1.Effect of Multidrug Resistance Gene-1 (mdr1) Overexpression on In-Vitro Uptake of 99mTc-sestaMIBI in Murine L1210 Leukemia Cells.
Kyu Bo LEE ; June Key CHUNG ; Sang Kyun SOHN ; Kyung Ah CHUN ; Do Young KANG ; Sang Woo LEE ; Jaetae LEE ; Jong Kee LEE ; Soo Han JUN
Korean Journal of Nuclear Medicine 1999;33(2):152-162
PURPOSE: To determine whether Tc-99mMIBI is recognized by the multidrug resistant P-glycoprotein (Pgp), we have measured quantitatively Tc-99mMIBI uptake in cancer cells. The effects of various Pgp reversing agents on cellular Tc-99m-MIBI uptake were also investigated in the presence of multidrug resistance gene-1 (mdr1 gene) overexpression. MATERIALS AND METHODS: We measured percentage uptake of Tc-99m-MIBI at different incubation temperatures both in mdr1 positive and negative cells. The effects of verapamil, cyclosporin, and dipyridamole on cellular uptake of Tc-99m-MIBI were also evaluated with or without overexpression of mdr1 gene in cultured murine leukemia L1210 cells. RESULTS: The mdr1 gene expressing cell lines were effectively induced in in vitro with continuous application of low-dose adriamycin or vincristine. Cellular uptake of Tc-99m-MIBI was higher in mdr1 negative L1210 cells than those of mdr1 positive cells, and higher when incubated in 37 degree C than 4 degree C. In the presence of verapamil, cyclosporin or dipyridamole, Tc-99m-MIBI uptake was increased upto 604% in mdr1 positive cells. CONCLUSION: Cellular uptake of Tc-99m-MIBI is lower in leukemia cells over-expressing mdr1 gene, and MDR-reversing agents increase cellular uptake. These results suggest that Tc-99m-MIBI can be used for characterizing Pgp expression and developing MDR-reversing agents In vitro.
Animals
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Cell Line
;
Cyclosporine
;
Dipyridamole
;
Doxorubicin
;
Drug Resistance, Multiple*
;
Leukemia L1210
;
Leukemia*
;
P-Glycoprotein
;
Technetium Tc 99m Sestamibi*
;
Verapamil
;
Vincristine
2.Cloning of adriamycin-resistant related (arr) gene in an adriamycin-resistant L1210 variant.
Jae Ryong KIM ; Seong Yong KIM ; Jung Hye KIM
Experimental & Molecular Medicine 1998;30(3):145-149
A partial fragment of novel sequence (arr, adriamycin-resistant related) was previously identified using the differential display (DD)-PCR technique with adriamycin-resistant L1210 variant (L1210AdR), which shows a typical multidrug resistant (MDR) phenotypes. The present research shows the isolation of full length arr cDNA sequence. To clone the full length cDNA of arr gene, DD-PCR fragments were subjected to 5'- and 3'-Rapid Amplification of cDNA End (RACE) method. The cloned arr cDNA consisted of 770 bases and contained an open reading frame of 153 bases, encoding a protein of 51 amino acid with the molecular mass of 4 kDa by in vitro translation reactions. Northern blot analysis showed that a 770 bases transcript arr gene was overexpressed in adriamycin-resistant L1210 variant, but not in the parent suggesting that the arr gene may be involved in the adriamycin-resistant phenotypes.
Amino Acid Sequence
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Animal
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Base Sequence
;
Blotting, Northern
;
Cloning, Molecular
;
DNA Primers
;
Doxorubicin/pharmacology*
;
Drug Resistance, Neoplasm/genetics*
;
Gene Expression Regulation, Neoplastic
;
Glycoproteins/genetics*
;
Leukemia L1210/genetics*
;
Leukemia L1210/drug therapy
;
Mice
;
Molecular Sequence Data
;
Translation, Genetic
3.The effect of extremely low frequency magnetic fields on cytochrome oxidase subunit 1 mRNA transcription.
Tao ZHONG ; Qing CHEN ; Ruiying WU ; Gengdong YAO ; Deqiang LU ; Huai CHIANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2002;20(4):249-251
OBJECTIVETo clone and identify MF-1 gene which responded to extremely low frequency magnetic fields(ELF MF) in Daudi cells, and explore the response universality of MF-1 gene in several MF-sensitive cell lines, so as to provide experimental basis for revealing the mechanism of biological effects induced by magnetic field.
METHODSThe DNA fragment of MF-1 was cloned and sequenced; the mRNA level of MF-1 gene were analysed in MF-sensitive cell lines(HL-60, L1210 and CHL) by Northern blot after these cells being treated with 0.1 mT and 0.8 mT MF for 20 minutes and 24 hours, respectively.
RESULTSThe MF-1 cDNA sequence had 100% homology with cytochrome oxidase subunit 1 gene(CO1) by searching Gene Bank database; the transcription of CO1 in HL-60, L1210 and CHL cell lines which exposed to 0.1 mT and 0.8 mT MF for 20 minutes were significantly lower(0.38 +/- 0.12 and 0.37 +/- 0.04) than that of control(0.58 +/- 0.12) and so did for 24 hours exposure(0.46 +/- 0.09 and 0.45 +/- 0.09 vs 0.65 +/- 0.06) (P < 0.05).
CONCLUSIONCO1 is a MF-responsive gene. Cytochrome oxidase activity may be affected through low level of CO1 transcription by magnetic fields, thus induce bioeffects in organisms.
Animals ; Cricetinae ; Electron Transport Complex IV ; genetics ; metabolism ; radiation effects ; HL-60 Cells ; Humans ; Leukemia L1210 ; Magnetics ; Mice ; Protein Subunits ; RNA, Messenger ; analysis ; Transcription, Genetic ; radiation effects
4.Synthesis of methotrexate-poly (ethylene glycol) conjugate and their anti-tumor activity in vitro.
Jian-Feng ZHANG ; Dong-Zhi WEI ; Xiong ZHOU ; Feng JIANG
Acta Pharmaceutica Sinica 2007;42(6):607-610
To improve the physical property and bioactivity of methotrexate, this paper investigated the new formation of conjugate methotrexate-poly (ethylene glycol) and in vitro anti-tumor activity of the synthesized conjugate. The conjugate of methotrexate-poly (ethylene glycol), which was verified by the spectroscopy analysis of UV, IR and 13C NMR, was synthesized by chemical catalysis and micro-wave irritation. The determination for the conjugate of solubility in water and distribution coefficient in octanol-water system of the conjugate was done to examine its deliquescence property. The solubility in water and the distribution coefficient of the conjugate was greatly improved, which was increased by 128 folds and 5 folds, respectively. The in vitro anti-tumor activity of the conjugate was tested by mouse L(1210) leukaemia cells, and the synthesized conjugate showed the same anti-tumor activity as the original methotrexate. Compared to the reported literature, the modification of methotrexate by poly (ethylene glycol) is more rapid and convenient.
Animals
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Antineoplastic Agents
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chemical synthesis
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Leukemia L1210
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drug therapy
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Methotrexate
;
chemical synthesis
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chemistry
;
pharmacology
;
Mice
;
Polyethylene Glycols
;
chemical synthesis
;
chemistry
;
pharmacology
;
Solubility
5.Regulatory effect of resveratrol on JAK1/STAT3 signal transduction pathway in leukemia.
Journal of Experimental Hematology 2008;16(4):772-776
The aim of this study was to explore the molecule mechanism of resveratrol antileukaemia. The mouse lymphocytic leukemia L1210 cells were cultured and the expressions of pJAK1 and pSTAT3 protein in L1210 cells were detected by immunohistochemistry and immunoprecipitation in vitro. The mouse model with L1210 leukemia ascites carcinoma was established and activities of singal transduction pathway molecules pJAK1 and pSTAT3 were measured by Western blot and immunohistochemistry assay in vitro. The results indicated that resveratrol could significantly inhibit the JAK1/STAT3 signal transduction pathway, down-regulate expressions of pJAK1 and pSTAT3 and reduce the phosphorylation of JAK1 and STAT3 in a dose-and time-dependent manner. It is concluded that the resveratrol can regulate signal transduction pathway and reduce the activation of JAK1/STAT3 tyrosine phosphorylation significantly, and therefore resveratrol shows chemotherapeutic potential to leukaemia.
Animals
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Antineoplastic Agents, Phytogenic
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pharmacology
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Down-Regulation
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Janus Kinase 1
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genetics
;
metabolism
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Leukemia L1210
;
genetics
;
metabolism
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Mice
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Mice, Inbred BALB C
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Random Allocation
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STAT3 Transcription Factor
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genetics
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metabolism
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Signal Transduction
;
drug effects
;
Stilbenes
;
pharmacology
6.Synthesis and antitumor activity of anthracene-9-carbaldehyde amino-s-triazole Schiff-bases with side-chain of S-acetic acid.
Guo-Qiang HU ; Xiu-Li DONG ; Song-Qiang XIE ; Wen-Long HUANG
Acta Pharmaceutica Sinica 2008;43(1):50-53
To find out a novel lead compound from heterocyclic amine Schiff bases for developing new antitumor agents, each of (4-amino-5-substituted-s-triazol-3-ylthio) -acetic acids 2a-j was condensed with anthracene-9-carbaldehyde to obtain Schiff-bases of [4-(anthracen-9-yl methylene) amino] -5-substituted-s-triazol-3-ylsulfanyl] -acetic acids 3a-j. The structures of new compounds synthesized were characterized by elemental analysis and spectral data, and in vitro antitumor activity was also evaluated against CHO, HL60 and L1210 cell lines by MTT assay.
Acetic Acid
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chemistry
;
Animals
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Anthracenes
;
chemistry
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Antineoplastic Agents
;
chemical synthesis
;
chemistry
;
pharmacology
;
CHO Cells
;
Cricetinae
;
Cricetulus
;
HL-60 Cells
;
Humans
;
Leukemia L1210
;
pathology
;
Mice
;
Molecular Structure
;
Schiff Bases
;
chemical synthesis
;
chemistry
;
pharmacology
;
Triazoles
;
chemical synthesis
;
chemistry
;
pharmacology
7.Synthesis and antitumor activity of C3 heterocyclic-substituted fluoroquinolone derivatives (I): ciprofloxacin aminothiodiazole Schiff-bases.
Guo-Qiang HU ; Xiao-Kui WU ; Xin WANG ; Zhi-Qiang ZHANG ; Song-Qiang XIE ; Wen-Long HUANG ; Hui-Bin ZHANG
Acta Pharmaceutica Sinica 2008;43(11):1112-1115
To discover a novel antitumor lead compound derived from fluoroquinolone, C3 carboxyl group of ciprofloxacin (1) was replaced with heterocyclic ring to form cyclopropyl fluoroquinolone aminothiadiazole scaffold (2), then reacted with aromatic aldehydes to give the Schiff bases compounds (3a-3j). The structures of new compounds were characterized by element analysis and spectral data, and their in vitro antitumor activity against SMMC-7721, HL60 and L1210 cell lines was evaluated by MTT assay via the respective IC50 values. The bioactive assay showed that eleven thiadiazole-substituted ciprofloxacin derivatives displayed potential cytotoxicity against the tested cancer cell lines, where the IC50 values of compounds 3d and 3f reached micromolar concentration. Therefore, the C3 carboxyl group of fluoroquinolone is not necessary to antitumor activity. Functionally modified heterocycle-substituted fluoroquinolone as potent antitumor lead compound is valuable for further study.
Animals
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Antineoplastic Agents
;
chemical synthesis
;
pharmacology
;
Cell Line, Tumor
;
drug effects
;
Ciprofloxacin
;
analogs & derivatives
;
chemical synthesis
;
pharmacology
;
Drug Screening Assays, Antitumor
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HL-60 Cells
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Humans
;
Inhibitory Concentration 50
;
Leukemia L1210
;
pathology
;
Liver Neoplasms
;
pathology
;
Schiff Bases
;
chemical synthesis
;
pharmacology
8.Anti-tumor effect of two adjuvants of CpG ODN on leukemic tumor in mouse models.
Ai-Li HE ; Hong CHEN ; Wang-Gang ZHANG ; Xing-Mei CAO ; Wan-Hong ZHAO
Journal of Experimental Hematology 2007;15(5):1042-1045
To investigate the anti-tumor and side effect of CpG ODN 1826 and CpG ODN2006 as an adjuvants on leukemic tumor in mouse-models, an acute lymphocytic leukemic tumor in mouse model was established, then inoculated inactivated L1210 cells alone or with different vaccine adjuvants were injected subcutaneously into each DBA/2 model mouse at different times. The activities of mice, the tumor formation rate and the growth status of leukemic tumor were observed. The tumor was examined by pathologic section. The results showed that the vaccine of inactivated L1210 cells and CpG ODN 1826 could decrease the leukemic tumor formation rate, slow down the growth of leukemic tumor mass in mice and obviously cause necrosis of tumor cells, but it could not prolong the life spans of the tumor-burden mice; while CpG ODN2006 could not only decrease the tumor formation rate, slow down the growth of tumor mass in mice and result in obvious necrosis of tumor cells, but also could eliminate the existing tumor mass in mice, and prolong the life spans of the tumor-burden mice. It is concluded that using CpG ODN2006 as an adjuvant enhances the anti-tumor effect against the leukemic tumor, prolong the life span of tumor-burden mice without obvious side effect.
Adjuvants, Immunologic
;
therapeutic use
;
Animals
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Cancer Vaccines
;
therapeutic use
;
Cytotoxicity, Immunologic
;
immunology
;
Female
;
Leukemia L1210
;
immunology
;
therapy
;
Male
;
Mice
;
Mice, Inbred DBA
;
Neoplasm Transplantation
;
Oligodeoxyribonucleotides
;
therapeutic use
;
Random Allocation
9.Indoleamine 2, 3-dioxygenase expression in cells of human acute monocyte leukemia (M(5)) and acute lymphocyte leukemia and therapeutic effect of its inhibitor 1-methyl tryptophan.
Jing-Xin SUN ; Wang-Gang ZHANG ; Yin-Xia CHEN ; Wan-Hong ZHAO ; Wei TIAN ; Yun YANG ; Su-Hu LIU
Journal of Experimental Hematology 2007;15(3):478-482
The objective of this study was to investigate the expression and function of indoleamine 2, 3-dioxygenase (IDO) in leukemia. The IDO expressions in human acute monocyte leukemia (M(5)) and acute lymphocyte leukemia (ALL) were detected by immunofluorescence staining. Constructed leukemia mouse model was used to observe whether the IDO inhibitor, 1-methyl tryptophan (1-MT), has any effect in treating leukemia. The experimental group were fed with 1-MT solution every day while the mice in control group had no further treatment. The results showed that the average ratios of IDO expression were 29.4 +/- 11.2% in M(5) patients and 24.7 +/- 7.96% in ALL patients respectively. After statistical test, IDO expression level in leukemia cells was significantly higher than that of normal mononuclear cells. The tumor decreased gradually in mice treated with 1-MT. At the terminal point of the experiment (88 days after vaccination), the average survival time in the experimental group was 42.3 days while the mice in control group only lived 15.1 days in average, which difference was statistically significant (P < 0.05). Some of the leukemia mice in the experimental group long-term survived without tumor (more than three months after vaccination). It is concluded that human acute monocyte leukemia (M(5)) and acute lymphocyte leukemia (ALL) express IDO, and both can be treated by 1-MT in mice.
Adolescent
;
Adult
;
Animals
;
Child
;
Female
;
Humans
;
Indoleamine-Pyrrole 2,3,-Dioxygenase
;
antagonists & inhibitors
;
metabolism
;
Leukemia L1210
;
drug therapy
;
Leukemia, Biphenotypic, Acute
;
drug therapy
;
enzymology
;
Leukemia, Monocytic, Acute
;
drug therapy
;
enzymology
;
Male
;
Mice
;
Mice, Inbred DBA
;
Middle Aged
;
Tryptophan
;
analogs & derivatives
;
therapeutic use
;
Young Adult
10.Apoptosis-inducing effect of alternol on mouse lymphocyte leukemia cells and its mechanism.
Zhao-zhe LIU ; Jie-peng CHEN ; Su-lan ZHAO ; Chang-ling LI
Acta Pharmaceutica Sinica 2007;42(12):1259-1265
Alternol is purified from fermentation productions of microorganisms named as Alternaria alternata var. monosporus. The research is to investigate the apoptosis-inducing effect of alternol on mouse lymphocyte leukemia (L1210) cells and the possible mechanisms. MTT method was used to evaluate the viability of L1210 cells. Apoptosis of L1210 cells was detected by morphological assessment, DNA electrophoresis assay and flow cytometry. Western blotting analysis was carried out to determine the apoptosis-related proteins. Proliferation inhibition of L1210 cells by alternol was found remarkably in a dose-dependent manner. When treated with alternol, apoptotic morphological features of L1210 cells were observed by fluorescent microscopy (AO/EB) and the apoptosis rate was also elevated in a time-dependent manner. After treatments with various concentrations of alternol for 48 h, DNA laddering appeared. The increase of reactive oxygen species (ROS) production was found after cells were exposed to alternol for 6 h, while the decrease of mitochondrial transmembrane potential (delta psi m) was not found until cells were exposed to alternol for 24 h. Furthermore, the level of Bcel-2 and Bcl-2/Bax was down-regulated, while the level of caspase-3 and caspase-9 but not caspase-8 was up-regulated when alternol was added for 72 h. In summary, the results suggested that alternol could inhibit the proliferation of L1210 cells and induce apoptosis of L1210 cells, which was mediated by mitochondria-dependent pathway.
Alternaria
;
chemistry
;
Animals
;
Apoptosis
;
drug effects
;
Caspase 3
;
metabolism
;
Caspase 8
;
metabolism
;
Caspase 9
;
metabolism
;
Cell Line, Tumor
;
Cell Proliferation
;
drug effects
;
Dose-Response Relationship, Drug
;
Heterocyclic Compounds, 4 or More Rings
;
pharmacology
;
Leukemia L1210
;
metabolism
;
pathology
;
Membrane Potential, Mitochondrial
;
drug effects
;
Mice
;
Proto-Oncogene Proteins c-bcl-2
;
metabolism
;
Reactive Oxygen Species
;
metabolism
;
bcl-2-Associated X Protein
;
metabolism