Hexokinase Ⅱ has been demonstrated to play the role of a key enzyme member in the glycolysis reaction. It catalyzes the conversion of glucose to glucose-6-phosphate, thus committing glucose to the glycolytic pathway....Hexokinase Ⅱ has been demonstrated to play the role of a key enzyme member in the glycolysis reaction. It catalyzes the conversion of glucose to glucose-6-phosphate, thus committing glucose to the glycolytic pathway. In this paper, the partial exons and introns 10, 11, 13 and 14 of the porcine HK2 gene were cloned and sequenced by comparative genomics. Comparative sequencing of three pig breeds revealed ten putative single-nucleotide polymorphisms （SNPs）, one of which in intron 10 with differing bases （G981A） is within the restriction site for enzyme Msp I. Distribution of Msp I -RFLP genotype and allele frequencies among different pig breeds were studied. By association analysis between Msp I PCR- RFLP polymorphism （AA, AB, BB genotypes of HK2 gene intron 10） and some meat quality and carcass traits in F2 group, which was constructed by our laboratory, a significant difference of pig average backfat at rump was found between AB and BB genotypes （P〈0 05） in F2 group. In addition, the pattern of expression of ilK2 in a variety of tissues in pig was also determined using semi-quantitative RT-PCR. The expression of HK2 mRNA was detected only in pig skeletal muscle.展开更多
目的探究三萜皂苷黄芪皂苷Ⅱ(AstragalosideⅡ,ASⅡ)对三阴乳腺癌(Triple nega⁃tive breast cancer,TNBC)细胞增殖及侵袭转移的作用及可能的机制。方法采用MTT法检测黄芪皂苷Ⅱ(0.78μM、1.56μM、3.12μM、6.25μM、12.5μM、25μM、50...目的探究三萜皂苷黄芪皂苷Ⅱ(AstragalosideⅡ,ASⅡ)对三阴乳腺癌(Triple nega⁃tive breast cancer,TNBC)细胞增殖及侵袭转移的作用及可能的机制。方法采用MTT法检测黄芪皂苷Ⅱ(0.78μM、1.56μM、3.12μM、6.25μM、12.5μM、25μM、50μM、100μM)对MDA-MB-231细胞增殖的影响;采用Transwell法及划痕实验分别检测黄芪皂苷Ⅱ对MDA-MB-231细胞转移及侵袭的影响;采用Western blot检测PI3K/AKT/mTOR通路蛋白的表达水平,从而阐明其调控PI3K/AKT/mTOR通路的分子机制。结果25μM黄芪皂苷Ⅱ显著抑制MDA-MB-231细胞增殖,选用未影响细胞增殖的药物浓度3.12μM、6.25μM、12.5μM作为低、中、高药物浓度;与对照组相比,黄芪皂苷Ⅱ呈浓度依赖性地抑制MDA-MB-231细胞的侵袭与转移;同时,该成分抑制MDA-MB-231细胞PI3K、AKT、mTOR蛋白磷酸化水平。结论黄芪皂苷Ⅱ可能通过调控PI3K/AKT/mTOR通路抑制MDA-MB-231细胞增殖、侵袭及转移。展开更多
基金the National 973 Project, China （G2000016105） National High Technology Development Project （2003AA243030） and the Natural Science Foundation of Hubei Province, China （2005ABA142）.
文摘Hexokinase Ⅱ has been demonstrated to play the role of a key enzyme member in the glycolysis reaction. It catalyzes the conversion of glucose to glucose-6-phosphate, thus committing glucose to the glycolytic pathway. In this paper, the partial exons and introns 10, 11, 13 and 14 of the porcine HK2 gene were cloned and sequenced by comparative genomics. Comparative sequencing of three pig breeds revealed ten putative single-nucleotide polymorphisms （SNPs）, one of which in intron 10 with differing bases （G981A） is within the restriction site for enzyme Msp I. Distribution of Msp I -RFLP genotype and allele frequencies among different pig breeds were studied. By association analysis between Msp I PCR- RFLP polymorphism （AA, AB, BB genotypes of HK2 gene intron 10） and some meat quality and carcass traits in F2 group, which was constructed by our laboratory, a significant difference of pig average backfat at rump was found between AB and BB genotypes （P〈0 05） in F2 group. In addition, the pattern of expression of ilK2 in a variety of tissues in pig was also determined using semi-quantitative RT-PCR. The expression of HK2 mRNA was detected only in pig skeletal muscle.
文摘目的探究三萜皂苷黄芪皂苷Ⅱ(AstragalosideⅡ,ASⅡ)对三阴乳腺癌(Triple nega⁃tive breast cancer,TNBC)细胞增殖及侵袭转移的作用及可能的机制。方法采用MTT法检测黄芪皂苷Ⅱ(0.78μM、1.56μM、3.12μM、6.25μM、12.5μM、25μM、50μM、100μM)对MDA-MB-231细胞增殖的影响;采用Transwell法及划痕实验分别检测黄芪皂苷Ⅱ对MDA-MB-231细胞转移及侵袭的影响;采用Western blot检测PI3K/AKT/mTOR通路蛋白的表达水平,从而阐明其调控PI3K/AKT/mTOR通路的分子机制。结果25μM黄芪皂苷Ⅱ显著抑制MDA-MB-231细胞增殖,选用未影响细胞增殖的药物浓度3.12μM、6.25μM、12.5μM作为低、中、高药物浓度;与对照组相比,黄芪皂苷Ⅱ呈浓度依赖性地抑制MDA-MB-231细胞的侵袭与转移;同时,该成分抑制MDA-MB-231细胞PI3K、AKT、mTOR蛋白磷酸化水平。结论黄芪皂苷Ⅱ可能通过调控PI3K/AKT/mTOR通路抑制MDA-MB-231细胞增殖、侵袭及转移。
