目的探究利多卡因体外下调ATP结合转运蛋白G家族成员2(ATP binding cassette transporter G family member 2,ABCG2)抑制人乳腺癌细胞的活性及顺铂耐药性的可能机制。方法选取MDA-MB-231、MCF-7细胞,使用CCK-8、EdU、TUNEL法染色验证利...目的探究利多卡因体外下调ATP结合转运蛋白G家族成员2(ATP binding cassette transporter G family member 2,ABCG2)抑制人乳腺癌细胞的活性及顺铂耐药性的可能机制。方法选取MDA-MB-231、MCF-7细胞,使用CCK-8、EdU、TUNEL法染色验证利多卡因对细胞增殖活性、顺铂干预增敏作用,RT-PCR和Western印迹法检测利多卡因对ABCG2以及凋亡蛋白、耐药蛋白表达及PI3K/Akt蛋白磷酸化的影响;CCK-8法、TUNEL法验证过表达ABCG2对利多卡因联合顺铂干预乳腺癌细胞系增殖活性、凋亡率。结果0.5 mmol/L干预时乳细胞存活率及EdU阳性细胞数显著降低(P<0.05);与对照组相比,利多卡因组和顺铂组细胞存活率、Bcl-2表达显著降低,细胞凋亡率、Cleaved-PARP、Cleaved-caspase-3、Bax表达量显著升高,联合处理后变化更显著(P<0.05);利多卡因处理后,癌细胞系中ABCG2 mRNA及蛋白表达水平降低(P<0.05);与对照组比较,顺铂+利多卡因组ABCG2、P-gp、MRP1、MRP2蛋白表达显著降低(P<0.05);与顺铂组比较,利多卡因组细胞存活率、p-PI3K/PI3K、p-AKT/AKT表达显著降低(P<0.05),细胞凋亡率显著升高(P<0.05);与空质粒组比较,ABCG2过表达组细胞存活率、p-PI3K/PI3K、p-AKT/AKT表达显著升高(P<0.05),细胞凋亡率显著降低(P<0.05)。结论利多卡因可能通过抑制ABCG2表达及PI3K/AKT信号通路的激活抑制乳腺癌细胞增殖活性及提高对顺铂敏感性。展开更多
目的:研究微小RNA(microRNA,miR)-218-5p靶向调控三磷酸腺苷结合盒超家族G家族第2个成员(ATP-binding cassette sub-family G member 2,ABCG2)基因对Tca-8113细胞的增殖、迁移和侵袭的影响,并探讨其机制。方法:运用实时荧光定量聚合酶...目的:研究微小RNA(microRNA,miR)-218-5p靶向调控三磷酸腺苷结合盒超家族G家族第2个成员(ATP-binding cassette sub-family G member 2,ABCG2)基因对Tca-8113细胞的增殖、迁移和侵袭的影响,并探讨其机制。方法:运用实时荧光定量聚合酶链反应(quantitative Real-time PCR,qRT-PCR)与Western blot检测Tca-8113细胞和NOK细胞中miR-218-5p和ABCG2的表达。通过双荧光素酶报告基因实验检测miR-218-5p和ABCG2靶向关系。用脂质体法将miR-218-5p mimic、ABCG2小干扰RNA(interfering RNA,siRNA)、miR-218-5p mimic+pcDNA-ABCG2分别转染至Tca-8113细胞中,噻唑蓝(methyl thiazol tetrazolium,MTT)法、Transwell法检测Tca-8113细胞的增殖、迁移和侵袭。结果:与NOK细胞比较,Tca-8113细胞miR-218-5p呈显著低表达,ABCG2呈显著高表达(P<0.05)。过表达miR-218-5p、抑制ABCG2均可抑制Tca-8113细胞的增殖和迁移(P<0.05)。双荧光素酶报告基因实验证实miR-218-5p与ABCG2的靶向结合关系。过表达ABCG2可逆转上调miR-218-5p对Tca-8113细胞增殖、迁移和侵袭的抑制作用。结论:上调miR-218-5p的表达可靶向调控ABCG2,抑制Tca-8113细胞的增殖、迁移和侵袭,将可为口腔鳞状细胞癌的治疗提供新的靶点。展开更多
[目的]旨在检测三磷酸腺苷结合转运蛋白G超家族成员2(ABCG2)基因Y367C位点此在三河牛群体中的多态性,并分析了其与产奶性状之间的关系。[方法]利用Sequen-om Mass ARRAY Genotype分析技术对该位点进行检测,通过SAS软件(9.1)对6个产奶性...[目的]旨在检测三磷酸腺苷结合转运蛋白G超家族成员2(ABCG2)基因Y367C位点此在三河牛群体中的多态性,并分析了其与产奶性状之间的关系。[方法]利用Sequen-om Mass ARRAY Genotype分析技术对该位点进行检测,通过SAS软件(9.1)对6个产奶性状与该位点的关联程度进行了统计分析。[结果]表明,Y367C位点在三河牛中均处于Hardy-Weinberg平衡状态(P<0.05);ABCG2基因Y367C位点的YY基因型个体体细胞评分最低,CC个体产奶量、乳脂量、乳蛋白量最高;Y等位基因降低体细胞评分,提高乳蛋白率;而C等位基因则有助于提高产奶量、乳脂率、乳脂量和乳蛋白量,但三种基因型之间差异不显著(P<0.05)。[结论]该位点可以为三河牛的分子育种提供参考。展开更多
背景与目的:三磷酸腺苷结合转运蛋白G超家族成员2(adenosine triphosphate-binding cassette superfamily G member 2,ABCG2)在多种肿瘤细胞中表达,能通过外排抗癌药物参与肿瘤耐药。本研究的目的旨在探讨人胶质瘤细胞对焦脱镁叶绿酸甲...背景与目的:三磷酸腺苷结合转运蛋白G超家族成员2(adenosine triphosphate-binding cassette superfamily G member 2,ABCG2)在多种肿瘤细胞中表达,能通过外排抗癌药物参与肿瘤耐药。本研究的目的旨在探讨人胶质瘤细胞对焦脱镁叶绿酸甲酯(pyropheophorbide-a methyl ester,MPPa)介导的光动力疗法(photodynamic therapy,PDT)杀伤效应的敏感性及其与ABCG2的关系。方法:选取处于对数生长期的胶质瘤细胞株U87、A172,分别经MPPa-PDT或MPPa-PDT+烟曲霉毒素C(fumitremorgin C,FTC)处理后,采用CCK-8法检测细胞活性;采用蛋白[质]印迹法(Western blot)检测细胞内ABCG2的表达;流式细胞技术法检测未光照前各组细胞内MPPa的含量;AnnexinⅤ-FITC/PI双染流式细胞术检测细胞凋亡率;DCFH-DA染色观察细胞内活性氧(reactive oxygen species,ROS)的产生。结果:MPPa-PDT能抑制A172、U87细胞的活性,且呈一定的光能量依赖性,A172达到半数致死量所需光能量密度为U87的8倍;A172较U87细胞对MPPa-PDT不敏感;A172细胞内高表达的ABCG2影响MPPa在细胞内的聚集;抑制ABCG2后,不仅可以增强MPPa-PDT对A172细胞的杀伤作用,同时可增加MPPa-PDT触发产生的ROS的量及细胞对MPPa的摄取。结论:人胶质瘤细胞株A172对MPPa-PDT相对不敏感,并且产生这种现象的机制可能是ABCG2外排MPPa,减少MPPa的细胞内聚集,进而减弱光敏剂活化后对肿瘤细胞的杀伤作用。展开更多
目的肿瘤高表达多药耐药相关转运蛋白三磷酸腺苷结合转运蛋白G家族成员2(ATP-binding cassette super family G member 2,ABCG2)是造成耐药及化疗失败的主要原因。文中旨在探讨环磷酰胺体内干预后肾上腺皮质癌细胞中ABCG2表达的意义。方...目的肿瘤高表达多药耐药相关转运蛋白三磷酸腺苷结合转运蛋白G家族成员2(ATP-binding cassette super family G member 2,ABCG2)是造成耐药及化疗失败的主要原因。文中旨在探讨环磷酰胺体内干预后肾上腺皮质癌细胞中ABCG2表达的意义。方法 BALB/C-nu裸鼠共10只,雌雄不拘,随机数表法分为对照组和环磷酰胺组,每组5只。将SW-13细胞注射入裸鼠皮下建立裸鼠皮下移植瘤模型,环磷酰胺组予腹腔注射环磷酰胺,对照组予注射等量等渗盐水,并对获得的2组瘤体组织进行原代培养。用免疫组化法(阳性肿瘤细胞百分比<10%者为阴性,≥10%为阳性)和流式细胞法分别检测瘤体组织及原代细胞株ABCG2的表达。结果流式细胞法检测ABCG2在环磷酰胺组细胞中表达率[(97.89±1.36)%]高于对照组细胞[(81.88±8.31)%],差异有统计学意义(P<0.05),免疫组化结果提示环磷酰胺组瘤体组织中ABCG2表达阳性率[(69.1±1.83)%]高于对照组[(53.4±1.65)%],差异有统计学意义(P<0.05)。结论多药耐药基因ABCG2参与肾上腺皮质癌耐药,可能为肾上腺皮质癌干预靶点。展开更多
Mitofusin2 （Mfn2） plays a pivotal role in the proliferation and apoptosis of vascular smooth muscle cells （VSMCs）. The purpose of this study was to investigate the effects of Mfn2 on the traffick- ing of intracell...Mitofusin2 （Mfn2） plays a pivotal role in the proliferation and apoptosis of vascular smooth muscle cells （VSMCs）. The purpose of this study was to investigate the effects of Mfn2 on the traffick- ing of intracellular cholesterol in the foam ceils derived from rat VSMCs （rVSMCs） and also to investigate the effects of Mfn2 on the expression of adenosine triphosphate-binding cassette subfamily A member 1 （ABCA1）, adenosine triphosphate-binding cassette subfamily G member 1 （ABCG1） and peroxisome proliferator-activated receptor gamma （PPARy）. The rVSMCs were co-cultured with oxi- dized low density lipoprotein （LDL, 80 ~tg/mL） to produce foam cells and cholesterol accumulation in cells. Before oxidized LDL treatment, different titers （20, 40 and 60 pfu/cell） of recombinant adenovirus containing Mfn2 gene （Adv-Mfn2） were added into the culture medium for 24 h to transfect the Mfn2 gene into the rVSMCs. Then the cells were harvested for analyses. The protein expression of Mfn2 was significantly higher in Adv-Mfn2-transfected group than in untransfected group （P〈0.05）, and the ex- pression levels significantly increased when the titer of Adv-Mfn2 increased （P〈0.05）. At 24 or 48 h af- ter oxidized LDL treatment, rVSMCs became irregular and their nuclei became larger, and their plasma abounded with red lipid droplets. However, the number of red lipid droplets was significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group. At 48 h after oxidized LDL treatment, the intracellular cholesterol in rVSMCs was significantly increased （P〈0.05）, but it was sig- nificantly decreased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）, and it also significantly decreased when the titer of Adv-Mfn2 increased （P〈0.05）. The mRNA and pro- tein expression levels of ABCA1 and ABCG1 were significantly increased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）. Though the mRNA and protein expression levels of PPARy was not significantly increased （P〉0.05）, the phosporylation levels of PPARy were signifi- cantly decreased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）. These results suggest that the transfection of Adv-Mfn2 can significantly reduce intracellular cholesterol in oxidized LDL-induced rVSMCs possibly by decreasing PPAR＇/phosporylation and then increasing pro- tein expression levels of ABCAI and ABCG1, which may be helpful to suppress the formation of foam cells.展开更多
目的探讨S期激酶相关蛋白2(S-phase kinase interacting protein 2,Skp2)与三磷酸腺苷结合蛋白G亚族成员2(ATP-binding cassette sub-family G member 2,ABCG2)在鼻咽癌组织中的表达水平,及其与临床病理因素的关系。方法收集2013-01-12-...目的探讨S期激酶相关蛋白2(S-phase kinase interacting protein 2,Skp2)与三磷酸腺苷结合蛋白G亚族成员2(ATP-binding cassette sub-family G member 2,ABCG2)在鼻咽癌组织中的表达水平,及其与临床病理因素的关系。方法收集2013-01-12-2014-03-30,韶关市粤北人民医院耳鼻喉科行病理活检的63例鼻咽癌组织及20例正常鼻黏膜组织,采用免疫组织化学染色检测Skp2和ABCG2蛋白的表达,统计分析Skp2和ABCG2蛋白表达间及与肿瘤临床病理因素间的相关性。结果在鼻咽癌组织中,Skp2蛋白阳性表达率为74.6%(47/63),ABCG2为61.9%(39/63),均较正常鼻黏膜组织的20.0%(4/20)和15.0%(3/20)显著升高,P值均<0.001。在鼻咽癌组织中,Skp2和ABCG2蛋白表达呈显著正相关关系,r=0.751,P<0.001。高表达Skp2和ABCG2蛋白与肿瘤淋巴结转移(P值分别为0.003和<0.001)及高TNM分期(P值分别为0.003和0.013)呈显著正相关。结论 Skp2和ABCG2蛋白在鼻咽癌组织中表达上调,并与肿瘤恶性临床病理特征有关。Skp2和ABCG2蛋白可能成为鼻咽癌早期诊断的重要标志物及生物靶向治疗的有效靶点之一。展开更多
文摘目的探究利多卡因体外下调ATP结合转运蛋白G家族成员2(ATP binding cassette transporter G family member 2,ABCG2)抑制人乳腺癌细胞的活性及顺铂耐药性的可能机制。方法选取MDA-MB-231、MCF-7细胞,使用CCK-8、EdU、TUNEL法染色验证利多卡因对细胞增殖活性、顺铂干预增敏作用,RT-PCR和Western印迹法检测利多卡因对ABCG2以及凋亡蛋白、耐药蛋白表达及PI3K/Akt蛋白磷酸化的影响;CCK-8法、TUNEL法验证过表达ABCG2对利多卡因联合顺铂干预乳腺癌细胞系增殖活性、凋亡率。结果0.5 mmol/L干预时乳细胞存活率及EdU阳性细胞数显著降低(P<0.05);与对照组相比,利多卡因组和顺铂组细胞存活率、Bcl-2表达显著降低,细胞凋亡率、Cleaved-PARP、Cleaved-caspase-3、Bax表达量显著升高,联合处理后变化更显著(P<0.05);利多卡因处理后,癌细胞系中ABCG2 mRNA及蛋白表达水平降低(P<0.05);与对照组比较,顺铂+利多卡因组ABCG2、P-gp、MRP1、MRP2蛋白表达显著降低(P<0.05);与顺铂组比较,利多卡因组细胞存活率、p-PI3K/PI3K、p-AKT/AKT表达显著降低(P<0.05),细胞凋亡率显著升高(P<0.05);与空质粒组比较,ABCG2过表达组细胞存活率、p-PI3K/PI3K、p-AKT/AKT表达显著升高(P<0.05),细胞凋亡率显著降低(P<0.05)。结论利多卡因可能通过抑制ABCG2表达及PI3K/AKT信号通路的激活抑制乳腺癌细胞增殖活性及提高对顺铂敏感性。
文摘[目的]旨在检测三磷酸腺苷结合转运蛋白G超家族成员2(ABCG2)基因Y367C位点此在三河牛群体中的多态性,并分析了其与产奶性状之间的关系。[方法]利用Sequen-om Mass ARRAY Genotype分析技术对该位点进行检测,通过SAS软件(9.1)对6个产奶性状与该位点的关联程度进行了统计分析。[结果]表明,Y367C位点在三河牛中均处于Hardy-Weinberg平衡状态(P<0.05);ABCG2基因Y367C位点的YY基因型个体体细胞评分最低,CC个体产奶量、乳脂量、乳蛋白量最高;Y等位基因降低体细胞评分,提高乳蛋白率;而C等位基因则有助于提高产奶量、乳脂率、乳脂量和乳蛋白量,但三种基因型之间差异不显著(P<0.05)。[结论]该位点可以为三河牛的分子育种提供参考。
文摘目的肿瘤高表达多药耐药相关转运蛋白三磷酸腺苷结合转运蛋白G家族成员2(ATP-binding cassette super family G member 2,ABCG2)是造成耐药及化疗失败的主要原因。文中旨在探讨环磷酰胺体内干预后肾上腺皮质癌细胞中ABCG2表达的意义。方法 BALB/C-nu裸鼠共10只,雌雄不拘,随机数表法分为对照组和环磷酰胺组,每组5只。将SW-13细胞注射入裸鼠皮下建立裸鼠皮下移植瘤模型,环磷酰胺组予腹腔注射环磷酰胺,对照组予注射等量等渗盐水,并对获得的2组瘤体组织进行原代培养。用免疫组化法(阳性肿瘤细胞百分比<10%者为阴性,≥10%为阳性)和流式细胞法分别检测瘤体组织及原代细胞株ABCG2的表达。结果流式细胞法检测ABCG2在环磷酰胺组细胞中表达率[(97.89±1.36)%]高于对照组细胞[(81.88±8.31)%],差异有统计学意义(P<0.05),免疫组化结果提示环磷酰胺组瘤体组织中ABCG2表达阳性率[(69.1±1.83)%]高于对照组[(53.4±1.65)%],差异有统计学意义(P<0.05)。结论多药耐药基因ABCG2参与肾上腺皮质癌耐药,可能为肾上腺皮质癌干预靶点。
基金supported by the National Natural Science Foundation of China(No.30971244)
文摘Mitofusin2 （Mfn2） plays a pivotal role in the proliferation and apoptosis of vascular smooth muscle cells （VSMCs）. The purpose of this study was to investigate the effects of Mfn2 on the traffick- ing of intracellular cholesterol in the foam ceils derived from rat VSMCs （rVSMCs） and also to investigate the effects of Mfn2 on the expression of adenosine triphosphate-binding cassette subfamily A member 1 （ABCA1）, adenosine triphosphate-binding cassette subfamily G member 1 （ABCG1） and peroxisome proliferator-activated receptor gamma （PPARy）. The rVSMCs were co-cultured with oxi- dized low density lipoprotein （LDL, 80 ~tg/mL） to produce foam cells and cholesterol accumulation in cells. Before oxidized LDL treatment, different titers （20, 40 and 60 pfu/cell） of recombinant adenovirus containing Mfn2 gene （Adv-Mfn2） were added into the culture medium for 24 h to transfect the Mfn2 gene into the rVSMCs. Then the cells were harvested for analyses. The protein expression of Mfn2 was significantly higher in Adv-Mfn2-transfected group than in untransfected group （P〈0.05）, and the ex- pression levels significantly increased when the titer of Adv-Mfn2 increased （P〈0.05）. At 24 or 48 h af- ter oxidized LDL treatment, rVSMCs became irregular and their nuclei became larger, and their plasma abounded with red lipid droplets. However, the number of red lipid droplets was significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group. At 48 h after oxidized LDL treatment, the intracellular cholesterol in rVSMCs was significantly increased （P〈0.05）, but it was sig- nificantly decreased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）, and it also significantly decreased when the titer of Adv-Mfn2 increased （P〈0.05）. The mRNA and pro- tein expression levels of ABCA1 and ABCG1 were significantly increased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）. Though the mRNA and protein expression levels of PPARy was not significantly increased （P〉0.05）, the phosporylation levels of PPARy were signifi- cantly decreased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）. These results suggest that the transfection of Adv-Mfn2 can significantly reduce intracellular cholesterol in oxidized LDL-induced rVSMCs possibly by decreasing PPAR＇/phosporylation and then increasing pro- tein expression levels of ABCAI and ABCG1, which may be helpful to suppress the formation of foam cells.
文摘目的 考察急性淋巴细胞白血病(ALL)患儿腺苷三磷酸结合盒亚家族C成员2(ABCC2) rs717620 G>A基因多态性对甲氨蝶呤(MTX)血清浓度和化疗毒性的影响。方法 收集ALL患儿的外周血,提取基因组DNA,用基质辅助激光解吸电离飞行时间质谱技术分析ABCC2 rs717620 G>A基因型,用荧光偏振免疫法测定MTX血清浓度,记录MTX化疗后的ALL复发和毒性发生情况,分析ABCC2 rs717620 G>A基因多态性与剂量校正的MTX浓度(C/D比值)、复发和化疗毒性的相关性。结果 研究共纳入患儿127例,rs717620 GG、GA和AA基因型的分布频率分别为82.68%、16.54%、0.78%;G和A等位基因的分布频率分别为90.94%和9.06%。GG基因型患儿的24 h中位C/D比值(11.94μmol·L^(-1) per g·m^(-2))低于GA和AA基因型患儿(12.64μmol·L^(-1) per g·m^(-2)),中位42 h C/D比值(0.08μmol·L^(-1) per g·m^(-2))高于GA和AA基因型患儿(0.07μmol·L^(-1) per g·m^(-2)),复发率(11.42%)低于GA和AA基因型患儿(18.18%),在统计学上差异均无统计学意义(均P>0.05)。GG基因型患儿的血液毒性发生率(40.95%)和电解质紊乱发生率(21.90%)显著高于GA和AA基因型患儿(分别为13.64%和0.00%,P<0.05),其余不良事件的发生率在统计学上差异均无统计学意义(均P>0.05)。结论 ABCC2 rs717620 GG基因型可能是ALL患儿发生血液毒性和电解质紊乱的危险因素。
文摘目的探讨S期激酶相关蛋白2(S-phase kinase interacting protein 2,Skp2)与三磷酸腺苷结合蛋白G亚族成员2(ATP-binding cassette sub-family G member 2,ABCG2)在鼻咽癌组织中的表达水平,及其与临床病理因素的关系。方法收集2013-01-12-2014-03-30,韶关市粤北人民医院耳鼻喉科行病理活检的63例鼻咽癌组织及20例正常鼻黏膜组织,采用免疫组织化学染色检测Skp2和ABCG2蛋白的表达,统计分析Skp2和ABCG2蛋白表达间及与肿瘤临床病理因素间的相关性。结果在鼻咽癌组织中,Skp2蛋白阳性表达率为74.6%(47/63),ABCG2为61.9%(39/63),均较正常鼻黏膜组织的20.0%(4/20)和15.0%(3/20)显著升高,P值均<0.001。在鼻咽癌组织中,Skp2和ABCG2蛋白表达呈显著正相关关系,r=0.751,P<0.001。高表达Skp2和ABCG2蛋白与肿瘤淋巴结转移(P值分别为0.003和<0.001)及高TNM分期(P值分别为0.003和0.013)呈显著正相关。结论 Skp2和ABCG2蛋白在鼻咽癌组织中表达上调,并与肿瘤恶性临床病理特征有关。Skp2和ABCG2蛋白可能成为鼻咽癌早期诊断的重要标志物及生物靶向治疗的有效靶点之一。
