目的:探究磷酸甘油酸变位酶1(phosphoglycerate mutase 1,PGAM1)变构抑制剂能否提高结肠癌细胞对奥沙利铂(oxaliplatin,OXA)的敏感性。方法:癌症药物敏感性基因组学(genomics of drug sensitivity in cancer,GDSC)和癌细胞系百科全书数...目的:探究磷酸甘油酸变位酶1(phosphoglycerate mutase 1,PGAM1)变构抑制剂能否提高结肠癌细胞对奥沙利铂(oxaliplatin,OXA)的敏感性。方法:癌症药物敏感性基因组学(genomics of drug sensitivity in cancer,GDSC)和癌细胞系百科全书数据库(cancer cell line encyclopedia,CCLE)分析四种共识分子亚型(consensus molecular subgroups,CMS)结肠癌细胞对OXA敏感性和PGAM1的表达水平;采用CCK8法和Annexin V/PI双染法,探究PGAM1变构抑制剂HKB99对人源结肠癌细胞增殖和凋亡的作用;采用CCK8法分析HKB99联合OXA对结肠癌增殖的作用,并进行协同系数分析。结果:首先,GDSC和CCLE数据库分析结果显示,相对于CMS2和CMS3亚型,CMS1和CMS4亚型结肠癌细胞对OXA敏感的细胞比例降低,PGAM1表达升高,具有显著性差异(P<0.05)。进一步研究表明,HKB99显著抑制不同CMS类型的人源结肠癌SW480、SCUN2B和DLD-1细胞的增殖,其IC50均为1μM左右;Annexin V/PI双染法结果显示,与对照组相比,HKB99处理组DLD-1细胞的凋亡水平显著上升(P<0.01)。最后,CCK8法检测显示,与OXA单加药组相比,1.5μM的HKB99显著提高结肠癌DLD-1细胞对OXA的敏感性(IC_(50):3.26 vs0.37μM);协同系数分析结果显示,OXA在浓度1.56μM和25μM之间与HKB99的协同系数小于1。结论:本研究发现相对于CMS2和CMS3,CMS1和CMS4结肠癌细胞对OXA敏感性较低且PGAM1表达水平升高,PGAM1变构抑制剂HKB99增强结肠癌细胞对OXA的敏感性,提示HKB99有望提高结肠癌OXA化疗的敏感性,为患者个体化化疗提供新的治疗策略。展开更多
目的:探讨磷酸甘油酸变位酶1(PGAM1)在乳腺癌及癌旁正常组织中的表达及意义。方法:应用real time PCR,免疫组化法分别检测32例乳腺癌及癌旁正常组织中PGAM1在mRNA及蛋白水平的表达。结果:在收集的32例乳腺癌中PGAM1的表达明显高于癌旁...目的:探讨磷酸甘油酸变位酶1(PGAM1)在乳腺癌及癌旁正常组织中的表达及意义。方法:应用real time PCR,免疫组化法分别检测32例乳腺癌及癌旁正常组织中PGAM1在mRNA及蛋白水平的表达。结果:在收集的32例乳腺癌中PGAM1的表达明显高于癌旁正常组织(P<0.05)。结论:PGAM1在乳腺癌组织中有高表达,为乳腺癌的诊断及治疗提供了新的思路。展开更多
To provide the useful information for the choice of molecular marker used in marker-assisted selection of drought tolerance, it is necessary to find out more candidate genes and fulfill the information gaps in gene ex...To provide the useful information for the choice of molecular marker used in marker-assisted selection of drought tolerance, it is necessary to find out more candidate genes and fulfill the information gaps in gene expression regulation under drought stress. In this study, we isolated four differentially expressed cDNA fragments from leaves of a droughttolerant inbred line by suppression subtractive hybridization and reverse Northern hybridization, and validated their differential expression patterns among six inbred lines with different drought tolerance in response to drought stress by quantitative real-time PCR. Sequence similarity analysis indicated that two of four differentially expressed cDNA showed homology to gene DegP encoding trypsin-like serine protease, and gene PGAM-i encoding cofactor-independent phosphoglyceromutase, respectively. Expressions of the genes corresponding to four cDNA fragments was decreased at 6 h after drought stress treatment in most of the six inbred lines, and then returned to the control level with further stress in three of the tolerant inbred lines. The expression of the gene PGAM-i and the genes corresponding to fragments E4 and F4 were increased to a high level in tolerant inbred line 81565. In the two drought-sensitive inbred lines （Dan340 and ES40）, the expression of these genes was still down-regulated. The probable mechanisms of these genes in response to drought stress were discussed. These results indicated that the drought-tolerant inbred lines upregulated the expression of the drought-tolerant candidate genes, in contrast, drought-sensitive inbred lines downregulated the expression of the genes.展开更多
文摘目的:探究磷酸甘油酸变位酶1(phosphoglycerate mutase 1,PGAM1)变构抑制剂能否提高结肠癌细胞对奥沙利铂(oxaliplatin,OXA)的敏感性。方法:癌症药物敏感性基因组学(genomics of drug sensitivity in cancer,GDSC)和癌细胞系百科全书数据库(cancer cell line encyclopedia,CCLE)分析四种共识分子亚型(consensus molecular subgroups,CMS)结肠癌细胞对OXA敏感性和PGAM1的表达水平;采用CCK8法和Annexin V/PI双染法,探究PGAM1变构抑制剂HKB99对人源结肠癌细胞增殖和凋亡的作用;采用CCK8法分析HKB99联合OXA对结肠癌增殖的作用,并进行协同系数分析。结果:首先,GDSC和CCLE数据库分析结果显示,相对于CMS2和CMS3亚型,CMS1和CMS4亚型结肠癌细胞对OXA敏感的细胞比例降低,PGAM1表达升高,具有显著性差异(P<0.05)。进一步研究表明,HKB99显著抑制不同CMS类型的人源结肠癌SW480、SCUN2B和DLD-1细胞的增殖,其IC50均为1μM左右;Annexin V/PI双染法结果显示,与对照组相比,HKB99处理组DLD-1细胞的凋亡水平显著上升(P<0.01)。最后,CCK8法检测显示,与OXA单加药组相比,1.5μM的HKB99显著提高结肠癌DLD-1细胞对OXA的敏感性(IC_(50):3.26 vs0.37μM);协同系数分析结果显示,OXA在浓度1.56μM和25μM之间与HKB99的协同系数小于1。结论:本研究发现相对于CMS2和CMS3,CMS1和CMS4结肠癌细胞对OXA敏感性较低且PGAM1表达水平升高,PGAM1变构抑制剂HKB99增强结肠癌细胞对OXA的敏感性,提示HKB99有望提高结肠癌OXA化疗的敏感性,为患者个体化化疗提供新的治疗策略。
文摘目的:探讨磷酸甘油酸变位酶1(PGAM1)在乳腺癌及癌旁正常组织中的表达及意义。方法:应用real time PCR,免疫组化法分别检测32例乳腺癌及癌旁正常组织中PGAM1在mRNA及蛋白水平的表达。结果:在收集的32例乳腺癌中PGAM1的表达明显高于癌旁正常组织(P<0.05)。结论:PGAM1在乳腺癌组织中有高表达,为乳腺癌的诊断及治疗提供了新的思路。
基金supported by the National Natural Science Foundation of China (30571172, 30671309, and30721140554)Rockefeller Foundation (2004 FS 047)the Program for Changjiang Scholar and Innovative Research Team in Universities, China (PCSIRT,IRT0453)
文摘To provide the useful information for the choice of molecular marker used in marker-assisted selection of drought tolerance, it is necessary to find out more candidate genes and fulfill the information gaps in gene expression regulation under drought stress. In this study, we isolated four differentially expressed cDNA fragments from leaves of a droughttolerant inbred line by suppression subtractive hybridization and reverse Northern hybridization, and validated their differential expression patterns among six inbred lines with different drought tolerance in response to drought stress by quantitative real-time PCR. Sequence similarity analysis indicated that two of four differentially expressed cDNA showed homology to gene DegP encoding trypsin-like serine protease, and gene PGAM-i encoding cofactor-independent phosphoglyceromutase, respectively. Expressions of the genes corresponding to four cDNA fragments was decreased at 6 h after drought stress treatment in most of the six inbred lines, and then returned to the control level with further stress in three of the tolerant inbred lines. The expression of the gene PGAM-i and the genes corresponding to fragments E4 and F4 were increased to a high level in tolerant inbred line 81565. In the two drought-sensitive inbred lines （Dan340 and ES40）, the expression of these genes was still down-regulated. The probable mechanisms of these genes in response to drought stress were discussed. These results indicated that the drought-tolerant inbred lines upregulated the expression of the drought-tolerant candidate genes, in contrast, drought-sensitive inbred lines downregulated the expression of the genes.
