靶向端粒G-四链体手性钌配合物的抗肿瘤活性研究
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摘要
[目的]研究手性钌配合物Λ-[Ru(bpy)2(pyip)]2+(Λ-OMe)、Δ-[Ru(bpy)2(pyip)]2+(Δ-OMe)及其外消旋化合物[Ru(phen)2p-MOPIP]2+(dl-OMe)在体外实验中的抗肿瘤活性。[方法]3种配合物分别作用于人胃癌细胞株(MGC-803),人结肠癌细胞株(Colo205),人乳腺癌细胞株(MCF-7),人肺腺癌上皮细胞株(A549),人肝癌细胞株(Hep G2),人舌鳞状细胞癌株(SCC-9)48 h,以人胃黏膜上皮细胞株(GES-1)为正常对照细胞株,顺铂为阳性对照药物,四甲基偶氮唑蓝(MTT)法筛选出其中半抑制浓度(IC50)最小的药物和药敏作用最好的肿瘤细胞。筛选出药物和肿瘤细胞后,再用MTT法测定24、48、72 h相应药物对肿瘤细胞的抑制作用;Hoechest33342染色法测定其对肿瘤细胞的凋亡诱导作用。[结果]MTT法筛选出Λ-OMe对胃癌MGC-803细胞的增殖抑制作用最好,即(IC50)最小,在作用48 h的半数抑制浓度为:7.4 mg/L,且呈现很好的时间和剂量依赖性。Hoechest33342染色显示,在药物处理48 h后,随着药物浓度增加,药物促凋亡作用越明显。[结论]手性钌配合物具有良好的抗肿瘤活性,其中以Λ-OMe抗肿瘤活性最好。
[Objective] Study on anti-tumor activities research of a paire of chiral ruthenium complexes Λ-[Ru(bpy)2(pyip)]2 +(Λ-OMe)、Δ-[Ru(bpy)2(pyip)]2 +(Δ-OMe)and their racemic mixture [Ru(phen)2p-MOPIP]2 +(dl-OMe)in vitro experiments. [Methods] Three complexes were acting 48 hours on human gastric cancer cell line(MGC-803), human colon cancer cell lines(Colo205), human breast cancer cell line(MCF-7), human lung adenocarcinoma epithelial cell line( A549), human hepatocellular carcinoma cell line( Hep G2), human tongue squamous cell carcinoma lines(SCC-9), human gastric epithelial cell line(GES-1) and cisplatin(CDDP) is a positive control.And then find the drug and tumor cells which half inhibitory concentration(IC50) was the smallest by MTT assay. Second,Detect the drug's inhibitory effect on tumor cells after drug treatment 24 、48、72 hours. Third, induced apoptosis of gastric cancer cells with Hoechest33342 staining. [Results] The best anti-tumor effects drug and cell is chiral ruthenium complexes Λ- [Ru(phen) 2p-MOPIP]2+(Λ-OMe) and MGC-803 with MTT assay. The IC50is:7.4 μg/m L at48 hours.The tested compound induces cell death in a dose and time dependent manner on MGC-803 cells;Hoechest33342 staining showed that with the increase of drug concentration,more obvious of apoptosis after 48 hours drug treatment. [Conclusion] Chiral ruthenium complexes have good anti-tumor activity and the Λ-OMe have significant anti-tumor activities.
[Objective] Study on anti-tumor activities research of a paire of chiral ruthenium complexes Λ-[Ru(bpy)2(pyip)]2 +(Λ-OMe)、Δ-[Ru(bpy)2(pyip)]2 +(Δ-OMe)and their racemic mixture [Ru(phen)2p-MOPIP]2 +(dl-OMe)in vitro experiments. [Methods] Three complexes were acting 48 hours on human gastric cancer cell line(MGC-803), human colon cancer cell lines(Colo205), human breast cancer cell line(MCF-7), human lung adenocarcinoma epithelial cell line( A549), human hepatocellular carcinoma cell line( Hep G2), human tongue squamous cell carcinoma lines(SCC-9), human gastric epithelial cell line(GES-1) and cisplatin(CDDP) is a positive control.And then find the drug and tumor cells which half inhibitory concentration(IC50) was the smallest by MTT assay. Second,Detect the drug's inhibitory effect on tumor cells after drug treatment 24 、48、72 hours. Third, induced apoptosis of gastric cancer cells with Hoechest33342 staining. [Results] The best anti-tumor effects drug and cell is chiral ruthenium complexes Λ- [Ru(phen) 2p-MOPIP]2+(Λ-OMe) and MGC-803 with MTT assay. The IC50is:7.4 μg/m L at48 hours.The tested compound induces cell death in a dose and time dependent manner on MGC-803 cells;Hoechest33342 staining showed that with the increase of drug concentration,more obvious of apoptosis after 48 hours drug treatment. [Conclusion] Chiral ruthenium complexes have good anti-tumor activity and the Λ-OMe have significant anti-tumor activities.
引文
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[18]Nitha LP,Aswathy R,Mathews NE,et al.Synthesis,spectroscopic characterisation,DNA cleavage,superoxidase dismutase activity and antibacterial properties of some transition metal complexes of a novel bidentate Schiff base derived from isatin and 2-aminopyrimidine[J].Spectrochim Acta A Mol Biomol Spectrosc,2014,118:154-161.
[19]Sun D,Zhang W,Lv M,et al.Antibacterial activity of ruthenium(II)polypyridyl complex manipulated by membrane permeability and cell morphology[J].Bioorg Med Chem Lett,2015,25(10):2068-2073.
[20]Malipeddi M,Lakhani C,Chhabra M,et al.An efficient synthesis and in vitro antibacterial evaluation of rutheniumquinolinol complexes[J].Bioorg Med Chem,2015,25(15):2892-2896.
[2]Mendoza-Ferri MG,Hartinger CG,Mendoza MA,et al.Transferring the concept of multinuclearity to rutheum complexes for im Provement of anticancer activity[J].Med Chem,2009,52(4):916-925.
[3]Heffeter P,Jungwirth U,Jaku Pec M,et al.Resistance agains tnovel anticancer metal compounds:differences and similarities[J].Drug Resist UPdat,2008,11(l-2):1-16.
[4]张小年,刘亚楠,杨晓新,等.钌配合物诱导肿瘤细胞凋亡的信号通路及其作用机制[J].化学进展,2011,21(5):983-990.
[5]Stevens SK,Strehle AP,Miller RL,et al.The anticancer ruthenium complex KP1019 induces DNA damage,leading to cell cycle delay and cell death in Saccharomyces cerevisiae[J].Molecular Pharmacology,2013,83(1):225-234.
[6]Chakree K,Ovatlarnporn C,Dyson PJ,et al.Altered DNA Binding and Amplification of Human Breast Cancer Suppressor Gene BRCA1 Induced by a Novel Antitumor Compound,[Ru(η6-p-phenylethacrynate)Cl2(pta)][J].International Journal of Molecular Sciences,2012,13(10):13183-13202.
[7]Heffeter P,Atil B,Kryeziu K,et al.The ruthenium compound KP1339 potentiates the anticancer activity of sorafenib in vitro and in vivo[J].European Journal of Cancer,2013,49(15):3366-3375.
[8]Rehmann JP,Barton JK.Proton NMR studies of tris(phenanthroline)metal complexes bound to oligonucleotides:characterization of binding modes[J].Biochemistry,1990,29,1701-1709.
[9]Eriksson MM,Leijon M,Hiort C,et al.Minor groove binding of[Ru(phen)3]2+to[d(CGCGATCGCG)]2 evidenced by two-dimensional NMR[J].J.Am.Chem.Soc,1992,114:493-494.
[10]Mei WJ,Liu J,Zheng KC,et al.Experimental and theoretical study on DNA-binding andphotocleavage properties of chiral complexesΔ-andΛ-[Ru(bpy)2 L](L=o-hpip,m-hpip and p-hpip)[J].Dalton Trans,2003,32(7):1352-1359.
[11]Kumar CV,Barton JK,Turro N J.Photophysics ofruthenium complexes bound to double helical DNA[J].J Am Chem Soc,1985,107:5518-5520.
[12]刘亚楠,杨芳,梅文杰,等.手性钌配合物的合成、抗肿瘤活性及其与血清蛋白的相互作用[J].高等学校化学学报,2010,31(3):435-441.
[13]Shi S,Geng XT,Zhao J,et al.Interaction of[Ru(bpy)2(dppz)]2+with human telomeric DNA:Preferential binding to G-quadruplexes over i-motif[J].Biochimie,2010,92:370-377.
[14]Sun D,Liu Y,Liu D,et al.Stabilization of G-quadruplex DNA,inhibition of telomerase activity and live cell imaging studies of chiral ruthenium(II)complexes[J].Chemistry,2012,18(14):4285-4295.
[15]Mosmann T.Rapid colormetric assay for cellular growth and survial:applocation to proliferation and cytotoxicity assays[J].Immand Methods,1983,65:55-58.
[16]宋恩峰,张珏.消化道肿瘤的中医药治疗进展[C].中国中西医结合学会肿瘤专业委员会第三届国际中医、中西医结合肿瘤学术交流大会暨第十二届全国中西医结合肿瘤学术大会,2010:4.
[17]Pahontu E,Julea F,Rosu T,et al.Antibacterial,antifungal and in vitro antileukaemia activity of metal complexes with thiosemicarbazones[J].Cell Mol Med,2015,19(4);865-878.
[18]Nitha LP,Aswathy R,Mathews NE,et al.Synthesis,spectroscopic characterisation,DNA cleavage,superoxidase dismutase activity and antibacterial properties of some transition metal complexes of a novel bidentate Schiff base derived from isatin and 2-aminopyrimidine[J].Spectrochim Acta A Mol Biomol Spectrosc,2014,118:154-161.
[19]Sun D,Zhang W,Lv M,et al.Antibacterial activity of ruthenium(II)polypyridyl complex manipulated by membrane permeability and cell morphology[J].Bioorg Med Chem Lett,2015,25(10):2068-2073.
[20]Malipeddi M,Lakhani C,Chhabra M,et al.An efficient synthesis and in vitro antibacterial evaluation of rutheniumquinolinol complexes[J].Bioorg Med Chem,2015,25(15):2892-2896.