Objective Hepatocellular carcinoma(HCC)is the third leading cause of cancer-associated death worldwide.As a first-line drug for advanced HCC treatment,lenvatinib faces a significant hurdle due to the development of bo...Objective Hepatocellular carcinoma(HCC)is the third leading cause of cancer-associated death worldwide.As a first-line drug for advanced HCC treatment,lenvatinib faces a significant hurdle due to the development of both intrinsic and acquired resistance among patients,and the underlying mechanism remains largely unknown.The present study aims to identify the pivotal gene responsible for lenvatinib resistance in HCC,explore the potential molecular mechanism,and propose combinatorial therapeutic targets for HCC management.Methods Cell viability and colony formation assays were conducted to evaluate the sensitivity of cells to lenvatinib and dicoumarol.RNA-Seq was used to determine the differences in transcriptome between parental cells and lenvatinib-resistant(LR)cells.The upregulated genes were analyzed by GO and KEGG analyses.Then,qPCR and Western blotting were employed to determine the relative gene expression levels.Afterwards,the intracellular reactive oxygen species(ROS)and apoptosis were detected by flow cytometry.Results PLC-LR and Hep3B-LR were established.There was a total of 116 significantly upregulated genes common to both LR cell lines.The GO and KEGG analyses indicated that these genes were involved in oxidoreductase and dehydrogenase activities,and reactive oxygen species pathways.Notably,NAD(P)H:quinone oxidoreductase 1(NQO1)was highly expressed in LR cells,and was involved in the lenvatinib resistance.The high expression of NQO1 decreased the production of ROS induced by lenvatinib,and subsequently suppressed the apoptosis.The combination of lenvatinib and NQO1 inhibitor,dicoumarol,reversed the resistance of LR cells.Conclusion The high NQO1 expression in HCC cells impedes the lenvatinib-induced apoptosis by regulating the ROS levels,thereby promoting lenvatinib resistance in HCC cells.展开更多
AIM: To investigate whether dicoumarol, a potent inhibitor of NAD(P)H quinone oxidoreductase-1 (NQO1), potentiates gemcitabine to induce cytotoxicity in chol-angiocarcinoma cells (CCA) and the role of reactive oxygen ...AIM: To investigate whether dicoumarol, a potent inhibitor of NAD(P)H quinone oxidoreductase-1 (NQO1), potentiates gemcitabine to induce cytotoxicity in chol-angiocarcinoma cells (CCA) and the role of reactive oxygen generation in sensitizing the cells. METHODS: Four human cell lines with different NQO1 activity were used; the human CCA cell lines, KKU-100, KKU-OCA17, KKU-M214, and Chang liver cells. NQO1 activity and mRNA expression were determined. The cells were pretreated with dicoumarol at relevant concentrations before treatment with gemcitabine. Cytotoxicity was determined by staining with fluorescent dyes. Oxidant formation was examined by assay of cellular glu-tathione levels and reactive oxygen species production by using dihydrofluorescein diacetate. Measurement of mitochondrial transmembrane potential was performed by using JC-1 fluorescent probe. Western blotting analysis was performed to determine levels of survival related proteins. RESULTS: Dicoumarol markedly enhanced the cytotoxicity of gemcitabine in KKU-100 and KKU-OCA17, the high NQO1 activity and mRNA expressing cells, but not in the other cells with low NQO1 activity. Dicoumarol induced a marked decrease in cellular redox of gluta-thione in KKU-100 cells, in contrast to KKU-M214 cells. Dicoumarol at concentrations that inhibited NQO1 activity did not alter mitochondrial transmembrane potential and production of reactive oxygen species. Gemcitabine alone induced activation of NF-κB and Bcl-XL protein expression. However, gemcitabine and dicoumarol combination induced increased p53 and decreased Bcl-XL levels in KKU-100, but not in KKU-M214 cells. CONCLUSION: NQO1 may be important in sensitizing cells to anticancer drugs and inhibition of NQO1 may be a strategy for the treatment of CCA.展开更多
Objective： To investigate the relationship between NAD（P）H：quinone oxidoreductase 1 （NQOI） C609T polymorphism and colon cancer risk in farmers from western region of Inner Mongolia. Methods： Polymerase chain re...Objective： To investigate the relationship between NAD（P）H：quinone oxidoreductase 1 （NQOI） C609T polymorphism and colon cancer risk in farmers from western region of Inner Mongolia. Methods： Polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） was performed to analyze NQO1 C609T polymorphism from 160 healthy controls and 76 colon cancer patients. Results： Among the colon cancer patients, the incidence of NQOI T allele （53.29%） was significantly higher than it in control group （33.75%, P〈0.001）. The individuals with NQO1 T allele had higher risk [2.239 （95% CI： 1.510-3.321） times] to develop colon cancer than individuals with NQO1 C allele. The incidence of NQO1 （TFI-） （34.21%） in colon cancer patients was higher than that in control group （15.62%, P〈0.001）. Odds ratios （OR） analysis suggested that NQO1 （T/F） and NQOI （T/C） genotype carriers had 3.813 （95% CI： 1.836-7.920） times and 2.080 （1.026-4.219） times risk compared with wild-type NQO1 （C/C） gene carriers in developing colon cancer. Individuals with NQO1 （T/I＇） genotype had 2.541 （95% CI： 0.990-6.552） times, 3.713 （95% CI： 1.542-8.935） times, and 3.471 （95% CI： 1.356-8.886） times risk than individuals with NQOI （T/C） or NQOI （C/C） genotype in well- differentiated, moderately-differentiated, and poorly-differentiated colon cancer patients, respectively. Conclusions： NQO1 gene C609T could be one of risk factors of colon cancer in farmers from western region of Inner Mongolia,展开更多
目的建立单管双向荧光PCR方法快速测定人NAD（P）H：醌氧化还原酶1[NAD（P）H：quinone oxidoreductase 1，NQ01]基因609C／T多态性。方法以人NQ01基因中的609C／T位点，设计双向引物，优化反应条件，应用SYBR Green Ⅰ双向荧光PCR扩增...目的建立单管双向荧光PCR方法快速测定人NAD（P）H：醌氧化还原酶1[NAD（P）H：quinone oxidoreductase 1，NQ01]基因609C／T多态性。方法以人NQ01基因中的609C／T位点，设计双向引物，优化反应条件，应用SYBR Green Ⅰ双向荧光PCR扩增191份人基因组DNA标本，并通过对产物进行熔解曲线分析，根据产物Tm值进行等位基因单核苷酸多态性分型。对其中62份标本用经典的PCR-限制性片段长度多态方法进行基因型分型，验证结果准确性。结果62份样本的单管双向荧光PCR方法的基因型结果与PCR-限制性片段长度多态法分型结果符合率100％。191份样本中，纯合野生型（CC）占28％，杂合型（CT）占50％，纯合突变型（TT）占22％。结论单管双向荧光PCR方法检测NQ01基因609C／T多态性，操作简便、反应过程快速，结果直观，敏感性、准确性和稳定性好，适用于临床样本检测及流行病学调查研究。展开更多
AIM:To determine whether the microRNA-27b-3p(miR-27b-3p)/NF-E2-related factor 2(Nrf2)pathway plays a role in human retinal pigment epithelial(hRPE)cell response to high glucose,how miR-27b-3p and Nrf2 expression are r...AIM:To determine whether the microRNA-27b-3p(miR-27b-3p)/NF-E2-related factor 2(Nrf2)pathway plays a role in human retinal pigment epithelial(hRPE)cell response to high glucose,how miR-27b-3p and Nrf2 expression are regulated,and whether this pathway could be specifically targeted.METHODS:hRPE cells were cultured in normal glucose or high glucose for 1,3,or 6d before measuring cellular proliferation rates using cell counting kit-8 and reactive oxygen species(ROS)levels using a dihydroethidium kit.miR-27b-3p,Nrf2,NAD(P)H quinone oxidoreductase 1(NQO1)and heme oxygenase-1(HO-1)mRNA and protein levels were analyzed using reverse transcription quantitative polymerase chain reaction(RT-qPCR)and immunocytofluorescence(ICF),respectively.Western blot analyses were performed to determine nuclear and total Nrf2 protein levels.Nrf2,NQO1,and HO-1 expression levels by RT-qPCR,ICF,or Western blot were further tested after miR-27b-3p overexpression or inhibitor lentiviral transfection.Finally,the expression level of those target genes was analyzed after treating hRPE cells with pyridoxamine.RESULTS:Persistent exposure to high glucose gradually suppressed hRPE Nrf2,NQO1,and HO-1 mRNA and protein levels and increased miR-27b-3p mRNA levels.High glucose also promoted ROS release and inhibited cellular proliferation.Nrf2,NQO1,and HO-1 mRNA levels decreased after miR-27b-3p overexpression and,conversely,both mRNA and protein levels increased after expressing a miR-27b-3p inhibitor.After treating hRPE cells exposed to high glucose with pyridoxamine,ROS levels tended to decreased,proliferation rate increased,Nrf2,NQO1,and HO-1 mRNA and protein levels were upregulated,and miR-27b-3p mRNA levels were suppressed.CONCLUSION:Nrf2 is a downstream target of miR-27b-3p.Furthermore,the miR-27b-3p inhibitor pyridoxamine can alleviate high glucose injury by regulating the miR-27b-3p/Nrf2 axis.展开更多
The two-electron cytoplasmic reductase NAD(P)H:quinone oxidoreductase 1 is expressed in many tissues.NAD(P)H:quinone oxidoreductase 1 is well-known for being highly expressed in most cancers.Therefore,it could be a ta...The two-electron cytoplasmic reductase NAD(P)H:quinone oxidoreductase 1 is expressed in many tissues.NAD(P)H:quinone oxidoreductase 1 is well-known for being highly expressed in most cancers.Therefore,it could be a target for cancer therapy.Because it is a quinone reductase,many bioimaging probes based on quinone structures target NAD(P)H:quinone oxidoreductase 1 to diagnose tumours.Its expression is higher in tumours than in normal tissues,and using target drugs such asβ-lapachone to reduce side effects in normal tissues can help.However,the physicochemical properties ofβlapachone limit its application.The problem can be solved by using nanosystems to deliverβ-lapachone.This minireview summarizes quinone-based fluorescent,nearinfrared and two-photon fluorescent probes,as well as nanosystems for delivering the NAD(P)H:quinone oxidoreductase 1-activating drugβ-lapachone.This review provides valuable information for the future development of probes and nano-delivery systems that target NAD(P)H:quinone oxidoreductase 1.展开更多
基金supported by the Global Select Project(No.DJK-LX-2022001)of the Institute of Health and Medicine,Hefei Comprehensive National Science Center.
文摘Objective Hepatocellular carcinoma(HCC)is the third leading cause of cancer-associated death worldwide.As a first-line drug for advanced HCC treatment,lenvatinib faces a significant hurdle due to the development of both intrinsic and acquired resistance among patients,and the underlying mechanism remains largely unknown.The present study aims to identify the pivotal gene responsible for lenvatinib resistance in HCC,explore the potential molecular mechanism,and propose combinatorial therapeutic targets for HCC management.Methods Cell viability and colony formation assays were conducted to evaluate the sensitivity of cells to lenvatinib and dicoumarol.RNA-Seq was used to determine the differences in transcriptome between parental cells and lenvatinib-resistant(LR)cells.The upregulated genes were analyzed by GO and KEGG analyses.Then,qPCR and Western blotting were employed to determine the relative gene expression levels.Afterwards,the intracellular reactive oxygen species(ROS)and apoptosis were detected by flow cytometry.Results PLC-LR and Hep3B-LR were established.There was a total of 116 significantly upregulated genes common to both LR cell lines.The GO and KEGG analyses indicated that these genes were involved in oxidoreductase and dehydrogenase activities,and reactive oxygen species pathways.Notably,NAD(P)H:quinone oxidoreductase 1(NQO1)was highly expressed in LR cells,and was involved in the lenvatinib resistance.The high expression of NQO1 decreased the production of ROS induced by lenvatinib,and subsequently suppressed the apoptosis.The combination of lenvatinib and NQO1 inhibitor,dicoumarol,reversed the resistance of LR cells.Conclusion The high NQO1 expression in HCC cells impedes the lenvatinib-induced apoptosis by regulating the ROS levels,thereby promoting lenvatinib resistance in HCC cells.
基金Supported by Thailand Research Fund, National Science and Technology Development Agency, research funding from KhonKaen Universitythe Royal Golden Jubilee Ph.D. Program (toKongpetch S)the Office of the Commission on Higher Educa-tion (to Buranrat B)
文摘AIM: To investigate whether dicoumarol, a potent inhibitor of NAD(P)H quinone oxidoreductase-1 (NQO1), potentiates gemcitabine to induce cytotoxicity in chol-angiocarcinoma cells (CCA) and the role of reactive oxygen generation in sensitizing the cells. METHODS: Four human cell lines with different NQO1 activity were used; the human CCA cell lines, KKU-100, KKU-OCA17, KKU-M214, and Chang liver cells. NQO1 activity and mRNA expression were determined. The cells were pretreated with dicoumarol at relevant concentrations before treatment with gemcitabine. Cytotoxicity was determined by staining with fluorescent dyes. Oxidant formation was examined by assay of cellular glu-tathione levels and reactive oxygen species production by using dihydrofluorescein diacetate. Measurement of mitochondrial transmembrane potential was performed by using JC-1 fluorescent probe. Western blotting analysis was performed to determine levels of survival related proteins. RESULTS: Dicoumarol markedly enhanced the cytotoxicity of gemcitabine in KKU-100 and KKU-OCA17, the high NQO1 activity and mRNA expressing cells, but not in the other cells with low NQO1 activity. Dicoumarol induced a marked decrease in cellular redox of gluta-thione in KKU-100 cells, in contrast to KKU-M214 cells. Dicoumarol at concentrations that inhibited NQO1 activity did not alter mitochondrial transmembrane potential and production of reactive oxygen species. Gemcitabine alone induced activation of NF-κB and Bcl-XL protein expression. However, gemcitabine and dicoumarol combination induced increased p53 and decreased Bcl-XL levels in KKU-100, but not in KKU-M214 cells. CONCLUSION: NQO1 may be important in sensitizing cells to anticancer drugs and inhibition of NQO1 may be a strategy for the treatment of CCA.
基金supported by the Autonomous region Natural Science Foundation of China,No. 200711020906
文摘Objective： To investigate the relationship between NAD（P）H：quinone oxidoreductase 1 （NQOI） C609T polymorphism and colon cancer risk in farmers from western region of Inner Mongolia. Methods： Polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） was performed to analyze NQO1 C609T polymorphism from 160 healthy controls and 76 colon cancer patients. Results： Among the colon cancer patients, the incidence of NQOI T allele （53.29%） was significantly higher than it in control group （33.75%, P〈0.001）. The individuals with NQO1 T allele had higher risk [2.239 （95% CI： 1.510-3.321） times] to develop colon cancer than individuals with NQO1 C allele. The incidence of NQO1 （TFI-） （34.21%） in colon cancer patients was higher than that in control group （15.62%, P〈0.001）. Odds ratios （OR） analysis suggested that NQO1 （T/F） and NQOI （T/C） genotype carriers had 3.813 （95% CI： 1.836-7.920） times and 2.080 （1.026-4.219） times risk compared with wild-type NQO1 （C/C） gene carriers in developing colon cancer. Individuals with NQO1 （T/I＇） genotype had 2.541 （95% CI： 0.990-6.552） times, 3.713 （95% CI： 1.542-8.935） times, and 3.471 （95% CI： 1.356-8.886） times risk than individuals with NQOI （T/C） or NQOI （C/C） genotype in well- differentiated, moderately-differentiated, and poorly-differentiated colon cancer patients, respectively. Conclusions： NQO1 gene C609T could be one of risk factors of colon cancer in farmers from western region of Inner Mongolia,
文摘目的建立单管双向荧光PCR方法快速测定人NAD（P）H：醌氧化还原酶1[NAD（P）H：quinone oxidoreductase 1，NQ01]基因609C／T多态性。方法以人NQ01基因中的609C／T位点，设计双向引物，优化反应条件，应用SYBR Green Ⅰ双向荧光PCR扩增191份人基因组DNA标本，并通过对产物进行熔解曲线分析，根据产物Tm值进行等位基因单核苷酸多态性分型。对其中62份标本用经典的PCR-限制性片段长度多态方法进行基因型分型，验证结果准确性。结果62份样本的单管双向荧光PCR方法的基因型结果与PCR-限制性片段长度多态法分型结果符合率100％。191份样本中，纯合野生型（CC）占28％，杂合型（CT）占50％，纯合突变型（TT）占22％。结论单管双向荧光PCR方法检测NQ01基因609C／T多态性，操作简便、反应过程快速，结果直观，敏感性、准确性和稳定性好，适用于临床样本检测及流行病学调查研究。
基金Supported by National Natural Science Foundation of China(No.2020J01652)the Training Project for Young and Middleaged Core Talents in Health System of Fujian Province(No.2016-ZQN-62).
文摘AIM:To determine whether the microRNA-27b-3p(miR-27b-3p)/NF-E2-related factor 2(Nrf2)pathway plays a role in human retinal pigment epithelial(hRPE)cell response to high glucose,how miR-27b-3p and Nrf2 expression are regulated,and whether this pathway could be specifically targeted.METHODS:hRPE cells were cultured in normal glucose or high glucose for 1,3,or 6d before measuring cellular proliferation rates using cell counting kit-8 and reactive oxygen species(ROS)levels using a dihydroethidium kit.miR-27b-3p,Nrf2,NAD(P)H quinone oxidoreductase 1(NQO1)and heme oxygenase-1(HO-1)mRNA and protein levels were analyzed using reverse transcription quantitative polymerase chain reaction(RT-qPCR)and immunocytofluorescence(ICF),respectively.Western blot analyses were performed to determine nuclear and total Nrf2 protein levels.Nrf2,NQO1,and HO-1 expression levels by RT-qPCR,ICF,or Western blot were further tested after miR-27b-3p overexpression or inhibitor lentiviral transfection.Finally,the expression level of those target genes was analyzed after treating hRPE cells with pyridoxamine.RESULTS:Persistent exposure to high glucose gradually suppressed hRPE Nrf2,NQO1,and HO-1 mRNA and protein levels and increased miR-27b-3p mRNA levels.High glucose also promoted ROS release and inhibited cellular proliferation.Nrf2,NQO1,and HO-1 mRNA levels decreased after miR-27b-3p overexpression and,conversely,both mRNA and protein levels increased after expressing a miR-27b-3p inhibitor.After treating hRPE cells exposed to high glucose with pyridoxamine,ROS levels tended to decreased,proliferation rate increased,Nrf2,NQO1,and HO-1 mRNA and protein levels were upregulated,and miR-27b-3p mRNA levels were suppressed.CONCLUSION:Nrf2 is a downstream target of miR-27b-3p.Furthermore,the miR-27b-3p inhibitor pyridoxamine can alleviate high glucose injury by regulating the miR-27b-3p/Nrf2 axis.
基金the financial support from the Tianjin Science and Technology Committee(Grant No.19JCYBJC28400)the Basic Research General Program of Shenzhen Science and Technology Innovation Commission in 2020(Grant No.JCYJ20190806162412752).
文摘The two-electron cytoplasmic reductase NAD(P)H:quinone oxidoreductase 1 is expressed in many tissues.NAD(P)H:quinone oxidoreductase 1 is well-known for being highly expressed in most cancers.Therefore,it could be a target for cancer therapy.Because it is a quinone reductase,many bioimaging probes based on quinone structures target NAD(P)H:quinone oxidoreductase 1 to diagnose tumours.Its expression is higher in tumours than in normal tissues,and using target drugs such asβ-lapachone to reduce side effects in normal tissues can help.However,the physicochemical properties ofβlapachone limit its application.The problem can be solved by using nanosystems to deliverβ-lapachone.This minireview summarizes quinone-based fluorescent,nearinfrared and two-photon fluorescent probes,as well as nanosystems for delivering the NAD(P)H:quinone oxidoreductase 1-activating drugβ-lapachone.This review provides valuable information for the future development of probes and nano-delivery systems that target NAD(P)H:quinone oxidoreductase 1.
