Clinical characteristics and genetic analysis of a pedigree with Stargardt disease caused by a novel mutation in ABCA4 gene / 中华检验医学杂志

Objective:To explore the clinical characteristics and genetics of a pedigree with Stargardt disease, and investigate the pathogenicity of ABCA4 (ATP binding cassette subfamily A member 4) gene mutations in Stargardt disease.Methods:The proband was admitted to the Second People′s Hospital of Jinan in May 2021 due to diminution of vision. The proband was diagnosed with Stargardt disease according to the clinical diagnostic criteria of Stargardt disease. Detailed ophthalmological examinations was also performed on family members of the proband. Genomic DNA were extracted from the proband and the family members, and the whole exon sequencing was performed to find pathogenic gene mutations. The hazard of mutations was analyzed by polyphen-2, SIFT and MutationTaster websites. Sanger sequencing was used to verify the mutations. Conserved analysis of homologous species and 3-dimensional (3D) molecular model of the protein were used to analyze the pathogenicity.Results:Ophthalmological examinations showed reduced binocular vision, macular atrophy and "bull′s eye sign" in the proband and there was no abnormal signs and symptoms among the family members. Through whole exon sequencing analysis and Sanger sequencing verification, the compound heterozygous mutations (c.215G>A and c.6563T>C) of ABCA4 gene were co-segregated with this disease in this family. SIFT, Polyphen-2 and MutationTaster predicted that these two mutations were pathogenic. Conservative analysis and 3D molecular model of protein showed that mutations could cause changes in protein structure and affect protein function.Conclusion:The compound heterozygous mutations (C.215G>A and C.6563T>C) of ABCA4 gene are the pathogenic mutations of Stargardt disease in this pedigree.

Prenatal diagnosis of a fetus with harlequin ichthyosis caused by novel splice-site mutation in ABCA12 gene / 中华围产医学杂志

This article reported a rare case of harlequin ichthyosis which was indicated with multiple structural abnormalities by prenatal ultrasound and diagnosed by trio-based whole-exome sequencing (Trio-WES). Prenatal diagnosis was performed because the ultrasound at 24 +4 gestational weeks revealed the fetus presenting with eclabium, flattened nose, short mandible, small auricle and abnormal posture of the toes. Copy number variation sequencing (CNV-seq) showed no chromosome aneuploidy or pathogenic copy number variants over 100 kb in the fetal or parental samples. Trio-WES showed that the fetus carried two heterozygous mutations, c.2593-1G>A and c.7444C>T in ABCA12. Sanger sequencing confirmed that c.2593-1G>A, a previously unreported variant, was paternally inherited and c.7444C>T was maternally inherited. Both parents had normal phenotype. The fetus was finally diagnosed with harlequin ichthyosis. After prenatal counseling, the parents made an informed choice to terminate the pregnancy at 28 +4 gestational weeks. The stillborn fetus showed multiple malformations The variants in this case expand the spectrum of variants in ABCA12 gene.

Progression on Multifunctional Nanoparticles Interfering with Cell Membrane Transporters-associated Drug Resistance / 中国医学科学院学报

Multi-drug resistance(MDR)refers to the loss of sensitivity of tumor cells to traditional chemotherapeutics agents under the mediation of various mechanisms,resulting in the reduction of chemotherapy efficacy.Current studies suggest that a variety of factors,including cell membrane transporter-mediated efflux of anti-tumor drugs,special microenvironment in tumor tissue,DNA self-repair and anti-apoptotic process,and epithelial-mesenchymal cell transformation,may contribute to the formation of MDR.Cell membrane transporter-mediated drug efflux refers to an increase in the amount of anti-tumor drug pumped out of the cell through the up-regulation of the ATP-binding cassette transporter on tumor cell membrane,which reduces the concentration of the drug in the cell,thus forming MDR.An effective method to inhibit the efflux pump caused by overexpression of membrane transporters plays an important role in overcoming MDR.As a promising drug delivery system,multifunctional nanoparticles have demonstrated many advantages in antitumor therapy.Meanwhile,nanoparticles with tailored design are capable of overcoming MDR when combined with a variety of strategies.This paper described in detail the studies relevant to the use of multifunctional nano-sized drug delivery system combined with different strategies,such as co-delivery of agents,external responsiveness or target modification for intervention with efflux pump in order to reverse MDR.This paper provides reference for the development of nano-sized drug delivery system and the formulation of reversal strategy in the future.

The correlation between the expression of differential drug-resistant proteins and inflammatory cyto-kines in collagen-induced arthritis model / 中华风湿病学杂志

Objective We investigated the correlation between inflammatory cytokines and drugresi-stant proteins. Methods Fourteen DBA1 mice were successfully induced by collagen and Freund's adjuvant. According to the scores of synovial pathology, the collagen-induced arthritis (CIA) group was divided into mild, moderate-severe groups, another four mice were selected as controls. The mRNA expressions of P-glycoprotein (P-gp), breast cancer resistance protein (BCRP), multidrug resistance protein 1 (MRP1) in splenic lymphocyte cells were measured by reverse transcription-polymerase chain reaction (RT-PCR). The concentrations of inter-leukin (IL)-1β, IL-2, IL-6, IL-10, umor necrosis factor (TNF)-α, IL-17 in serum were detected by Cytometric Bead Array (CBA). The correlation between different inflammatory cytokines and these proteins were analyzed, then one of the proteins which were most related with cytokines by immunohisto-chemical (IHC) in the synovium was studied. Data were analyzed by the Mann-Whitney U test, Kruskal-Wallis H test and spearman analysis. Results ①Compared with normal controls, the levels of IL-6 and TNF-α in the serum of mild CIA group, moderate-severe CIA group were significantly increased {IL-6 controls: [7.75 (5.14, 9.17) pg/ml];mild CIA: [25.31 (15.15, 29.27) pg/ml]; modeate-severe CIA: [45.03 (38.87, 64.02) pg/ml]. TNF-α: controls: [22.81 (20.84, 28.17) pg/ml]; mild CIA: [45.00(32.76, 58.51) pg/ml]; modeate-severe CIA: [45.00(39.78, 8.95) pg/ml]}(Z=14.383, P<0.05; Z=8.375, P<0.05). Compared with the mild CIA group, the level of IL-6 in serum was significantly increased in the moderate-to-severe CIA group (P<0.05), but there was no signigicant difference in the TNF-α level (P>0.05). ② In the spleen lymphocytes, there was no significant difference in the mRNA expression level of P-gp and BCRP among the groups, but the mRNA expression level of MRP1 was significantly increased (Z=12.634, P<0.05). Compared with the mild CIA group, the MRP1 mRNA in the moderate-severe CIA group was higher, the difference was significant (Z=12.634, P<0.05). There was a correlation between mRNA expression of MRP1 and P-gp (r=0.635, P=0.015). ③ The mRNA expression of MRP1 was positively correlated with IL-6 level (r=0.711, P=0.004). ④ The expression of MRP1 in normal group, low-level IL-6 group and high- IL-6 level group were as follows: [1.08 (0.65, 1.30)], [1.32 (1.08, 1.49)], [2.07 (1.77, 2.22)] respectively.⑤Compared with the controls, the cytoplasm/membrane of the knee and ankle joint synovial tissue in the CIA group was yellowish-brown, which indicated that MRP1 expression was positive. Conclusion In the CIA arthritis model, synovial tissue lesion is not only related to inflammatory cytokines, but also related to MRP1 expression in the ATP-binding cassette (ABC) transport protein family, and it is proved that IL-6 is highly correlated to MRP1.

Effect of epigallocatechin-3 gallate on cholesterol efflux in foam cells and its mechanism / 中华老年心脑血管病杂志

Objective To study the effect of epigallocatechin-3 gallate (EGCG) on cholesterol efflux in foam cells and its mechanism.Methods THP-1 cells were induced to differentiate into macrophages which were then transformed to foam cells.Foam cells were divided into 0 μmol/L EGCG group,10 μmol/L EGCG group,30 μmol/L EGCG group,and 100 μmol/L EGCG group (1.5 × 106 in each group).Their cholesterol content was measured with a cholesterol test kit,apoA-I-mediated cholesterol efflux was assayed with a liquid scintillation counter,expression of ATP-binding cassette A1 (ABCA1) was detected by RT-PCR and Western blot respectively.Results The ABCA1 mRNA and protein expression levels and cholesterol efflux were significantly higher while the cholesterol content was significantly lower in 10 μmol/L EGCG group,30 μmol/L EGCG group,and 100 μmol/L EGCG group than in 0 μmol/L EGCG group (7.04％ ±0.21％,7.75％±0.17％ and 8.53％±0.18％ vs 3.37％±0.16％,P＜0.01;419.33±19.75 mg/g,352.58± 14.23 mg/g and 312.62±17.45 mg/g vs 520.51 ±20.62 mg/g,P＜0.01),and in 30 μmol/L EGCG group,100μmol/L EGCG group than in 10μmol/L EGCG group (P＜0.05).Conclusion EGCG increases cholesterol efflux and decreases cholesterol content in foam cells by upregulating the transcription and expression of ABCA1.

Expression changes and significance of ATP-binding cassette transporter B5 in pterygium / 中华实验眼科杂志

Background Limbal stem cells (LSCs) deficiency leads to many ocular surface diseases,such as pterygium and so on.ATP-binding cassette transporter B5 (ABCB5) is a recently discovered marker of LSCs.Understanding the expression changes of ABCB5 in pterygium tissue has an important clinical significance for pterygium.Objective This study was to investigate the expression changes of ABCB5 in pterygium tissue.Methods Thirty-seven pterygium tissue specimens were collected from primary pterygium patients who received pterygium surgery,and 22 normal conjunctival tissue specimens were obtained from the strabismus patients and retinal detachment patients during surgery in 474 Hospital of Chinese PLA from January 2015 to November 2015.Immunochemistry was employed to detect the cxpression and location of ABCB5 in the specimens,and the protein and mRNA expressions of ABCB5 were detected by Western blot and real-time fluorescence quantitative PCR,respectively.The results were compared between normal conjunctival tissues and pterygium tissue.Results ABCB5 protein was expressed in the cytoplasm and nuclei of stratified squamous epithelium of 22 normal counjunctival tissue specimens,especially in the basal layer cells,and showed obvious polarity in normal conjunctiva.In pterygium group,ABCB5 protein was positively expressed in 91.89％ specimens (34/37) and the expression was absent in 8.11％ specimens (3/37).The relative expression levels of ABCB5 protein were 0.90±0.31 and 0.59±0.41,and those of ABCB5 mRNA were 1.01±0.26 and 0.65±0.32 in the normal conjunctival tissues and pterygium tissue,respectively,showing significant differences between them (protein:t =-0.266,P =0.011;mRNA:t =-4.560,P =0.000).Conclusions Down-regulation of ABCB5 in pterygium indicates the decreasing and losing of LSCs,which may play an important role in the development of pterygium.ABCB5 may be a useful indicator for the prediction of development and recurrence of pterygium and has an important implication for treating evaluation,and it may also be a target for the management of pterygium.

The heredity of research in neonatal respiratory distress syndrome / 中国小儿急救医学

Neonatal respiratory distress syndrome(NRDS)is the most critical disease in neonatal pe-riod.Studies have proved that genetic factors play an important role in the pathogenesis of NRDS.More and more proteins and genes which are associated with NRDS are researched.This article mainly reviewed the re-search of surfactant protein,ATP-binding cassette transporters A3,mannose-binding lectin,thyroid transcrip-tion factor-1and NRDS.

Study on inhibitory action of proliferation of human lung carcinoma A549 cells with aqueous extract of Taxus chinensis.var combining Cisplatin and the effects on resistance genes / 中国医师杂志

Objective To investigate the inhibitory effect of aqueous extract of Taxus chinensis.vat (AETC) combining Cisplatin (DDP) on vitro cultured human lung carcinoma A549 cells,and the effects on resistance genes.Methods The A549 cells were divided into different concentrations of DDP groups,different concentrations of AETC groups,and blank group,and drug effect of 48 h with the method of cell counting kit-8 (CCK-8) and the effect on cell survival were detected.Based on the above results,then A549 cells were divided into DDP combining different concentrations of AETC groups,DDP group,blank control group,and drug effect of 48 h with the method of CCK-8 and the effect on cells survival were detected.The gene expressions of adenosine triphosphate (ATP)-binding cassette subfamily B member 1 trans-porter (ABCB1),ABCG2,and ABCC1 were examined by polymerase chain reaction (RT-PCR).Results Cisplatin 12 μg/ml (DDP),DDP + ATEC 400 μg/ml,DDP + ATEC 800 μg/ml,DDP + ATEC 1 200 μg/ml,DDP + ATEC 1 600 μg/ml,A549 cell inhibition rate of each group was 44.36％,69.61％,74.73％,80.10％,and 74.73％,respectively;Different concentrations of AETC combining DDP could decrease the resistance related gene ABCC1,ABCB1 expressions,and correlated to the dose.AETC combining DDP showed no effects on ABCG2 gene expression.Conclusions AETC combining DDP could inhibit the growth of A549 cells,and decrease the resistance-related gene ABCC1,ABCB1 expressions.

Gene expression profile of ABC transporters and cytotoxic effect of ibuprofen and acetaminophen in an epithelial ovarian cancer cell line in vitro / Perfil da expressão gênica dos transportadores ABC e efeito citotóxico do ibuprofeno e acetaminofen em uma linhagem celular de câncer epitelial de ovário in vitro

PURPOSES: To determine the basic expression of ABC transporters in an epithelial ovarian cancer cell line, and to investigate whether low concentrations of acetaminophen and ibuprofen inhibited the growth of this cell line in vitro. METHODS: TOV-21 G cells were exposed to different concentrations of acetaminophen (1.5 to 15 μg/mL) and ibuprofen (2.0 to 20 μg/mL) for 24 to 48 hours. The cellular growth was assessed using a cell viability assay. Cellular morphology was determined by fluorescence microscopy. The gene expression profile of ABC transporters was determined by assessing a panel including 42 genes of the ABC transporter superfamily. RESULTS: We observed a significant decrease in TOV-21 G cell growth after exposure to 15 μg/mL of acetaminophen for 24 (p=0.02) and 48 hours (p=0.01), or to 20 μg/mL of ibuprofen for 48 hours (p=0.04). Assessing the morphology of TOV-21 G cells did not reveal evidence of extensive apoptosis. TOV-21 G cells had a reduced expression of the genes ABCA1, ABCC3, ABCC4, ABCD3, ABCD4 and ABCE1 within the ABC transporter superfamily. CONCLUSIONS: This study provides in vitro evidence of inhibitory effects of growth in therapeutic concentrations of acetaminophen and ibuprofen on TOV-21 G cells. Additionally, TOV-21 G cells presented a reduced expression of the ABCA1, ABCC3, ABCC4, ABCD3, ABCD4 and ABCE1 transporters. .

OBJETIVOS: Determinar a expressão básica dos transportadores ABC em uma linhagem celular do câncer epitelial de ovário, e investigar se o acetaminofen e o ibuprofeno em baixas concentrações são capazes de inibir o crescimento desta linhagem celular in vitro. MÉTODOS: A linhagem celular TOV-21 G foi exposta a diferentes concentrações de acetaminofen (1,5 a 15 µg/mL) e ibuprofeno (2,0 a 20 µg/mL), de 24 a 48 horas. O crescimento celular foi avaliado utilizando-se um ensaio de viabilidade celular. A morfologia celular foi determinada por meio da microscopia de fluorescência. O perfil de expressão gênica foi estabelecido por um painel de 42 genes da superfamília de transportadores ABC. RESULTADOS: Observou-se um decréscimo significativo no crescimento das células TOV-21 G expostas a 15 µg/mL de acetaminofen durante 24 (p=0,02) e 48 horas (p=0,01), ou a 20 µg/mL de ibuprofeno por 48 horas (p=0,04). Ao avaliar a morfologia das células cultivadas, não foi observada evidência de apoptose extensiva. A linhagem de células estudada subexpressa os genes de ABCA1, ABCC3, ABCC4, ABCD3, ABCD4 e ABCE1 na superfamília de transportadores ABC. CONCLUSÕES: Este estudo fornece evidências in vitro referentes aos efeitos inibidores do crescimento de concentrações terapêuticas do acetaminofen e ibuprofeno na linhagem celular testada. Além disso, as células TOV-21 G apresentaram uma expressão reduzida de genes dos transportadores ABCA1, ABCC3, ABCC4, ABCD3, ABCD4 e ABCE1. .

Identification and characterization of stem cells in an ovarian cancer cell line and examination their drug resistance / 中华妇产科杂志

Objective To isolate side population (SP) cells from an established ovarian cancer (OC)cell line,characterize these cells,and examine their drug resistance. Methods SP and non-SP (NSP) cells were isolated by fluorescence-activated cell sorting (FACS),and cultured in differential conditions,then detected their SP ratio to compare their capability of differentiation and self-renewal. Moreover,SP and NSP cell tumorigenesis was examined by subcutaneous and intraperitoneal injection of these cells to nonobes ediabetic(NOD)-severe combined immundeficient(SCID)mice. Drug resistance to cisplatin was examined by cell counting kit-8 (CCK-8).Results SP cells could be isolated stablly and insistently. There was(4.81 ± 0.43)%of SP cells in the established OC cell line and(4.89 ± 0.33)%of SP cells after cultured the isolated SP cells in differentiation condition,and there was no significant different between these two quantities (P>0.05). However,after cultured the NSP cells,there was only (0.10 ± 0.03)%of SP cells which was significantly lower than that contained in the OC cell line(P<0.01). In the tumorigenesis assay 1.0 × 103 SP cells were injected subcutaneously and formed the xenografted tumors in 6 weeks(3/3),and 1.0×104 NSP cells were injected subcutaneously and did not form xenografted tumors in 12 weeks(0). The tumorigenic capability of SP cells was higher than that of NSP cells(P<0.01). Both the original and the xenografted tumors were low differentiated serous cystadenocarcinomas and expressed the ovarian serous cystadenocarcinomas CA125 marker after stained by HE and immunohistochemistry. Simultaneously,the SP cells were also capable to form tumors as shown by intraperitoneal injection. In the drug resistance assay shown that the 50% inhibitory concentration (IC50) of the SP and NSP cells were respectively(2.33 ± 0.14)μg/ml and(1.60 ± 0.04)μg/ml(P<0.05). After treated the unsorted OC cells with cisplatin,the quantity of SP cells increased to(40.10 ± 4.22)%and there was significant difference,when compared to the untreated cells which was(4.81±0.43)%(P<0.01). The SP cells survival rate was(58.7± 3.3)%when treated with cisplatin at its IC50 dose,and the rate decreased to(7.2±1.3)%(P<0.01)when verapamil was present. Conclusions The SP cells could be isolated from the established OC cell line. They had the capacities of self-renewal,differentiation,and tumorigenesis,and the new tumor demonstrated the original tumor′s phenotype. The SP cells also had stem cells′ biological characteristics and is resistant to cisplatin.

Effect of lipopolysaccharide on expression of ATP-binding cassette transporter A1 in alveolar macrophage cells of rats / 中华麻醉学杂志

Objective To evaluate the effect of lipopolysaccharide (LPS) on the expression of ATP-binding cassette transporter A1 (ABCA1) in alveolar macrophage cells of rats.Methods The alveolar macrophage cells of rats NR8383 were seeded in 6-well plates at a density of 1 × 106 cells/ml (2 ml/well) and randomly divided into 6 groups:control group (group C,n =24),0.2 μg/L LPS group (group L0.2,n =12),2.0 μg/L LPS group (group L2.0,n =12),20.0 μg/L LPS group (group L20.0,n =60),200.0 pg/L LPS group (group L200.0,n =12),and 20.0 μg/L LPS + ABCA1 siRNA group (group L20.0 + siRNA,n =12).The cells were routinely cultured in group C.LPS with the final concentrations of 0.2,2.0,20.0 and 200.0μg/L was added to the culture medium in L0.2,L2.0,L20.0 and I200.0 groups,respectively.In group L20.0 + siRNA,siRNA 50 nmol/L was added to the culture medium and 12 h later LPS 20.0 μg/L was added.In group C,6 wells were chosen for determination of ABCA1 mRNA and protein expression.At 24 h of incubation with LPS 0.2,2.0 and 200.0 μg/L,or at 2,6,12 and 24 h of incubation with LPS 20.0 μg/L,6 wells were chosen and the cell suspension was obtained for measurement of ABCA1 mRNA expression (by real-time fluorescent quantitative PCR),and ABCA1 expression (by flow cytometry).At 12 h of incubation with 20.0 μg/L LPS or with 20.0 μg/L LPS + 50 nmol/L siRNA,6 wells were chosen and the cell suspension was obtained for measurement of TLR4 mRNA expression (by real-time fluorescent quantitative PCR) and TLR4 expression (by flow cytometry).Results Compared with group C,the expression of ABCA1 mRNA and protein was significantly down-regulated in L0.2,L2.0,L20.0 and L200.0 groups,and the expression of ABCA1 mRNA and protein was up-regulated in L20.0 and L20.0 + siRNA groups.In L20.0 group,the expression of ABCA1 mRNA and protein was gradually down-regulated with the prolonging time of incubation with LPS.Compared with group L20.0,the expression of TLR4 mRNA and protein was significantly up-regulated in group L20.0 + siRNA.Conclusion LPS can down-regulate the expression of ABCA1 in alveolar macrophage cells of rats,however,ABCA1 can inhibit the synthesis of TLR4.

Effect of ABCE1 gene silencing by electroporation on proliferation and migration of human glioma cells line U87MG / 中国医师杂志

Objective To investigate the effect of ATP-binding cassette protein E 1 (ABCE1) gene silencing by electroporation on the survival,cell cycle and invasion of human glioma cells line U87MG.Methods The siRNA against ABCE1 was constructed and transfected into U87MG cells by electroporation.The expression of ABCE1 was detected by real time-polymerase chain reaction (RT-PCR) and Western blot.Flow cytometry was used to detect the cell cycle distribution and apoptosis.The effects of ABCE1 gene silencing by electroporation on proliferation,migration and invasion of U87MG cell line were evaluated by cell counting kit-8 (CCK-8) assay,wound closure assay,chemotactic migration,and cell invasive experiments,respectively.Results Compared to the control and blank groups,the mRNA and protein levels were significantly decreased in the experimental group when ABCE1 gene silencing by electroporation.The cell cycle was arrested at G0/G1 phase,and cell number in S phase was decreased in U87MG cell line (P ＜ 0.05).The cell growth inhibition ratio in the experimental group was significantly higher than that in the control and blank groups (P ＜0.01).Compared to the control and blank groups,the experimental group U87MG cell proliferation was inhibited significantly (P ＜ 0.05).Scratch healing experiments showed the experimental group migration ability was decreased significantly (P ＜ 0.05).Transwell chamber method showed the experimental group U87MG cell invasion ability was decreased significantly (P ＜ 0.05).Conclusions ABCE1 is involved in the progression of human glioma cells,and inhibiting the expression of ABCE1 by electroporation can decrease migration,invasion,and proliferation ability of tumor cells in vitro.

Polymorphism of ABCG5/ABCG8 gene (rs4299376) and lipid levels in Han Chinese with coronary heart disease / 第二军医大学学报

Objective: To explore the association between variant rs4299376 of ABCG5/ABCG8 gene with the risk of coronary heart disease (CHD) in Han Chinese and the association of related lipid levels with CHD. Methods: We collected blood samples from 290 CHD cases, 198 non-CHD controls and 331 healthy controls. The genomic DNA was acquired by the nucleic acid extraction automatic analyzer and the rs4299376 genotypes were analyzed by the Mass-ARRAY İPLEXR platform. Results: There were no significant differences in distribution of variant rs4299376 of ABCG5/ABCG8 gene among Han Chinese with CHD, non-CHD controls and healthy controls. There was no association between lipid levels and CHD in either total or male groups. While in female group, the triglyceride (TG) and total cholesterol (TC) were higher in CHD patients than in non-CHD controls (TG: 2. 23 ± 1. 05vs 1. 84 ± 1. 03, P = 0. 01; TC: 4. 7 9 ± 1. 17 vs 4. 36 ±1. 03, P = 0. 01). It was also found that, for those 60 years old and above, the CHD cases had a significantly lower high density lipoprotein (HDL) levll compared with non-CHD group (1. 09±0. 23 vs 1. 16±0. 25, P = 0. 03). Conclusion: There is no noticeable association between ABCG5/ ABCG8 rs4299376 polymorphism and the risk of CHD in Han Chinese. Female CHD patients have higher levels of TG and TC and when over 60 year old, CHD patients have a lower HDL level than non-CHD controls.

ATP-binding cassette transporter A1 R219K polymorphism and the risk of ischemic stroke in Chinese populations: a meta-analysis / 国际脑血管病杂志

Objective To investigate the correlation between ATP-binding cassette transporter A1 (ABCA1) R219K polymorphism and ischemic stroke in Chinese populations.Methods The case control studies of the correlation between Chinese ABCA1 R219K polymorphism and ischemic stroke published before May 2013 were collected using comprehensive literature search.The Stata 11.0 software was used to conduct metaanalysis.Odds ratio (OR) and its 95％ confidence interval (CI) was used to evaluate the strength of association between the gene polymorphism and ischemic stroke.Results A total of 10 studies met the criteria and were included in the analysis,including 1 619 patients in the patient group and 1907 in the control group.The selected literature had no obvious bias.Meta-analysis showed that the risk of ischemic stroke in patients carrying RK + KK genotype significantly decreased 8％ (OR 0.92,95％ CI 0.88-0.96; P =0.000)compared to those carrying RR genotype.The risk of ischemic stroke in patients carrying KK genotype significantly decreased 36％ compared to those carrying RR genotype (OR 0.64,95％ CI 0.44-0.94; P =0.02).The risk of ischemic stroke in patients carrying RK genotype significantly decreased 19％ compared to those carrying RR genotype (OR 0.81,95％ CI 0.69-0.95; P =0.009).The risk of ischemic stroke in patients carrying K allele significantly decreased 17％ compared to those carrying R allele (0R 0.83,95％ CI 0.69-0.99; P =0.036).Conclusions ABCA1 R219K polymorphism is associated with the susceptibility of ischemic stroke in Chinese.K allele may be a genetic protective factor for ischemic stroke in Chinese populations.

Expression and clinical significance of thymidylate synthase and adenosine triphosphate-binding cassette superfamily G member protein in advanced gastric carcinoma / 中华消化杂志

Objective To investigate the expressions of thymidylate synthase (TS) and adenosine triphosphate (ATP)-binding cassette superfamily G member 2 (ABCG2) in advanced gastric cancer (GC) and to explore their correlation with clinical pathological features.Methods A total of 80surgical specimens of advanced gastric cancer patients were collected.The expressions of TS and ABCG2 in gastric cancer tissues and adjacent normal gastric tissues were detected by immunohistochemical method.The expression of P-glycoprotein in gastric cancer tissues was also examined.The correlations between TS,ABCG2 and clinical pathological features and P-glycoprotein were analyzed.Chi-square test was performed for two groups comparison and Kruskal-Wallis H test were used for multi-groups comparison.Results The positive rates of both TS and ABCG2 in gastric cancer tissues [85.0％ (68/80) and 90.0％ (72/80)] were higher than those of adjacent normal gastric tissues [62.5 ％ (50/80) and 78.7 ％ (63/80)],the differences were statistically significant (x2 =11.466 and 16.463,P=0.009 and 0.001).There were close correlation between the expression of TS,ABCG2 and tumor TNM stage,differentiation status,invasion depth (TS:x2 =30.686,61.470 and 40.545 ; ABCG2:x2=48.192,63.150 and 47.512; all P＜0.01).The later the tumor staged,the worse the cells differentiated and the deeper the tumor invaded,the higher level they expressed.Both TS and ABCG2expressions in gastric cancer tissues were correlated with the expression level of P glycoprotein (x2 =43.977and 29.509,both P＜0.01).Conclusion TS and ABCG2 may be potential indexes to predict the degree of malignancy,progression,drug resistance and prognosis in gastric cancer.

Study of the invasiveness and tumour formation of Bcrp1 + HeLa cervical cancer cells / 中华妇产科杂志

Objective To make sure whether or not Bcrp1 is the marker of cervical cancer stem cells or not by studying the invasive ability and formation of tumors of Bcrp1 + phenotype HeLa cells.Methods The tumor cell migration and invasion assay were used by boyden chamber to identify the invasive ability of Bcrp1 + phenotype HeLa cells.The formation of tumors in vivo experiments were completed,in which the two groups of cells with different concentrations were inoculated in non obese diabetes-severe combined immunodeficiency disease ( NOD/SCID ) mice ( 1 × 104,1 × 105,1 × 106/ml ) and the differences of time,rate and volume in the formation of tumors between two groups were observed.Results ( 1 ) In the invasion assay,the amount of cells that invaded through the artificial basement membrane in Bcrp1 + group were 99 ± 14,which was significantly greater than those in Bcrp1- group ( 57 ± 13,P ＜ 0.05 ) ; the length of the Bcrp1 + group was ( 366 ± 52 ) μm,which was significantly greater than the Bcrp1 - group ( 301 ± 54) μm ( P ＜ 0.05 ).( 2 ) Following transplantation of 1 × 104 cells,only the Bcrp1 + cells formed tumors in NOD/SCID mice.When 1 × 105 or 1 × 106 cells were transplanted,the tumor incidence and the tumor mass were greater in the Bcrp1 + groups than those in the Bcrp1 - groups ( P ＜ 0.05 ).Conclusion Bcrp1 + HeLa cell have the greater capacity of invasive and the tumorigenicity,which may contain cancer stem cells.

Establishment of cisplatin-resistant human endometrial cancer cell line and the study of its resistant mechanisms / 中华妇产科杂志

Objective To establish the cisplatin(DDP)-resistant cell line from human endometrial cancer cell line Ishikawa and to investigate its resistant mechanism to DDP. Methods A resistant endometrial cancer cell line ISH/DDP was established by gradually increasing dose of cisplatin and high-dose stimulation. The resistant index was estimated by 3-(4,5-dimethylthiazol-zyl)-5-(3-carboxymethoxyphenyl-2-(4-sulfophenyl)-2H-tetrazolium,inner salt (MTS) assay. Cell growth curve, doubling time and cell cycle phase distribution were measured; drug-resistant protein of breast cancer resistance protein (BCRP),P-glycoprotein (P-gp) and glutathione-S-transferase-π (GST-π) were examined by immuocytochemistry. Results The DDP-resistant cell line ISH/DDP was established with the resistant index of 3. 77. The proliferation of ISH/DDP got slow, doubling time prolonged, which were 40. 1 hours, while it was 34. 1 hours in Ishikawa (P0. 05] and G2/M phase [(11.9 ±0.7)% and (5. 7 ±2. 4)% ,P 0. 05) ,respectively. The score of the expression of GST-π in ISH/DDP and Ishikawa were 15. 2 ± 1. 9 and 14. 9 ± 1.1 (P > 0. 05) . Conclusion ISH/DDP cell line showed a typical resistant phenotype and biological characteristics, which may be accounted for high BCRP expression.

Association of drug efflux pump gene expression with Mycobacterium tuberculosis drug resistance / 中华检验医学杂志

Objectives To explore the associations between drug efflux pump gene expression and phenotypic drug resistance as well as gene mutation patterns related to drug resistance of Mycobacterium tuberculosis.Methods Forty-five Mycobacterium tuberculosis isolates resistant to one or more of drugs including isoniazid, rifampicin, streptomycin and ethambutol, and 26 isolates all sensitive to the above four drugs from Tianjin Tuberculosis Control Institute in 2007 were involved in this study. Direct sequencing was applied to detect the mutations in the corresponding resistance genes(isoniazid:katG, inhA, oxyR-ahpC, ndh, rifampicin:rpoB, streptomycin:rpsL, rrs, and ethambutol:embB, embC and embA). After RNA extration and reverse transcription, real-time PCR was conducted to assess the expressions of putative drug efflux pump genes Rv1410c, Rv2136c, Rv0783c and Rv2136c, and Students' t test and ANOVA analysis were used to analyze the expression differences in Mycobacterium tuberculosis with different phenotypic drug resistance and drug resistance related gene mutation patterns.Results Compared to pan-sensitive isolates[(5.67±3.29)×10-5], Rv1410c showed higher expression in streptomycin[(8.48±6.33)×10-5, t'=2.18, P<0.05], isoniazid[(8.43±6.38)×10-5, t'=2.20, P<0.05], rifampicin[(9.59±7.27)×10-5, t'=2.29, P<0.05], multi-drug[(10.37±7.86)×10-5, t'=2.34, P<0.05] resistant isolates, and in isoniazid + streptomycin resistant isolates[(9.39±6.81)×10-5, t'=2.43, P<0.05];Rv2136c showed higher expression in isoniazid resistant[(3.51±2.43)×10-5, t'=2.03, P<0.05], multidrug-resistant isolates[(4.21±2.94)×10-5, t'=2.22, P<0.05] and resistant to isoniazid+streptomycin[(3.81±2.46)×10-5, t'=2.28, P<0.05] isolates . The expression of Rv0783c in rifampicin resistant isolates with rpoB 531 mutations [(5.41±3.03)×10-6] was higher than those with wild type of rpoB 531[(2.29±1.62)×10-6, t=2.81, P<0.05].Conclusions The expression of Rv1410c and Rv2136c are associated with mutiple-drug resistance of Mycobacterium tuberculosis.The expression of Rv0783c in rifampicin resistant isolates is associated with mutation in rpoB 531.

Lipid raft involved in drug resistance: Relationship between multidrug resistance ATP-binding cassette transporters and lipid raft / 第二军医大学学报

Lipid rafts have been implicated in many cellular functions, including protein and lipid transport and signal transduction. Recently ATP-binding cassette (ABC) transporters, which are associated with multidrug resistance, have been found in lipid rafts; therefore they might be related to drug resistance. Here we introduce the relationship between the localization and functions of three multi-drug related ABC transporters, including two relevant to multidrug resistance in tumor cells(Pgp/ABCB1 and MRP1/ABCC1) and one relevant to multidrug resistance in Candida albicans (Cdrlp). We also discuss the influence of sphingolipids and cholesterol, two major components of lipid rafts, on the localization and function of the above three ABC transporters.

Effects of apolipoproteinA1 on reverse cholesterol transport and expression of ATP-binding cassette transporter A1 in human acute monocytic leukemia cell line macrophage-derived foam cells / 中华老年医学杂志

Objective To investigate the effects of apolipoproteinA1 (apoA1) on levels of cholesterol, cholesteryl ester (CE), and expression of ATP-bindiag cassette transporter A1 (ABCA1) in human acute monocytie leukemia cell line (THP-1) macrophage-derived foam cells.Methods The cultured THP-1 cells were induced into foam cells by exposing first to phorbol myristate acetate (PMA, 50 ng/ml) for 48 h, and then to oxidized-low density lipoprotein (ox-LDL, 50μg/ml) for 48 h.Under treatment of apoA1 in different doses (5, 10, 15 and 20 μg/ml) and one simple dose (10 μg/ml) for different time (6, 12 and 24 h), THP-1 macrophage-derived foam cells were incubated to observe the expression of cholesterol and ABCA1.The concentrations of cellular total cholesterol (TC), free cholesterol (FC) and CE were determined by oxidization enzymatic methods.Oil red O dyeing experiment was used to show the cellular lipid droplets in the cells.The expression of ABCA1 was tested by immunofluorescence method.Reverse transcription-polymerase chain reaction was applied to investigate mRNA expression of ABCA1.Results The THP-1 cells turned into typical foam cells after treated with PMA (50 ng/ml) for 48 h, and ox-LDL (50 μg/ml) for 48 h.apoA1 could lower the levels of TC, FC and CE in THP-1 macrophage-derived foam cells in a dose-dependent and a time-dependant manner, apoA1 could increase the expression of ABCA1 protein in THP-1maerophage-derived foam cells without up-regulation of mRNA.Antibody of ABCA1 could up- regulate the expression of ABCA1.Conclusions apoA1 may decrease the levels of cholesterols in THP-1 macrophage-derived foam cells, by promoting the expression of ABCA1 and the reverse cholesterol transport of high density lipoprotein.