藻蓝蛋白联合全反式维甲酸对HeLa细胞生长抑制作用的研究(英文)
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摘要
目的:探讨全反式维甲酸和藻蓝蛋白单独及联合用药对HeLa细胞生长的影响,并揭示两者联合用药对细胞周期和细胞凋亡影响的分子机制。方法:MTT法检测全反式维甲酸和藻蓝蛋白单独及联合用药对HeLa细胞生长的影响,原位杂交法检测用药前后细胞内CDK-4基因mRNA的表达情况,免疫组化法检测用药前后bcl-2基因的表达情况,TUNEL法检测用药前后细胞凋亡情况。结果:全反式维甲酸和藻蓝蛋白均具有抑制HeLa细胞生长的作用,当达到相同的抑制率时,联合藻蓝蛋白使用可以显著降低全反式维甲酸的使用剂量从而达到降低毒副作用的目的。两者联合用药可以显著降低CDK-4的表达量从而对HeLa细胞的细胞周期产生影响。两者联合用药可以显著下调bcl-2的表达水平从而引发细胞凋亡。结论:通过联合藻蓝蛋白,可以显著降低全反式维甲酸的使用剂量从而降低毒副作用。全反式维甲酸和藻蓝蛋白联合用药抑制HeLa细胞生长的分子机制可能是通过抑制CDK-4和bcl-2的表达来影响细胞周期并最终导致细胞凋亡。
Objective: We focused on studying the effects of All-transretinoic acid(ATRA), C-phycocyanin(C-PC) or ATRA +C-PC on HeLa cell growth, cell cycle distribution and apoptosis. Methods: MTT assay was adopted to determine the effects of C-PC and/or ATRA on the growth of HeLa cells. The expressions of CDK-4 and Bcl-2 were determined by in situ hybridization and immunohistochemistry staining. TUNEL assay was used to analyze the cell apoptosis. Results: Both C-PC and ATRA could inhibit the growth of HeLa cells. And these two drugs showed synergistically inhibitory effects. The dosage of ATRA was reduced when used with C-PC. ATRA, combined with C-PC, was more efficient at inducing cell cycle arrest by decreasing CDK-4 expression. The combination of the two drugs could lead to down-regulation of Bcl-2 expression and thereby cell apoptosis. Conclusion: By combined with C-PC,ATRA can be used at lower dosage with reduced toxicity. C-PC + ATRA combination might exert the inhibitory effects on cell growth by inducing cell cycle arrest and cell apoptosis.
Objective: We focused on studying the effects of All-transretinoic acid(ATRA), C-phycocyanin(C-PC) or ATRA +C-PC on HeLa cell growth, cell cycle distribution and apoptosis. Methods: MTT assay was adopted to determine the effects of C-PC and/or ATRA on the growth of HeLa cells. The expressions of CDK-4 and Bcl-2 were determined by in situ hybridization and immunohistochemistry staining. TUNEL assay was used to analyze the cell apoptosis. Results: Both C-PC and ATRA could inhibit the growth of HeLa cells. And these two drugs showed synergistically inhibitory effects. The dosage of ATRA was reduced when used with C-PC. ATRA, combined with C-PC, was more efficient at inducing cell cycle arrest by decreasing CDK-4 expression. The combination of the two drugs could lead to down-regulation of Bcl-2 expression and thereby cell apoptosis. Conclusion: By combined with C-PC,ATRA can be used at lower dosage with reduced toxicity. C-PC + ATRA combination might exert the inhibitory effects on cell growth by inducing cell cycle arrest and cell apoptosis.
引文
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[20]Nadzialek S,Pigneur LM,Weron B,et al.Bcl-2 and Caspase 3mRNA levels in the testes of gudgeon,Gobio gobio,exposed to ethinylestradiol(EE2)[J].Aquat Toxicol,2010,98(3):304-310
[2]Marquardt J,Senger H,Miyashita H,et al.Isolation and characterization of biliprotein aggregates from Acaryochloris marina,a Prochloronlike prokaryote containing mainly chlor-ophyll d[J].FEBS Lett,1997,410(2-3):428-432
[3]Thangam R,Suresh V,Asenath Princy W,et al.C-Phycocyan-in from Oscillatoria tenuis exhibited an antioxidant and in vitro antiproliferative activity through induction of apoptosis and G0/G1 cell cycle arrest[J].Food Chem,2013,140(1-2):262-272
[4]Cherng SC,Cheng SN,Tarn A,et al.Anti-inflammatory activity of c-phycocyanin in lipopolysaccharide-stimulated RAW 264.7 macrophages[J].Life Sci,2007,81(19-20):1431-1435
[5]Li B,Chu X,Gao M,et al.Apoptotic mechanism of MCF-7 breast cells in vivo and in vitro induced by photodynamic therapy with C-phycocyanin[J].Acta Biochim Biophys Sin(Shanghai),2010,42(1):80-89
[6]Bhat VB,Madyastha KM.C-phycocyanin:a potent peroxyl radical scavenger in vivo and in vitro[J].Biochem Biophys Res Commun,2000,275(1):20-25
[7]Congleton J,Jiang H,Malavasi F,et al.ATRA-induced HL-60myeloid leukemia cell differentiation depends on the CD38 cytosolic tail needed for membrane localization,but CD38 enzymatic activity is unnecessary[J].Exp Cell Res,2011,317(7):910-919
[8]Siddikuzzaman,Grace VM.Inhibition of metastatic lung cancer in C57BL/6 mice by liposome encapsulated all trans retinoic acid(ATRA)[J].Int Immunopharmacol,2012,14(4):570-579
[9]Zhao W,Wang H,Wang X,et al.Effects of all-trans-retinoic acid and arsenic trioxide on the hemostatic disturbance associated with acute promyelocytic leukemia[J].Thromb Res,2001,102(3):197-204
[10]Montillo M,Ricci F,Tedeschi A,et al.Twice daily fludarabine/AraC associated to idarubicin,G-CSF and ATRA is an effective salvage regimen in non-promyelocytic acute myeloid leukemia[J].Leuk Res,2009,33(8):1072-1078
[11]Li B,Zhang X,Gao M,et al.Effects of CD59 on antitumoral activities of phycocyanin from Spirulina platensis[J].Biomed Pharmacother,2005,59(10):551-560
[12]Tolis C,Peters GJ,Ferreira CG,et al.Cell cycle disturbances and apoptosis induced by topotecan and gemcitabine on human lung cancer cell lines[J].Eur J Cancer,1999,35(5):796-807
[13]MarzinkeMA,Clagett-DameM.Theall-transretinoicacid(atRA)-regulated gene Calmin(Clmn)regulates cell cycle exit and neurite outgrowth in murine neuroblastoma(Neuro2a)cells[J].Exp Cell Res,2012,318(1):85-93
[14]Nakagawa K,Conrad NK,Williams JP,et al.Mechanism of inactivation of CDKN2 and MTS2 in non-small cell lung cancer and association with advanced stage[J].Oncogene,1995,11(9):1843-1851
[15]Gautschi O,Ratschiller D,Gugger M,et al.Cyclin D1 in non-small cell lung cancer:a key driver of malignant transformation[J].Lung Cancer,2007,55(1):1-14
[16]Yu B,Lane ME,Pestell RG,et al.Downregulation of cyclin D1 alters cdk 4-and cdk 2-specific phosphorylation of retinoblastoma protein[J].Mol Cell Biol Res Commun,2000,3(6):352-359
[17]Sanz L,Garcia-Marco JA,Casanova B,et al.Bcl-2 family gene modulation during spontaneous apoptosis of B-chronic lymphocytic leukemia cells[J].Biochem Biophys Res Commun,2004,315(3):562-567
[18]Yan W,Suominen J,Samson M,et al.Involvement of Bcl-2 family proteins in germ cell apoptosis during testicular development in the rat and pro-survival effect of stem cell factor on germ cells in vitro[J].Mol Cell Endocrinol,2000,165(1-2):115-129
[19]Heiser D,Labi V,Erlacher M,et al.The Bcl-2 protein family and its role in the development of neoplastic disease[J].Exp Gerontol,2004,39(8):1125-1135
[20]Nadzialek S,Pigneur LM,Weron B,et al.Bcl-2 and Caspase 3mRNA levels in the testes of gudgeon,Gobio gobio,exposed to ethinylestradiol(EE2)[J].Aquat Toxicol,2010,98(3):304-310