Natural products include several diverse compounds that have been found to be effective against cancer.Discovering anticancer compounds in nature is a multistep and complex process that requires pre-clinical and clini...Natural products include several diverse compounds that have been found to be effective against cancer.Discovering anticancer compounds in nature is a multistep and complex process that requires pre-clinical and clinical studies.Only a few of the available natural products are used to treat cancer since most of them have very high complexity and low bioavailability.Therefore,the process of anticancer drug discovery requires a straightforward and effective method to assess anticancer activity using in vitro assays.This review summarizes various cell-based assays and techniques used to measure cell viability,migration,and apoptosis,focusing in particular on the principles,mechanisms,advantages,and disadvantages of each assay to provide a preliminary platform for cancer drug discovery.展开更多
Traditional tumor models do not tend to accurately simulate tumor growth in vitro or enable personalized treatment and are particularly unable to discover more beneficial targeted drugs.To address this,this study desc...Traditional tumor models do not tend to accurately simulate tumor growth in vitro or enable personalized treatment and are particularly unable to discover more beneficial targeted drugs.To address this,this study describes the use of threedimensional(3D)bioprinting technology to construct a 3D model with human hepatocarcinoma SMMC-7721 cells(3DP-7721)by combining gelatin methacrylate(GelMA)and poly(ethylene oxide)(PEO)as two immiscible aqueous phases to form a bioink and innovatively applying fluorescent carbon quantum dots for long-term tracking of cells.The GelMA(10%,mass fraction)and PEO(1.6%,mass fraction)hydrogel with 3:1 volume ratio offered distinct pore-forming characteristics,satisfactorymechanical properties,and biocompatibility for the creation of the 3DP-7721 model.Immunofluorescence analysis and quantitative real-time fluorescence polymerase chain reaction(PCR)were used to evaluate the biological properties of the model.Compared with the two-dimensional culture cell model(2D-7721)and the 3D mixed culture cell model(3DM-7721),3DP-7721 significantly improved the proliferation of cells and expression of tumor-related proteins and genes.Moreover,we evaluated the differences between the three culture models and the effectiveness of antitumor drugs in the three models and discovered that the efficacy of antitumor drugs varied because of significant differences in resistance proteins and genes between the three models.In addition,the comparison of tumor formation in the three models found that the cells cultured by the 3DP-7721 model had strong tumorigenicity in nude mice.Immunohistochemical evaluation of the levels of biochemical indicators related to the formation of solid tumors showed that the 3DP-7721 model group exhibited pathological characteristics of malignant tumors,the generated solid tumors were similar to actual tumors,and the deterioration was higher.This research therefore acts as a foundation for the application of 3DP-7721 models in drug development research.展开更多
Making use of the fact that the combination of a drug substance with DNA may inhibit the duplication, synthesis and proliferation of DNA and the consistency of the in vivo and in vitro interactions, the authors worked...Making use of the fact that the combination of a drug substance with DNA may inhibit the duplication, synthesis and proliferation of DNA and the consistency of the in vivo and in vitro interactions, the authors worked out a preliminary screening method for testing complex agents as potential antitumor drugs using ethidium bromide as a fluorescence probe. In this report, the method was applied for in vitro testing fourteen synthesized palladium(Ⅱ)/phenanthroline/amino acid/chloride complexes as potential non-platinum antitumor agents. The fluorimetric screening method was compared with methylene blue tube test and trypan blue dye exclusion assay. All three methods gave agreeable results. Among the complexes tested, [Pd(phen)(lys)]Cl, [Pd(phen)(arg)]Cl and [Pd(phen)(pro)]Cl showed antineoplastic ratios for animal tumor S-180 56%, 50% and 48%, respectively, in accordance with the order of their binding constants with DNA, 7.96×10~6, 4.52×10~6 and 1.0×10~6, respectively. The test results show that fluorimetric method is simple, cheap and rapid. suitable for preliminary screening of antitumor complexes.展开更多
Current in vitro assays for the activity of HIV-RT(reverse transcriptase)require radio-labeled or chemically modified nucleotides to detect reaction products.However,these assays are inherently end-point measurements ...Current in vitro assays for the activity of HIV-RT(reverse transcriptase)require radio-labeled or chemically modified nucleotides to detect reaction products.However,these assays are inherently end-point measurements and labor intensive.Here we describe a novel non-radioactive assay based on the principle of pyrosequencing coupledenzyme system to monitor the activity of HIV-RT by indirectly measuring the release of pyrophosphate(PPi),which is generated during nascent strand synthesis.The results show that our assay could monitor HIV-RT activity with high sensitivity and is suitable for rapid highthroughput drug screening targeting anti-HIV therapies due to its high speed and convenience.Moreover,this assay can be used to measure primase activity in an easy and sensitive manner,which suggests that this novel approach could be wildly used to analyze the activity of PPi-generated and ATP-free enzyme reactions.展开更多
目的:本研究旨在探讨肿瘤特殊转移灶恶性胸腔/腹腔积液中核苷酸切除修复交叉互补组1(excision repair cross-complementing group 1,ERCC1)基因和乳腺癌易感基因1(breast cancer 1,BRCA1)的表达水平与顺铂体外药物敏感性之间的关系。方...目的:本研究旨在探讨肿瘤特殊转移灶恶性胸腔/腹腔积液中核苷酸切除修复交叉互补组1(excision repair cross-complementing group 1,ERCC1)基因和乳腺癌易感基因1(breast cancer 1,BRCA1)的表达水平与顺铂体外药物敏感性之间的关系。方法:前瞻性收集46例Ⅳ期恶性肿瘤患者的恶性胸腔/腹腔积液,分离出肿瘤细胞后,采用体外药敏试验三磷酸腺苷生物荧光法检测(adenosine triphosphate-bioluminescence assay,ATP-TCA)法检测肿瘤细胞对顺铂的药物敏感性,实时荧光定量PCR检测ERCC1和BRCA1 mRNA的相对表达水平。结果:在非小细胞肺癌患者的恶性胸腔/腹腔积液中,肿瘤细胞的ERCC1 mRNA相对表达水平与顺铂抗药性呈正相关(P=0.001,r=0.685)。非小细胞肺癌患者(P=0.014,r=0.541)和胃癌患者(P=0.002,r=0.625)恶性胸腔/腹腔积液中肿瘤细胞的BRCA 1 mRNA相对表达水平与顺铂抗药性呈正相关。ERCC1和BRCA1对顺铂药物敏感性的影响存在交互作用(所有患者P=0.010;胃癌患者P=0.027)。结论:恶性胸腔/腹腔积液中肿瘤细胞的ERCC1和BRCA1 mRNA的相对表达水平与顺铂体外药物敏感性相关。联合检测ERCC1和BRCA1这2种基因较只采用其中一种基因在预测顺铂药物敏感性方面的价值更大。展开更多
基金supported by the Internal Research Grant of Sanata Dharma University No.007/Penel./LPPM-USD/II/2022.
文摘Natural products include several diverse compounds that have been found to be effective against cancer.Discovering anticancer compounds in nature is a multistep and complex process that requires pre-clinical and clinical studies.Only a few of the available natural products are used to treat cancer since most of them have very high complexity and low bioavailability.Therefore,the process of anticancer drug discovery requires a straightforward and effective method to assess anticancer activity using in vitro assays.This review summarizes various cell-based assays and techniques used to measure cell viability,migration,and apoptosis,focusing in particular on the principles,mechanisms,advantages,and disadvantages of each assay to provide a preliminary platform for cancer drug discovery.
基金supported by the National Natural Science Foundation of China(Nos.51975400 and 62031022)Shanxi Provincial Key Medical Scientific Research Project(Nos.2020XM06 and 2021XM12)+3 种基金Fundamental Research Program of Shanxi Province(No.202103021224081)Shanxi Provincial Basic Research Project(Nos.202103021221006 and 202103021223040)Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi(No.2021L044)Shanxi-Zheda Institute of Advanced Materials and Chemical Engineering(No.2022SX-TD026).
文摘Traditional tumor models do not tend to accurately simulate tumor growth in vitro or enable personalized treatment and are particularly unable to discover more beneficial targeted drugs.To address this,this study describes the use of threedimensional(3D)bioprinting technology to construct a 3D model with human hepatocarcinoma SMMC-7721 cells(3DP-7721)by combining gelatin methacrylate(GelMA)and poly(ethylene oxide)(PEO)as two immiscible aqueous phases to form a bioink and innovatively applying fluorescent carbon quantum dots for long-term tracking of cells.The GelMA(10%,mass fraction)and PEO(1.6%,mass fraction)hydrogel with 3:1 volume ratio offered distinct pore-forming characteristics,satisfactorymechanical properties,and biocompatibility for the creation of the 3DP-7721 model.Immunofluorescence analysis and quantitative real-time fluorescence polymerase chain reaction(PCR)were used to evaluate the biological properties of the model.Compared with the two-dimensional culture cell model(2D-7721)and the 3D mixed culture cell model(3DM-7721),3DP-7721 significantly improved the proliferation of cells and expression of tumor-related proteins and genes.Moreover,we evaluated the differences between the three culture models and the effectiveness of antitumor drugs in the three models and discovered that the efficacy of antitumor drugs varied because of significant differences in resistance proteins and genes between the three models.In addition,the comparison of tumor formation in the three models found that the cells cultured by the 3DP-7721 model had strong tumorigenicity in nude mice.Immunohistochemical evaluation of the levels of biochemical indicators related to the formation of solid tumors showed that the 3DP-7721 model group exhibited pathological characteristics of malignant tumors,the generated solid tumors were similar to actual tumors,and the deterioration was higher.This research therefore acts as a foundation for the application of 3DP-7721 models in drug development research.
基金Project supported by the National Education Commission Foundation and the National Natural Science Foundation of China.
文摘Making use of the fact that the combination of a drug substance with DNA may inhibit the duplication, synthesis and proliferation of DNA and the consistency of the in vivo and in vitro interactions, the authors worked out a preliminary screening method for testing complex agents as potential antitumor drugs using ethidium bromide as a fluorescence probe. In this report, the method was applied for in vitro testing fourteen synthesized palladium(Ⅱ)/phenanthroline/amino acid/chloride complexes as potential non-platinum antitumor agents. The fluorimetric screening method was compared with methylene blue tube test and trypan blue dye exclusion assay. All three methods gave agreeable results. Among the complexes tested, [Pd(phen)(lys)]Cl, [Pd(phen)(arg)]Cl and [Pd(phen)(pro)]Cl showed antineoplastic ratios for animal tumor S-180 56%, 50% and 48%, respectively, in accordance with the order of their binding constants with DNA, 7.96×10~6, 4.52×10~6 and 1.0×10~6, respectively. The test results show that fluorimetric method is simple, cheap and rapid. suitable for preliminary screening of antitumor complexes.
基金the National Natural Science Foundation of China(Grant Nos.30221003,30720022)the Ministry of Science and Technology 973 Project(Grant No.2006CB806503)+2 种基金the Ministry of Science and Technology National High Technology Research and Development Program("863"Program)(Grant No.2006AA02A322)the Ministry of Science and Technology International Cooperation Project(Grant No.2006DFB32420)the Chinese Academy of Sciences Knowledge Innovation Project(Grant No.KSCX1-YW-R-05)。
文摘Current in vitro assays for the activity of HIV-RT(reverse transcriptase)require radio-labeled or chemically modified nucleotides to detect reaction products.However,these assays are inherently end-point measurements and labor intensive.Here we describe a novel non-radioactive assay based on the principle of pyrosequencing coupledenzyme system to monitor the activity of HIV-RT by indirectly measuring the release of pyrophosphate(PPi),which is generated during nascent strand synthesis.The results show that our assay could monitor HIV-RT activity with high sensitivity and is suitable for rapid highthroughput drug screening targeting anti-HIV therapies due to its high speed and convenience.Moreover,this assay can be used to measure primase activity in an easy and sensitive manner,which suggests that this novel approach could be wildly used to analyze the activity of PPi-generated and ATP-free enzyme reactions.
