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
Background: More and more chronic kidney disease (CKD) patients are accompanied with hyperuricaemia. As is known, hyperuricaemia is an independent hazard of both cardiovascular diseases (CVD) and chronic kidney diseas...Background: More and more chronic kidney disease (CKD) patients are accompanied with hyperuricaemia. As is known, hyperuricaemia is an independent hazard of both cardiovascular diseases (CVD) and chronic kidney diseases. We aim at identifying Single Nucleotide Polymorphism (SNP) difference of hURAT1 (rs7932775) and ABCG2 (rs3825016) on CKD patient with hyperuricemia and/or gout. Methods: All forty-two CKD patients were divided into two groups: hyperuricemia, and control group. 24 hours urine sample and serum were prepared for testing biochemistry parameters. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method is used to analyze hURAT1 and ABCG2 single nucleotide polymorphisms in different groups. Results: 17 patients have CT SNP of hURAT1 (rs7932775) and 13 patients have CT SNP of ABCG2 (rs3825016) in hyperuricemia group, while only 5 persons and 6 persons have the same mutations in control group respectively. 7 patients have CT SNP of both hURAT1 (rs7932775) and ABCG2 (rs3825016) in hyperuricemia group, while only 2 persons have the same mutations in control group. CT mutation rates of hURAT1 (rs7932775) and ABCG2 (rs3825016) in hyperuricemia group were 60.7% (17/28) and 50% (13/28) respectively, higher than that of control group (35.7% (5/14) and 42.8% (6/14)). What is more, Double SNP mutations in both hURAT1 (rs7932775) and ABCG2 (rs3825016) in hyperuricemia group were 25% (7/28), higher than that of control group (14.2%, 2/14). Conclusion: There are higher mutation rates of CT SNP in hURAT1 (rs7932775) and/or ABCG2 (rs3825016) in hyperuricemia group. We can conclude that hyperuricemia is a high risk factor in progress of CKD, which is necessary to take measures of decreasing serum uric acid to delay CKD progress.展开更多
Objective:To determine the effect of steroidogenic acute regulatory protein(StAR) overexpression on the levels of adenosine triphosphate(ATP)-binding cassette transporter A1(ABCA1) and ATP-binding cassette transporter...Objective:To determine the effect of steroidogenic acute regulatory protein(StAR) overexpression on the levels of adenosine triphosphate(ATP)-binding cassette transporter A1(ABCA1) and ATP-binding cassette transporter G1(ABCG1) in an endothelial cell line(bEnd.3).Methods:The StAR gene was induced in bEnd.3 cells with adenovirus infection.The infection efficiency was detected by fluorescence activated cell sorter(FACS) and fluorescence microscopy.The expressions of StAR gene and protein levels were detected by real-time polymerase chain reaction(PCR) and Western blot.The gene and protein levels of ABCA1 and ABCG1 were detected by real-time PCR and Western blot after StAR overexpression.Results:The result shows that StAR was successfully overexpressed in bEnd.3 cells by adenovirus infection.The mRNA and protein expressions of ABCA1 and ABCG1 were greatly increased by StAR overexpression in bEnd.3 cells.Conclusion:Overexpression of StAR increases ABCA1 and ABCG1 expressions in endothelial cells.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Background: More and more chronic kidney disease (CKD) patients are accompanied with hyperuricaemia. As is known, hyperuricaemia is an independent hazard of both cardiovascular diseases (CVD) and chronic kidney diseases. We aim at identifying Single Nucleotide Polymorphism (SNP) difference of hURAT1 (rs7932775) and ABCG2 (rs3825016) on CKD patient with hyperuricemia and/or gout. Methods: All forty-two CKD patients were divided into two groups: hyperuricemia, and control group. 24 hours urine sample and serum were prepared for testing biochemistry parameters. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method is used to analyze hURAT1 and ABCG2 single nucleotide polymorphisms in different groups. Results: 17 patients have CT SNP of hURAT1 (rs7932775) and 13 patients have CT SNP of ABCG2 (rs3825016) in hyperuricemia group, while only 5 persons and 6 persons have the same mutations in control group respectively. 7 patients have CT SNP of both hURAT1 (rs7932775) and ABCG2 (rs3825016) in hyperuricemia group, while only 2 persons have the same mutations in control group. CT mutation rates of hURAT1 (rs7932775) and ABCG2 (rs3825016) in hyperuricemia group were 60.7% (17/28) and 50% (13/28) respectively, higher than that of control group (35.7% (5/14) and 42.8% (6/14)). What is more, Double SNP mutations in both hURAT1 (rs7932775) and ABCG2 (rs3825016) in hyperuricemia group were 25% (7/28), higher than that of control group (14.2%, 2/14). Conclusion: There are higher mutation rates of CT SNP in hURAT1 (rs7932775) and/or ABCG2 (rs3825016) in hyperuricemia group. We can conclude that hyperuricemia is a high risk factor in progress of CKD, which is necessary to take measures of decreasing serum uric acid to delay CKD progress.
基金Project (Nos 30871021 and 30900716) supported by the National Natural Science Foundation of China
文摘Objective:To determine the effect of steroidogenic acute regulatory protein(StAR) overexpression on the levels of adenosine triphosphate(ATP)-binding cassette transporter A1(ABCA1) and ATP-binding cassette transporter G1(ABCG1) in an endothelial cell line(bEnd.3).Methods:The StAR gene was induced in bEnd.3 cells with adenovirus infection.The infection efficiency was detected by fluorescence activated cell sorter(FACS) and fluorescence microscopy.The expressions of StAR gene and protein levels were detected by real-time polymerase chain reaction(PCR) and Western blot.The gene and protein levels of ABCA1 and ABCG1 were detected by real-time PCR and Western blot after StAR overexpression.Results:The result shows that StAR was successfully overexpressed in bEnd.3 cells by adenovirus infection.The mRNA and protein expressions of ABCA1 and ABCG1 were greatly increased by StAR overexpression in bEnd.3 cells.Conclusion:Overexpression of StAR increases ABCA1 and ABCG1 expressions in endothelial cells.
