 82.0% , and the P-gp by 74.1% and 84.4%, respectively. The caspase-3 activity was markedly enhanced, and the active caspase-3 in K562/ADM cells increased by about 40% compared to liposome alone and non-silencing controls. the sensitivity of K562/ADM cells to adriamycin-induced apoptosis was significantly augmented, the apoptotic rate of the cells treated with siRNA plus adriamycin increased by about 60% compared to adriamycin alone.
<b>CONCLUSIONb>siRNAs silence the expression of mdr1/P-gp to overcome the P-gp-mediated apoptosis resistance in drug-resistant K562/ADM cells.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Apoptosis ; Caspase 3 ; metabolism ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Humans ; K562 Cells ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection
9.Study on effect of berbamine on multidrug resistance leukemia K562/Adr cells.
Qing-hua DONG ; Shu ZHENG ; Rong-zhen XU ; Qinghua LU ; Liming HE
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(9):820-822
<b>OBJECTIVEb>To study the effect and mechanism of berbamine on the apoptosis of multidrug resistant leukemia K562/Adr cells and in reversing the drug resistance.
<b>METHODSb>IC50 value of K562/Adr cell was determined with MTT method, cell apoptosis rate was analyzed by flow cytometry with Annexin V FITC-PI assay, with the peak and cell cycle detected by PI staining. At the same time, flow cytometry was also used in determining Caspase-3, P-GP protein expression and drug accumulating capacity in cells, and RT-PCR method was used to analyze the gene expression of mdr-1.
<b>RESULTSb>Berbamine could inhibit human leukemia K562/Adr cell growth in dose-dependent manner, it could also induce cell apoptosis, increase the protein expression of Caspase-3 and the drug excretion capacity of cells, reduce the mRNA and protein expression levels of mdr-1 gene.
<b>CONCLUSIONb>Berbamine could activate Caspase-3 to induce human leukemia K562/Adr cell apoptosis, and by reducing mdr-1 gene expression to reverse its multidrug resistance.
ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; genetics ; Alkaloids ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Benzylisoquinolines ; pharmacology ; Caspase 3 ; Caspases ; biosynthesis ; genetics ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Humans ; K562 Cells ; RNA, Messenger ; biosynthesis ; genetics
10.Effect of cryptotanshinone on imatinib sensitivity and P-glycoprotein expression of chronic myeloid leukemia cells.
Yu-qing GE ; Ru-bin CHENG ; Bo YANG ; Zhen HUANG ; Zhe CHEN
China Journal of Chinese Materia Medica 2015;40(12):2389-2395
Cryptotanshinone (CPT), a lipid soluble active compound in Salvia miltiorrhiza, has a significant inhibitory effect on multiple malignant tumors, e. g. chronic myeloid leukemia (CML) cells and can effectively enhance imatinib's chemotherapeutic effect. However, its functional molecular mechanism remained unclear. In this experiment, the authors conducted a systematic study on the effect of CPT on the imatinib sensitivity and P-glycoprotein (P-gp) expression in CML cells by using CML cells K562 and imatinib persister K562-R. The MTT assays were performed to determine CPT's impact on the inhibitory effect of imatinib. Annexin V-FITC/PI staining analysis was used to detect the changes in the cell apoptosis rate. The active changes in apoptosis regulatory proteins Caspase-3, Caspase-9 and PARP were determined by Western blot. After the cells were pretreated with the gradient concentration of CPT, the expression of P-gp was analyzed by Western blot and flow cytometry. The changes in intracellular concentrations of imatinib were determined by HPLC analysis. The results indicated that the pretreatment with CPT significantly increased the proliferation inhibiting and apoptosis inducing effects of imatinib on K562 and K562-R cells as well as the degradation product expression of pro-apoptotic proteins Caspase-3, Caspase-9 and PARP, with a significant difference with the control group (P < 0.01). However, CPT showed no impact on the P-gp expression in CML cells and the intracellular concentrations of imatinib. In summary, the findings suggested that CPT enhanced the sensitivity of CML cells to imatinib. Its mechanism is not dependent on the inhibition in P-gp expression and the increase in intracellular drug concentration.
ATP-Binding Cassette, Sub-Family B, Member 1
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genetics
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metabolism
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Antineoplastic Agents
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pharmacology
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Apoptosis
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drug effects
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Caspase 3
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genetics
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metabolism
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Caspase 9
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genetics
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metabolism
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Drug Resistance, Neoplasm
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Humans
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Imatinib Mesylate
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pharmacology
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K562 Cells
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Leukemia, Myelogenous, Chronic, BCR-ABL Positive
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drug therapy
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genetics
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metabolism
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physiopathology
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Phenanthrenes
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pharmacology