Objective: Tumor cells rely heavily on glycolysis regardless of oxygen tension, a phenomenon called the Warburg effect. Hexokinase II(HKII) catalyzes the first irreversible step of glycolysis and is often overexpresse...Objective: Tumor cells rely heavily on glycolysis regardless of oxygen tension, a phenomenon called the Warburg effect. Hexokinase II(HKII) catalyzes the first irreversible step of glycolysis and is often overexpressed in tumor cells. Mitochondrial HKII couples glycolysis and oxidative phosphorylation while maintaining mitochondrial membrane integrity. In this study, we investigated the role of HKII in promoting the Warburg effect in cancer cells.Methods: HKII-mediated phosphorylation of the alpha subunit of pyruvate dehydrogenase(PDHA1) was tested in HEK293 T cells and clear cell renal cell carcinoma(cc RCC) specimens using gene knockdown, western blotting,immunohistochemistry, and immunofluorescence.Results: It was determined that HKII could not only transform glucose into glucose-6-phosphate, but also transfer the phosphate group of ATP onto PDHA1. In addition, it was found that HKII increased the phosphorylation of Ser293 on PDHA1, decreasing pyruvate dehydrogenase(PDH) complex activity and thus rerouting the metabolic pathway and promoting the Warburg effect. The overexpression of HKII correlated with the phosphorylation of PDHA1 and disease progression in cc RCC.Conclusions: The data presented here suggest that HKII is an important biomarker in the evaluation and treatment of cancer.展开更多
A hexokinase gene named MdHXK1(MDP0000309677) was cloned from ‘Gala' apple(Malus × domestica Borkh.). Sequence analysis showed that the MdHXK1 gene was 1 497 bp long and encoded 499 amino acids. The predicte...A hexokinase gene named MdHXK1(MDP0000309677) was cloned from ‘Gala' apple(Malus × domestica Borkh.). Sequence analysis showed that the MdHXK1 gene was 1 497 bp long and encoded 499 amino acids. The predicted molecular mass of this protein was 54.05 kD, and the pI was 5.76. A phylogenetic tree indicated apple MdHXK1 exhibited the highest sequence similarity to Pyrus bretschneideri PbHXK1. Analysis of the functional domain showed that the MdHXK1 protein included two conserved kinase domains. The prediction of subcellular localization suggested that the MdHXK1 protein was mainly localized in the cytoplasm. There was an indication that MdHXK1 existed as one copy in the apple genome by Southern blotting. Silico analysis suggested that the promoter sequence contained several typical cis-acting elements, including defense, sugar signaling and phytohormone responsive elements. Quantitative real-time PCR analysis demonstrated that the MdHXK1 gene was mainly expressed in stem and flower tissues. During the development of apple fruits, the expression of the MdHXK1 gene initially increased and then decreased. The changes on Glc phosphorylation relative activity and glucose concentration showed the same trend. In addition, the expression of this gene was induced by salt stress, low temperature, and abscisic acid(ABA). Finally, we obtained and purified the fused MdHXK1 protein by recombinant prokaryotic expression. Studies have demonstrated that MdHXK1 may participate in sugar metabolism in apple fruits. Enzyme encoded by MdHXK1 is a key factor in the mediation of sugar accumulation. Recently,researchers on hexokinase at home and abroad mainly focused on model plants, such as Arabidopsis, tobacco and rice, but orchard fruit like apple were underresearched. Our research established the foundation for the further study of the functions of MdHXK1.展开更多
Glucose plays a vital part in milk protein synthesis through the mTOR signaling pathway in bovine mammary epithelial cells(BMEC).The objectives of this study were to determine how glucose affects hexokinase(HK)activit...Glucose plays a vital part in milk protein synthesis through the mTOR signaling pathway in bovine mammary epithelial cells(BMEC).The objectives of this study were to determine how glucose affects hexokinase(HK)activity in BMEC and investigate the regulatory effect of HK in kappa casein(CSN3)synthesis via the mechanistic target of rapamycin complex 1(mTORC1)signaling pathway in BMEC.For this,HK1 and HK2 were knocked out in BMEC using the CRISPR/Cas9 system.The gene and protein expression,glucose uptake,and cell proliferation were measured.We found that glucose uptake,cell proliferation,CSN3 gene expression levels,and expression of HK1 and HK2 increased with increasing glucose concentrations.Notably,glucose uptake was significantly reduced in HK2 knockout(HK2KO)BMEC treated with 17.5 mM glucose.Moreover,under the same glucose treatment conditions,the proliferative ability and abundance of CSN3 were significantly diminished in both HK1 knockout(HK1KO)and HK2KO BMEC compared with that in wild-type BEMC.We further observed that the phosphorylation levels of ribosome protein subunit 6 kinase 1(S6K1)were reduced in HK1KO and HK2KO BMEC following treatment with 17.5 mM glucose.As expected,the levels of glucose-6-phosphate and the m RNA expression levels of glycolysis-related genes were decreased in both HK1KO and HK2KO BMEC following glucose treatment.These results indicated that the knockout of HK1 and HK2 inhibited cell proliferation and CSN3 expression in BMEC under glucose treatment,which may be associated with the inactivation of the S6K1 and inhibition of glycolysis.展开更多
Objective To investigate the regulatory relationship of Protein Phosphatase 2 Regulatory Subunit B"Alpha(PPP2R3A)and hexokinase 1(HK1)in glycolysis of hepatocellular carcinoma(HCC).Methods In HepG2 and Huh7 cells...Objective To investigate the regulatory relationship of Protein Phosphatase 2 Regulatory Subunit B"Alpha(PPP2R3A)and hexokinase 1(HK1)in glycolysis of hepatocellular carcinoma(HCC).Methods In HepG2 and Huh7 cells,PPP2R3A expression was silenced by small interfering RNA(siRNA)and overexpression by plasmid transfection.The PPP2R3A-related genes were searched by RNA sequencing.Glycolysis levels were measured by glucose uptake and lactate production.QRT-PCR,ELISA,western blot and immunofluorescence assay were performed to detect the changes of PPP2R3A and HK1.Cell proliferation,migration and invasion assay were used to study the roles of HK1 regulation by PPP2R3A.Results RNA sequencing data revealed that PPP2R3A siRNA significantly downregulated the expression of HK1.PPP2R3A gene overexpression promotes,while gene silencing suppresses,the level of HK1 and glycolysis in HCC cells.In HCC tissue samples,PPP2R3A and HK1 were colocalized in the cytoplasm,and their expression showed a positive correlation.HK1 inhibition abrogated the promotion of glycolysis,proliferation,migration and invasion by PPP2R3A overexpression in liver cancer cells.Conclusion Our findings showed the correlation of PPP2R3A and HK1 in the glycolysis of HCC,which reveals a new mechanism for the oncogenic roles of PPP2R3A in cancer.展开更多
OBJECTIVE:To investigate the effect of Ruyanneixiao cream(RYNX) on the expression of hypoxia inducible factor-1α(HIF-1α), hexokinase 2(HK2),phosphofructokinase(PFK), and pyruvate kinase M2(PKM2) mRNA and protein in ...OBJECTIVE:To investigate the effect of Ruyanneixiao cream(RYNX) on the expression of hypoxia inducible factor-1α(HIF-1α), hexokinase 2(HK2),phosphofructokinase(PFK), and pyruvate kinase M2(PKM2) mRNA and protein in MCF-10 AT cells and in an animal model of precancerous mammary lesions.METHODS:Following treatment of MCF-10 AT cells with RYNX, tamoxifen(TAM) and YC-1 for 48 h,HIF-1α, HK2, PFK, PKM2 mRNA and protein expression was analyzed.Fifty female SD rats were randomly divided into control, model, TAM, and highand low-dose RYNX groups, with 10 rats in each group.A precancerous mammary lesion model was established for all groups except the control group.High-and low-dose RYNX cream containing TAM was coated on the breasts of animals in the corresponding groups.The rat mammary tissue was removed in the 10 th week and HIF-1α, HK2, PFK,PKM2 mRNA and protein was analyzed.RESULTS:In vitro analyses demonstrated that, compared with the matrix group, HIF-1α, HK2, PFK,PKM2 mRNA and protein expression was significantly decreased in the RYNX group(P < 0.05).Compared with the YC-1 + RYNX group, HK2, PFK,and PKM2 protein expression was significantly reduced in the RYNX group.HIF-1α, HK2, PFK, and PKM2 protein expression was increased significantly in the model group(P < 0.05) compared with the control group, while HIF-1α, HK2, PFK, and PKM2 mRNA and protein expression was significantly decreased in both the high-and low-dose RYNX groups(P < 0.05), with the effect being greater in the high-dose group.CONCLUSION:RYNX can block precancerous breast lesions by decreasing the expression of HK2,PFK, and PKM2 mRNA and protein via inhibition of HIF-1α mRNA and protein overexpression in a dose-dependent manner.展开更多
目的分析缺血性视网膜静脉阻塞继发黄斑水肿(RVO-ME)患者基线血清己糖激酶1抗体滴度与抗血管内皮生长因子(VEGF)治疗后视力改善的相关性。方法招募2017年6月至2020年2月在首都医科大学宣武医院确诊为缺血性RVO-ME并接受初始抗VEGF治疗...目的分析缺血性视网膜静脉阻塞继发黄斑水肿(RVO-ME)患者基线血清己糖激酶1抗体滴度与抗血管内皮生长因子(VEGF)治疗后视力改善的相关性。方法招募2017年6月至2020年2月在首都医科大学宣武医院确诊为缺血性RVO-ME并接受初始抗VEGF治疗的53例患者,其中缺血性视网膜中央静脉阻塞(CRVO)23例(CRVO组),缺血性视网膜分支静脉阻塞(BRVO)30例(BRVO组)。另选取该院同期30例行超声乳化的白内障患者作为对照组。研究对象行基线血清己糖激酶1抗体滴度检测、眼科常规检查和光学相干断层成像(OCT)检查。所有RVO-ME患者按照“3+按需治疗方案(pro re nata,PRN)”向玻璃体内注射抗VEGF药物治疗。随访12个月,采用多元线性回归分析缺血性RVO-ME患者抗VEGF治疗后视力改善的影响因素。结果CRVO组基线logMAR BCVA高于对照组和BRVO组,CRVO组和BRVO组基线CRT、基线血清己糖激酶1抗体滴度高于对照组,且CRVO组基线CRT、基线血清己糖激酶1抗体滴度高于BRVO组,差异有统计学意义(P<0.05)。RVO-ME患者基线血清己糖激酶1抗体滴度与随访6个月(r=0.377,P=0.005)、9个月(r=0.362,P=0.008)和12个月(r=0.465,P<0.001)时BCVA改善呈正相关,与随访12个月时中断EZ横向长度减少值(r=0.401,P=0.001)呈正相关。多元线性回归分析结果显示,基线logMAR BCVA、基线血清己糖激酶1抗体滴度是缺血性RVO-ME患者抗VEGF治疗随访12个月时BCVA改善的影响因素(P<0.05)。结论己糖激酶1抗体作为一种新的血清生物标志物,与缺血性RVO-ME患者抗VEGF治疗后的视力改善相关。展开更多
目的 探讨己糖激酶结构域成分1(HKDC1)对人结直肠癌细胞增殖、迁移、侵袭能力的影响。方法 HT-29、SW480细胞进行瞬时转染,设置HKDC1-si RNA组、阴性对照组及空白组。同时收集2020年3月至2022年5月河北北方学院附属第一医院接受手术的5...目的 探讨己糖激酶结构域成分1(HKDC1)对人结直肠癌细胞增殖、迁移、侵袭能力的影响。方法 HT-29、SW480细胞进行瞬时转染,设置HKDC1-si RNA组、阴性对照组及空白组。同时收集2020年3月至2022年5月河北北方学院附属第一医院接受手术的50例患者的结直肠癌组织和癌旁正常组织。RT-q PCR实验检测HKDC1 m RNA表达;Western blot实验检测HKDC1蛋白表达;CCK-8实验、划痕实验和Transwell实验检测细胞增殖、迁移和侵袭能力。结果 HT-29细胞HKDC1 m RNA的表达量高于SW480细胞(P<0.05),故选取HT-29细胞株用于后续实验。结直肠癌组织中HKDC1 m RNA、光密度值高于癌旁正常组织(P<0.05)。空白组与阴性对照组HKDC1 m RNA、蛋白表达水平比较,差异无统计学意义(P>0.05);HKDC1-si RNA组HKDC1 m RNA、蛋白表达水平低于阴性对照组(P<0.05)。空白组与阴性对照组光密度值比较,差异无统计学意义(P>0.05);HKDC1-si RNA组光密度值低于阴性对照组(P<0.05)。空白组与阴性对照组细胞迁移率比较,差异无统计学意义(P>0.05);HKDC1-si RNA组细胞迁移率低于阴性对照组(P<0.05)。空白组与阴性对照组细胞迁移数目比较,差异无统计学意义(P>0.05);HKDC1-si RNA组细胞迁移数目低于阴性对照组(P<0.05)。结论 HKDC1在结直肠癌组织及细胞中高表达,可促进结直肠癌细胞的增殖、迁移及侵袭能力,有望成为结直肠癌基因治疗的新靶点。展开更多
目的:探讨香加皮提取物杠柳苷(Periplocin from cortex periplocae,CPP)对人类急性淋巴细胞白血病(ALL)细胞株Jurkat的抑制作用及作用机制。方法:CASY快速细胞计数法检测CPP对人Jurkat的敏感性。将AnnexinV/PI双染试剂盒染色结合流式细...目的:探讨香加皮提取物杠柳苷(Periplocin from cortex periplocae,CPP)对人类急性淋巴细胞白血病(ALL)细胞株Jurkat的抑制作用及作用机制。方法:CASY快速细胞计数法检测CPP对人Jurkat的敏感性。将AnnexinV/PI双染试剂盒染色结合流式细胞术检测CPP作用不同时间,诱导Jurkat凋亡坏死细胞比例。采用CPP作用Jurkat 12h,实时定量PCR检测CPP对葡萄糖转运蛋白(Glut)、人己糖激酶-II(HK-II)及缺氧诱发因子(HIF-1α)基因拷贝数的影响。结果:CPP对Jurkat有显著的细胞毒作用,其半数抑制浓度为(48.33±6.24)nM。CPP能诱导Jurkat凋亡,凋亡率随着作用时间的延长及剂量的增加而明显增高,呈时间、剂量依赖性。CPP作用于Jurkat细胞12h后,CPP不能抑制HIF-1αmRNA的表达,而能显著抑制糖酵解相关的Glut、HK-II基因的表达。结论:CPP能有效抑制ALL细胞株Jurkat的生长,其机制与诱发凋亡、抑制糖酵解密切相关。展开更多
文摘Objective: Tumor cells rely heavily on glycolysis regardless of oxygen tension, a phenomenon called the Warburg effect. Hexokinase II(HKII) catalyzes the first irreversible step of glycolysis and is often overexpressed in tumor cells. Mitochondrial HKII couples glycolysis and oxidative phosphorylation while maintaining mitochondrial membrane integrity. In this study, we investigated the role of HKII in promoting the Warburg effect in cancer cells.Methods: HKII-mediated phosphorylation of the alpha subunit of pyruvate dehydrogenase(PDHA1) was tested in HEK293 T cells and clear cell renal cell carcinoma(cc RCC) specimens using gene knockdown, western blotting,immunohistochemistry, and immunofluorescence.Results: It was determined that HKII could not only transform glucose into glucose-6-phosphate, but also transfer the phosphate group of ATP onto PDHA1. In addition, it was found that HKII increased the phosphorylation of Ser293 on PDHA1, decreasing pyruvate dehydrogenase(PDH) complex activity and thus rerouting the metabolic pathway and promoting the Warburg effect. The overexpression of HKII correlated with the phosphorylation of PDHA1 and disease progression in cc RCC.Conclusions: The data presented here suggest that HKII is an important biomarker in the evaluation and treatment of cancer.
基金supported by the National Science Foundation for Distinguished Young Scholars of China(Grant No.31325024)Innovation Team Project of Shandong Modern Agricultural Industry Technology System(SDAIT-03-022-03)
文摘A hexokinase gene named MdHXK1(MDP0000309677) was cloned from ‘Gala' apple(Malus × domestica Borkh.). Sequence analysis showed that the MdHXK1 gene was 1 497 bp long and encoded 499 amino acids. The predicted molecular mass of this protein was 54.05 kD, and the pI was 5.76. A phylogenetic tree indicated apple MdHXK1 exhibited the highest sequence similarity to Pyrus bretschneideri PbHXK1. Analysis of the functional domain showed that the MdHXK1 protein included two conserved kinase domains. The prediction of subcellular localization suggested that the MdHXK1 protein was mainly localized in the cytoplasm. There was an indication that MdHXK1 existed as one copy in the apple genome by Southern blotting. Silico analysis suggested that the promoter sequence contained several typical cis-acting elements, including defense, sugar signaling and phytohormone responsive elements. Quantitative real-time PCR analysis demonstrated that the MdHXK1 gene was mainly expressed in stem and flower tissues. During the development of apple fruits, the expression of the MdHXK1 gene initially increased and then decreased. The changes on Glc phosphorylation relative activity and glucose concentration showed the same trend. In addition, the expression of this gene was induced by salt stress, low temperature, and abscisic acid(ABA). Finally, we obtained and purified the fused MdHXK1 protein by recombinant prokaryotic expression. Studies have demonstrated that MdHXK1 may participate in sugar metabolism in apple fruits. Enzyme encoded by MdHXK1 is a key factor in the mediation of sugar accumulation. Recently,researchers on hexokinase at home and abroad mainly focused on model plants, such as Arabidopsis, tobacco and rice, but orchard fruit like apple were underresearched. Our research established the foundation for the further study of the functions of MdHXK1.
基金supported by the Development Project of China(2017YFD0502104-3)the China Agriculture Research System(CARS-36)the National Natural Science Foundation of China(No.31972589)
文摘Glucose plays a vital part in milk protein synthesis through the mTOR signaling pathway in bovine mammary epithelial cells(BMEC).The objectives of this study were to determine how glucose affects hexokinase(HK)activity in BMEC and investigate the regulatory effect of HK in kappa casein(CSN3)synthesis via the mechanistic target of rapamycin complex 1(mTORC1)signaling pathway in BMEC.For this,HK1 and HK2 were knocked out in BMEC using the CRISPR/Cas9 system.The gene and protein expression,glucose uptake,and cell proliferation were measured.We found that glucose uptake,cell proliferation,CSN3 gene expression levels,and expression of HK1 and HK2 increased with increasing glucose concentrations.Notably,glucose uptake was significantly reduced in HK2 knockout(HK2KO)BMEC treated with 17.5 mM glucose.Moreover,under the same glucose treatment conditions,the proliferative ability and abundance of CSN3 were significantly diminished in both HK1 knockout(HK1KO)and HK2KO BMEC compared with that in wild-type BEMC.We further observed that the phosphorylation levels of ribosome protein subunit 6 kinase 1(S6K1)were reduced in HK1KO and HK2KO BMEC following treatment with 17.5 mM glucose.As expected,the levels of glucose-6-phosphate and the m RNA expression levels of glycolysis-related genes were decreased in both HK1KO and HK2KO BMEC following glucose treatment.These results indicated that the knockout of HK1 and HK2 inhibited cell proliferation and CSN3 expression in BMEC under glucose treatment,which may be associated with the inactivation of the S6K1 and inhibition of glycolysis.
基金supported by National Natural Science Foundation of China [81372595]
文摘Objective To investigate the regulatory relationship of Protein Phosphatase 2 Regulatory Subunit B"Alpha(PPP2R3A)and hexokinase 1(HK1)in glycolysis of hepatocellular carcinoma(HCC).Methods In HepG2 and Huh7 cells,PPP2R3A expression was silenced by small interfering RNA(siRNA)and overexpression by plasmid transfection.The PPP2R3A-related genes were searched by RNA sequencing.Glycolysis levels were measured by glucose uptake and lactate production.QRT-PCR,ELISA,western blot and immunofluorescence assay were performed to detect the changes of PPP2R3A and HK1.Cell proliferation,migration and invasion assay were used to study the roles of HK1 regulation by PPP2R3A.Results RNA sequencing data revealed that PPP2R3A siRNA significantly downregulated the expression of HK1.PPP2R3A gene overexpression promotes,while gene silencing suppresses,the level of HK1 and glycolysis in HCC cells.In HCC tissue samples,PPP2R3A and HK1 were colocalized in the cytoplasm,and their expression showed a positive correlation.HK1 inhibition abrogated the promotion of glycolysis,proliferation,migration and invasion by PPP2R3A overexpression in liver cancer cells.Conclusion Our findings showed the correlation of PPP2R3A and HK1 in the glycolysis of HCC,which reveals a new mechanism for the oncogenic roles of PPP2R3A in cancer.
基金Supported by National Natural Science Foundation Project of China(No.81673979,81473688,81173265)Traditional Chinese Medicine Administration Project of Guangdong Province,China(No.20141070)+3 种基金the Science and Technology Program of Guangdong,China(No.2014A020212672,2014A020210015)the Natural Science Foundation of Guangdong,China(No.2016A030313114,2015A030313333)Scientific Research Cultivation and Innovation of Jinan University Special Fund for Basic Scientific Research of Central University(No.21615464,21615412)the New Century Talent Support Program by the Ministry of Education(No.NCET-13-0827)
文摘OBJECTIVE:To investigate the effect of Ruyanneixiao cream(RYNX) on the expression of hypoxia inducible factor-1α(HIF-1α), hexokinase 2(HK2),phosphofructokinase(PFK), and pyruvate kinase M2(PKM2) mRNA and protein in MCF-10 AT cells and in an animal model of precancerous mammary lesions.METHODS:Following treatment of MCF-10 AT cells with RYNX, tamoxifen(TAM) and YC-1 for 48 h,HIF-1α, HK2, PFK, PKM2 mRNA and protein expression was analyzed.Fifty female SD rats were randomly divided into control, model, TAM, and highand low-dose RYNX groups, with 10 rats in each group.A precancerous mammary lesion model was established for all groups except the control group.High-and low-dose RYNX cream containing TAM was coated on the breasts of animals in the corresponding groups.The rat mammary tissue was removed in the 10 th week and HIF-1α, HK2, PFK,PKM2 mRNA and protein was analyzed.RESULTS:In vitro analyses demonstrated that, compared with the matrix group, HIF-1α, HK2, PFK,PKM2 mRNA and protein expression was significantly decreased in the RYNX group(P < 0.05).Compared with the YC-1 + RYNX group, HK2, PFK,and PKM2 protein expression was significantly reduced in the RYNX group.HIF-1α, HK2, PFK, and PKM2 protein expression was increased significantly in the model group(P < 0.05) compared with the control group, while HIF-1α, HK2, PFK, and PKM2 mRNA and protein expression was significantly decreased in both the high-and low-dose RYNX groups(P < 0.05), with the effect being greater in the high-dose group.CONCLUSION:RYNX can block precancerous breast lesions by decreasing the expression of HK2,PFK, and PKM2 mRNA and protein via inhibition of HIF-1α mRNA and protein overexpression in a dose-dependent manner.
文摘目的分析缺血性视网膜静脉阻塞继发黄斑水肿(RVO-ME)患者基线血清己糖激酶1抗体滴度与抗血管内皮生长因子(VEGF)治疗后视力改善的相关性。方法招募2017年6月至2020年2月在首都医科大学宣武医院确诊为缺血性RVO-ME并接受初始抗VEGF治疗的53例患者,其中缺血性视网膜中央静脉阻塞(CRVO)23例(CRVO组),缺血性视网膜分支静脉阻塞(BRVO)30例(BRVO组)。另选取该院同期30例行超声乳化的白内障患者作为对照组。研究对象行基线血清己糖激酶1抗体滴度检测、眼科常规检查和光学相干断层成像(OCT)检查。所有RVO-ME患者按照“3+按需治疗方案(pro re nata,PRN)”向玻璃体内注射抗VEGF药物治疗。随访12个月,采用多元线性回归分析缺血性RVO-ME患者抗VEGF治疗后视力改善的影响因素。结果CRVO组基线logMAR BCVA高于对照组和BRVO组,CRVO组和BRVO组基线CRT、基线血清己糖激酶1抗体滴度高于对照组,且CRVO组基线CRT、基线血清己糖激酶1抗体滴度高于BRVO组,差异有统计学意义(P<0.05)。RVO-ME患者基线血清己糖激酶1抗体滴度与随访6个月(r=0.377,P=0.005)、9个月(r=0.362,P=0.008)和12个月(r=0.465,P<0.001)时BCVA改善呈正相关,与随访12个月时中断EZ横向长度减少值(r=0.401,P=0.001)呈正相关。多元线性回归分析结果显示,基线logMAR BCVA、基线血清己糖激酶1抗体滴度是缺血性RVO-ME患者抗VEGF治疗随访12个月时BCVA改善的影响因素(P<0.05)。结论己糖激酶1抗体作为一种新的血清生物标志物,与缺血性RVO-ME患者抗VEGF治疗后的视力改善相关。
文摘目的 探讨己糖激酶结构域成分1(HKDC1)对人结直肠癌细胞增殖、迁移、侵袭能力的影响。方法 HT-29、SW480细胞进行瞬时转染,设置HKDC1-si RNA组、阴性对照组及空白组。同时收集2020年3月至2022年5月河北北方学院附属第一医院接受手术的50例患者的结直肠癌组织和癌旁正常组织。RT-q PCR实验检测HKDC1 m RNA表达;Western blot实验检测HKDC1蛋白表达;CCK-8实验、划痕实验和Transwell实验检测细胞增殖、迁移和侵袭能力。结果 HT-29细胞HKDC1 m RNA的表达量高于SW480细胞(P<0.05),故选取HT-29细胞株用于后续实验。结直肠癌组织中HKDC1 m RNA、光密度值高于癌旁正常组织(P<0.05)。空白组与阴性对照组HKDC1 m RNA、蛋白表达水平比较,差异无统计学意义(P>0.05);HKDC1-si RNA组HKDC1 m RNA、蛋白表达水平低于阴性对照组(P<0.05)。空白组与阴性对照组光密度值比较,差异无统计学意义(P>0.05);HKDC1-si RNA组光密度值低于阴性对照组(P<0.05)。空白组与阴性对照组细胞迁移率比较,差异无统计学意义(P>0.05);HKDC1-si RNA组细胞迁移率低于阴性对照组(P<0.05)。空白组与阴性对照组细胞迁移数目比较,差异无统计学意义(P>0.05);HKDC1-si RNA组细胞迁移数目低于阴性对照组(P<0.05)。结论 HKDC1在结直肠癌组织及细胞中高表达,可促进结直肠癌细胞的增殖、迁移及侵袭能力,有望成为结直肠癌基因治疗的新靶点。
文摘目的:探讨香加皮提取物杠柳苷(Periplocin from cortex periplocae,CPP)对人类急性淋巴细胞白血病(ALL)细胞株Jurkat的抑制作用及作用机制。方法:CASY快速细胞计数法检测CPP对人Jurkat的敏感性。将AnnexinV/PI双染试剂盒染色结合流式细胞术检测CPP作用不同时间,诱导Jurkat凋亡坏死细胞比例。采用CPP作用Jurkat 12h,实时定量PCR检测CPP对葡萄糖转运蛋白(Glut)、人己糖激酶-II(HK-II)及缺氧诱发因子(HIF-1α)基因拷贝数的影响。结果:CPP对Jurkat有显著的细胞毒作用,其半数抑制浓度为(48.33±6.24)nM。CPP能诱导Jurkat凋亡,凋亡率随着作用时间的延长及剂量的增加而明显增高,呈时间、剂量依赖性。CPP作用于Jurkat细胞12h后,CPP不能抑制HIF-1αmRNA的表达,而能显著抑制糖酵解相关的Glut、HK-II基因的表达。结论:CPP能有效抑制ALL细胞株Jurkat的生长,其机制与诱发凋亡、抑制糖酵解密切相关。
