1.Development of fluorescence imaging-based system for screening compounds with antitumor activity.
Xiao-jing NIE ; Xiao-ping ZHAO ; Yi WANG
Journal of Zhejiang University. Medical sciences 2011;40(6):617-646
OBJECTIVETo develop a fluorescence imaging-based novel system for quick screening of antitumor compounds in vitro.
METHODSThe antitumor activity of 26 components from Lindera aggregate were determined by relative number of viable cell labelled with fluorescein diacetate (FDA) in multiwell plates after exposure to these 26 different components. Then, the linearity and precision of this method were validated. The structures of active compounds in components with strong antitumor activity were deduced by LC/MS.
RESULTSThe linearity of this method for cells stained with FDA was validated (r² = 0.9858) in the range of 0-10⁴ cells per well, and the in-plate precision was 9.41 %. Two of 26 components from Lindera aggregate showed significant inhibition effect on proliferation of HepG2 cells (inhibition rate >90%).
CONCLUSIONThis proposed rapid and reliable approach can be used for screening compounds with antitumor activity from Traditional Chinese Medicine in vitro. The major active compound of Lindera aggregate was putatively identified as norboldine by LC/MS analysis.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Drug Screening Assays, Antitumor ; methods ; Fluorescence ; Humans
2.Establishing the method of collagen-gel droplet embedded culture drug sensitivity test with rat tail collagen.
Ya HU ; Tao ZHOU ; Quan LIAO ; Tai-Ping ZHANG ; Ge CHEN ; Yu-Pei ZHAO
Chinese Journal of Surgery 2008;46(13):998-1001
OBJECTIVETo explore the clinical application of collagen-gel droplet embedded-culture drug sensitivity test (CD-DST) in the malignant tumors.
METHODSCD-DST was established with rat tail collagen. Three pancreatic cancer cell lines, surgical resection specimens including 15 cases of pancreatic cancer and 10 cases of gastrointestinal cancer were examined using CD-DST.
RESULTSThe overall achievement ratio of CD-DST for clinical tumor specimens was 80% (20/25). In vitro chemosensitivities of pancreatic carcinoma cells to 5-FU, gemcitabine and oxaliplatin were lower than those of gastrointestinal carcinoma.
CONCLUSIONSCD-DST with rat tail collagen is a valuable chemosensitivity testing method for malignant tumors. It can be used to realize individualized anticancer chemotherapy.
Animals ; Collagen ; Drug Screening Assays, Antitumor ; methods ; Female ; Gels ; Humans ; Male ; Rats ; Tail ; chemistry ; Tumor Cells, Cultured
3.A method of screening the antitumor lead compounds based on the dynamic bio-response profile of cells.
Li-Na MA ; Le-Le ZHANG ; Yin XIONG ; Yu-Mei HAN ; Cong-En ZHANG ; Dan GAO ; Li MA ; Dan YAN ; Xiao-He XIAO
Acta Pharmaceutica Sinica 2014;49(5):695-700
The study is to report the establishment of a method of screening the antitumor compounds based on the dynamic bio-response profile of cells to make up for the shortages of conventional end-point tests such as tedious operation and low sensitivity. Based on the principle of electric impedance of cells, the real-time cell electronic sensing (RT-CES) system was used to monitor the effect of epirubicin (EPI), cisplatinum (DDP) and carboplatin (CBP) on the growth of HepG2 cells, with the cell index (CI), half maximal inhibitory concentration (IC50) and detachment curve as evaluation indexes. Meanwhile, cell counting kit-8 (CCK-8) and microscopy were applied for verification. The results showed that CI curve could sensitively real-time profile the inhibitory effect of model drugs on HepG2 cells. The IC50 of EPI, DDP and CBP were 0.53 +/- 0.04, 9.79 +/- 0.26 and 597.00 +/- 3.79 microg x mL(-1), respectively. What's more, the significant differences of detachment curves of the three drugs indicated that their functional mechanisms might be different, this is consistent with the literature. The RT-CES system with non-invasive, label-free and real-time characteristics could be used to monitor the bio-response profile of the three drugs to HepG2 cells, allowing to qualitatively and quantitatively distinguish the antitumor activities of the three drugs, and could be a complementary method for the present screening of antitumor compounds.
Antineoplastic Agents
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pharmacology
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Biosensing Techniques
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methods
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Cell Count
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Cell Line, Tumor
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Cisplatin
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pharmacology
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Drug Screening Assays, Antitumor
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Electric Impedance
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Humans
4.Application of ATP based bioluminescence tumor chemosensitivity assay in the chemotherapy of pediatric solid tumor.
Hai-yan CHENG ; Xiu-dan ZHU ; Huan-min WANG ; Hong QIN
Chinese Journal of Pediatrics 2009;47(8):598-603
OBJECTIVETo explore the clinical significance of ATP based bioluminescence in vitro tumor chemosensitivity assay (ATP-TCA) in the chemotherapy of pediatric solid tumor.
METHODSThe cell culture technique and ATP-TCA were used to study chemosensitivity assay in specimens from 50 cases who underwent resection surgery for solid tumor (15 malignant neurogenic tumor, 8 malignant germ cell tumors, 10 Wilms' tumors, 10 hepatoblastomas, 6 rhabdomyosarcomas, 1 adrenocortical carcinoma), 8 chemotherapeutic drugs and 8 drug combination schedules were applied in every specimen.
RESULTS(1) Specimens of 46 of 50 pediatric patients with solid tumors were suitable for evaluation and were evaluated, the overall evaluation rate was 92% (46/50). (2) There was the heterogeneity in the chemosensitivity of the solid tumors in vitro. (3) The drug combination schedules of high sensitivity rate of every kind of pediatric solid tumor are as follows: the malignant neurogenic tumor: CBP + EPI + IFO (12/15, 80.0%), VCR + CTX + DDP + DTIC (11/15, 73.3%); malignant germ cell tumor: DDP + VCR + BLM(8/8, 100%), TPTN + ACTD + IFO(8/8, 100%), As2O3 (7/8, 87.5%); Wilms' tumor: VCR + ACTD(6/7, 85.7%), CBP + VP16 (6/8, 75.0%); hepatoblastoma: VCR + CTX + DDP + VP16 (8/9, 88.9%), CBP +IFO + VM26 (7/9, 77.8%), DDP + VP16 + TPTN(7/9, 77.8%); rhabdomyosarcoma: VCR + CTX + DDP + VP16 (5/5, 100%); adrenocortical carcinoma: VCR + CTX + ADM. (4) As2O3 reached a high in vitro sensitive rate of 87.5% (7/8) and 46.7% (7/15) in malignant germ cell tumor and the malignant neurogenic tumor respectively, PTX was sensitive to the malignant neurogenic tumor and rhabdomyosarcoma (40.0% (6/15), 60.0% (3/5)), GEM was sensitive to pediatric malignant germ cell tumor and rhabdomyosarcoma (50.0% (4/8), 60.0% (3/5)).
CONCLUSIONSATP-TCA is a sensitive method for the chemotherapeutic agents screening of pediatric malignant solid tumor, and ATP-TCA assay results correlated well with clinical response. It appears to be useful in screening new drugs for pediatric solid tumor, exploring the possible combination plots and principles, evaluating the efficacy of existing chemotherapy, and optimize chemotherapy on an individual basis.
Adolescent ; Antineoplastic Agents ; Child ; Child, Preschool ; Drug Screening Assays, Antitumor ; methods ; Female ; Humans ; In Vitro Techniques ; Infant ; Male
5.An automatic fluorescence chemosensitivity analysis system for tumors.
Chinese Journal of Medical Instrumentation 2005;29(3):167-169
The Automatic Fluorescence Chemosensitivity Analysis System (AFCAS), on the basis of image analysis, image recognition and database technologies, automatically collects the microscopic fluorescent images, inputs patient information, processes images, counts cell number, analyzes statistical data, saves and screens the reports and thus it is able to provide effective informations for assisting medical diagnosis and treatments of tumors or cancers. The AFCAS System adopts the image recognition technology of Region Chain Code and Bayesian Classification and ADO-Based database technology.
Algorithms
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Computers
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Drug Screening Assays, Antitumor
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methods
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Equipment Design
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Image Processing, Computer-Assisted
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methods
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Information Storage and Retrieval
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methods
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Pattern Recognition, Automated
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methods
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Software
7.Effect of several anti-tumor drugs on apoptosis induction in Jurkat cell line.
Yong-Qiu MAO ; Xi-Rong LI ; Song LEI
Journal of Experimental Hematology 2006;14(4):681-685
Cisplatin (CDDP), homoharringtonine (HHT), mitoxantrone (MIT) and hydroxycamptothecin (HCPT) are highly effective anti-tumor drugs. To evaluate their effects in the therapy of leukemia and establish a valuable method to estimate anti-tumor drugs, Annexin V/PI double parameter flow cytometry was used to detect the effects of these drug inducing apoptosis and death in Jurkat cell line. The results showed that MIT and HCTP-induced apoptosis effects on Jurkat cell line were obvious at 4 hours in early phase after adding drug (P < 0.05) and at 8 hours in late phase after adding drug (P < 0.05). HHT had obvious effect on inducing apoptosis of Jurkat cells, but no significant difference from low to high doses. The effect of CDDP on inducing apoptosis of Jurkat cell line was obviously weaker than that of HHT, MIT and HCPT, its weak effect on apoptosis of Jurkat cell line was found only at high concentration of drug for long time. Death effects on Jurkat cell line can not be observed in every experimental group. It is concluded that low dose of MIT can effectively induced apoptosis of Jurkat cell line. Annexin V/PI double parameter flow cytometry can be used as a reliable method for clinical screening anti-tumor drugs.
Antineoplastic Agents
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pharmacology
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Apoptosis
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drug effects
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Camptothecin
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pharmacology
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Cisplatin
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pharmacology
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Drug Screening Assays, Antitumor
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methods
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Harringtonines
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pharmacology
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Humans
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Jurkat Cells
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Mitoxantrone
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pharmacology
8.Enhancing the cytotoxicity of antitumor drugs through electromagnetic pulses.
Guo YANG ; Baoyi WANG ; Hong ZHANG ; Haichuan CHEN ; Zishu WANG ; Kong YANG ; Jingru SUN
Journal of Biomedical Engineering 2003;20(3):497-499
We chose Hela cells as research object and studied the cytotoxicity generated by cyclophosphamide, an antitumor drug, after cell electroporation by the use of electromagnetic pulses. Comparison between the electroporation group and the contrast group revealed the greatly enhanced cytotoxicity of the electroporation group, indicating that under some conditions electromagnetic pulses can enhance the cytotoxicity of antitumor drugs. The results of this study provide reliable evidences and a feasible approach for clinical treatment of tumor.
Antineoplastic Agents
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pharmacology
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Apoptosis
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Cell Survival
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drug effects
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Cyclophosphamide
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pharmacology
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Drug Screening Assays, Antitumor
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Electromagnetic Phenomena
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Electroporation
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methods
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HeLa Cells
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Humans
9.Application of temperature sensitive yeast models with definite target in the screening of potential human Pin1 inhibitors.
Jing ZHANG ; Xiao-Min HAN ; Wen-Hui HU ; Zong-Ru GUO ; Xiao-Bo HE ; Shu-Yi SI
Acta Pharmaceutica Sinica 2014;49(6):854-860
This study is to explore new lead compounds by inhibition of Pin1 for anticancer therapy using temperature sensitive mutants. As Pin1 is conserved from yeast to human, we established a high-throughput screening method for Pin1 inhibitors, which employed yeast assay. This method led to the identification of one potent hits, 8-11. In vitro, 8-11 inhibited purified Pin1 enzyme activity with IC50 of (10.40 +/- 1.68) micromol x L(-1), induced G1 phase arrest and apoptosis, showed inhibitory effects on a series of cancer cell proliferation, reduced Cyclin D1 expression, was defined as reciprocally matched for protein-ligand complex in virtual docking analysis and reduced cell migration ability. In vivo, we could observe reduction of tumor volume after treatment with 8-11 in xenograft mice compared with vehicle DMSO treatment. Altogether, these results provide for the first time the involvement of 8-11 in the anticancer activity against Pin1.
Animals
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Apoptosis
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drug effects
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Cell Proliferation
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drug effects
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Cyclin D1
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metabolism
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Drug Screening Assays, Antitumor
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methods
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G1 Phase
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High-Throughput Screening Assays
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methods
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Humans
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Mice
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NIMA-Interacting Peptidylprolyl Isomerase
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Neoplasms
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pathology
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Peptidylprolyl Isomerase
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antagonists & inhibitors
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Temperature
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Xenograft Model Antitumor Assays
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Yeasts
10.MTT assay for detecting 5-fluorouracil chemosensitivity of human breast carcinoma cell line.
Fang-xuan HAN ; Hui LIN ; Ling RU
Journal of Southern Medical University 2009;29(1):97-99
OBJECTIVETo assess the feasibility of MTT colorimetric assay for testing the in vitro chemosensitivity of breast cancer cells to 5 fluorouracil (5-Fu).
METHODSThe chemosensitivity of human breast carcinoma cell lines MCF-7 and MDA-MB-435S to 5-Fu at different concentrations was evaluated with MTT assay.
RESULTS5-FU treatment resulted in dose-dependent growth inhibition of the breast cancer cells with both low and high metastatic capacities.
CONCLUSIONSMTT assay may help select appropriate chemotherapeutic agents and provides evidence for individualized chemotherapy for breast cancer.
Antimetabolites, Antineoplastic ; pharmacology ; Breast Neoplasms ; pathology ; Cell Line, Tumor ; Colorimetry ; methods ; Coloring Agents ; Drug Screening Assays, Antitumor ; Female ; Fluorouracil ; pharmacology ; Humans ; Tetrazolium Salts