, ON YX2015. Journal of Zhejiang Sci2Tech U niversity Vol. 27, No. 4, J ul % 60 %, 015,, : (2010)

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1 , 27, 4, Journal of Zhejiang Sci2Tech U niversity Vol. 27, No. 4, J ul : (2010) ONYX2015,,,,,, (, ) : (dichloroacetate,dca) ON YX2015, M TT DCA ON YX2015 HCT116 SW620 H T29 L02 ; Hoechst ; :DCA ON YX2015 4d HCT116 SW %, H T29 54 %, L02 DCA ON YX2015 : ON YX2015 ; ; : Q789 : A 0,, [1 ],, 5 40 % 60 %, [2 ], ON YX ON YX2015, E1B 552kD, [3 ] ON YX2015 [425 ],,, ON YX2015 Bonnet [6 ] (DCA),,,, DCA, ON YX2 015,, : : ( ) ; (SW YX0815) : ( ),,,, :, edu. cn

2 4 : ON YX ON YX2015 ; (DCA) Sigma2Aldrich ; HCT116 SW620 H T29 L02 ; (M T T) Sigma ; Ameresco ; 5 % CO2,37, 10 %FBS DM EM (Dulheccoπs Modified Eagleπs Medium) M T T ( ), HCT116 SW620 H T29 L / 96, 100 L 37 5 % CO2 24 h 10 mm DCA 10 MOI ON YX mm DCA + 10 MOI ON YX2015, 5 ( ), ( ) 37 5 % CO2 4 d, M T T (5 mg/ ml) 20 L, 37 4 h, DMSO 150 L/, 10 min, 595 nm (A595 ) (1) : OD - OD = OD - OD 100 % (1) Hoechst HCT / 96, 100 L 37 5 % CO2 24 h 10 mm DCA,10 MOI ON YX mm DCA + 10 MOI ON YX2015, 3 d, Hoechst ( 1 g/ ml), 37 5 % CO2 20 min, HCT116 L / 12, 1 ml 24 h ( ), 37 5 % CO2 5 d, 500 L (2 % 20 % ), 15 min, Excel 2003, 3, M T T DCA ON YX2015 1, 4 d, HCT116 SW %, DCA ON YX % 50 %, H T29 50 %, 4 d 30 %,ON YX2015 DCA, 20 %, L02, 2. 2 Hoechst Hoechst 33342,DCA ON YX2015 HCT116, DCA ON YX2015, ( 2)

3 , 5 d, 3,DCA ON YX2 015 HCT116, ON YX2015 DCA, ON YX2015 (10 MO I) 5 d,,, ON YX2015 HCT116, ON YX2015 DCA,, L02 10 mm DCA 10 MOI ON YX mm DCA + 10 MOI ON2 YX2015 HCT116 L02,5 d 15 min 3

4 4 : ON YX ,,,,, [7 ],,,,, ON YX2015, dl1520,, 1987 Barker Berk, p53, ON YX2015, [ 829 ], ON YX2015, p53 [ 10 ],OπShea [11 ] ON YX2015 E1B55 K RNA, p53, ON YX2015 RNA, ON YX2015,,,, :,,,,,,,, 90,,, ,S. Bonnet DCA,, ( PD K),, 2CoA,,,, 1970,DCA 2 [12 ], [13 ], FDA DCA,,,, DCA ON YX2015 HCT116 SW620 H T29, M T T, HCT116 SW620 4d 25 %, L02 DCA ON YX2015 ON YX2015 DCA, ON YX2015 DCA,,,DCA ON YX2015 ON YX2015,,, : [1 ] Jemal A, Siegel R, Ward E, et al. Cancer statistics, 2008[J ]. CA Cancer J Clin, 2008, 56 : [2 ] Mocellin S, Lise M, Nitti D. Targeted therapy for colorectal cancer : mapping the way[j ]. Trends in Molecular Medicine, 2005, 11 (7) :

5 [ 3 ] Kirn D. Clinical research result s with dl1520 ( Onyx2015), a replication2selective adenovirus for the treatment of cancer : what have we learned? [J ]. Gene Therapy, 2001, 8 (2) : [ 4 ] Khuri F R, Nemunaitis J, Ganly I, et al. A controlled trial of intratumoral ON YX2015, a selectively2replicating adenovi2 rus, in combination with cisplatin and 52fluorouracil in patient s with recurrent head and neck cancer[j ]. Nature Medicine, 2000, 6 (8) : [5 ] Heise C, Sampson2Johannes A, Williams A, et al. ON YX2015, an E1B gene2attenuated adenovirus, causes tumor2specific cytolysis and antitumoral efficacy that can be augmented by standard chemotherapeutic agent s[j ]. Nature Medicine, 1997, 3 (6) : [6 ] Bonnet S, Archer S L, Allalunis2Turner J, et al. A mitochondria2k + channel axis is suppressed in cancer and its normali2 zation promotes apoptosis and inhibits cancer growth[j ]. Cancer Cell, 2007, 11 (1) : [7 ] Alemany R, Balague C, Curiel D T. Replicative adenoviruses for cancer therapy[j ]. Nature Biotechnology, 2000, 18 (7) : [8 ] Ganly I, Kirn D, Eckhardt G, et al. A phase I study of Onyx2015, an E1B attenuated adenovirus, administered intratu2 morally to patients with recurrent head and neck cancer[j ]. Clin Cancer Res, 2000, 6 (3) : [9 ] Kirn D, Hermiston T, McCormick F. ON YX2015 : clinical data are encouraging[j ]. Nature Medicine, 1998, 4 (12) : [ 10 ] Rothmann T, Hengstermann A, Whitaker N J, et al. Replication of ON YX2015, a potential anticancer adenovirus, is in2 dependent of p53 status in tumor cells[j ]. Journal of Virology, 1998, 72 (12) : [11 ] OπShea C C, Johnson L, Bagus B, et al. Late viral RNA export, rather than p53 inactivation, determines ON YX2015 tumor selectivity[j ]. Cancer Cell, 2004, 6 (6) : [ 12 ] Stacpoole P W, Felt s J M. Diisopropylammonium dichloroacetate (DIPA) and sodium dichloracetate (DCA) : effect on glu2 cose and fat metabolism in normal and diabetic tissue[j ]. Metabolism : Clinical and Experimental, 1970, 19 (1) : [ 13 ] Stacpoole P W, Henderson G N, Yan Z, et al. Clinical pharmacology and toxicology of dichloroacetate[j ]. Environmen2 tal Health Perspectives, 1998, 106 (4) : Enhancement of Killing Effect on Colorectal Tumor Cells by Combining ONYX2015 with Dichloroacetate (DCA) X IA O L ian2li, X I E Guo2liang, L OU J un, L I X in, N IU N a, Q IA N J ing, W A N G Yi2gang ( Xinyuan Institute of Medicine and Biotechnology, Zhejiang Sci2Tech University, Hangzhou , China) Abstract : The experiment is aimed at st udying t he anti2t umor effect in colorectal t umor cells by combi2 ning ON YX2015 wit h a small molecule dichlo roacetate ( DCA ). Fir st, M T T assay is used to test t he growt h inhibition effect s of single or combination t herapy on t umor cell lines HCT116, SW620, H T29 and human normal cell line L02. Second, Hoechst is added after administration in t umor cells HCT116, and 48 hours later cells are observed under fluorescent microscope. Cytopat hic effect assay t hen is conduc2 ted to f urther demonstrate t he efficacy of t he combination t herapy. Result s showed that t he anti2tumor ef2 ficacy of ON YX2015 is significantly enhanced in combination wit h dichloroacetate, while showes no over2 lapping toxicity against normal cell line L02. Crystal violet staining also demonstrates t hat t he combination t herapy showes better efficacy t han single therapy wit h DCA or ON YX2015, respectively. Key words : ON YX2015 ; dichloroacetate ; colorectal cancer ( : )