Inhibitory effects of TNP-470 in combination with BCNU on tumor growth of human glioblastoma xenografts

Abstract

This study investigated the effect of TNP-470 in combination with carmustine (BCNU) on the growth of subcutaneously implanted human glioblastoma xenografts in nude mice. Human glioblastoma U-251 cells (1×10⁷) were injected into 24 nude mice subcutaneously. The tumor-bearing mice were randomly divided into 4 groups on the seventh day following tumor implantation: TNP-470 group, in which TNP-470 was given 30 mg/kg subcutaneously every other day 7 times; BCNU group, in which 20 mg/kg BCNU were injected into peritoneal cavity per 4 days 3 times; TNP-470 plus BCNU group, in which TNP-470 and BCNU were coadministered in the same manner as in the TNP-470 group and the BCNU group; control group, in which the mice were given 0.2 mL of the mixture including 3% ethanol, 5% acacia and 0.9% saline subcutaneously every other day 7 times. The tumor size and weights were measured. The tumor microvessel density (MVD) was determined by immunostaining by using goat-anti-mouse polyclonal antibody CD105. The results showed that on the 21th day following treatment, the volume of xenografts in the TNP-470 plus BCNU group was (108.93±17.63)mm³, markedly lower than that in the TNP-470 group [(576.10±114.29)mm³] and the BCNU group [(473.01±48.04)mm³] (both P<0.01). And the xenograft volume in these 3 treatment groups was even much lower than that in the control group [(1512.61±470.25) mm³] (all P<0.01). There was no significant difference in the volume of xenografts between the TNP-470 group and the BCNU group (P>0.05). The inhibition rate of the tumor growth in the TNP-470 plus BCNU group was (92.80±11.37)%, notably higher than that in the TNP-470 group [(61.91±6.29)%] and the BCNU group [(68.73±9.65)%] (both P<0.01) on the 21th day following treatment. There was no significant difference in the inhibition rate of tumor growth between the TNP-470 group and the BCNU group (P>0.05). The MVD of xenografts in the TNP-470 plus BCNU group was decreased significantly as compared with that in the TNP-470 group or the BCNU group (both P<0.05). The MVD of xenografts in the 3 treatment groups was markedly reduced as compared with that in the control group (all P<0.05). No significant changes in weights were observed before and after the treatment in each group (all P>0.05). It was concluded that the combination of TNP-470 and BCNU can significantly inhibit the growth of human glioblastoma xenografts in nude mice without evident side effects.

Keywords

angiogenesis / antiangiogenesis / TNP-470 / endoglin(CD105) / microvessel density

References

[1] FolkerthR.D.. Descriptive analysis and quantification of angiogenesis in human tumors. J Neurooncol, 2000, 50(1–2): 165-172

[2] PuduvalliU.K., SawagaR.. Antiangiogenesis therapeutic strategies and clinical implications for brain tumors. J Neurooncol, 2000, 50(1–2): 189-200

[3] KraghM., Spang-ThomsenM., KristjansenP.E.. Time until initiation of tumor growth is an effective measure of the anti-angiogenic effect of TNP-470 on human glioblastoma in nude mice. Oncol Rep, 1999, 6(4): 759-762

[4] LinlinD., XiuwuB., YishengC., et al.. Inhibitory effects of angiogenesis inhibitor TNP-470 on growth of a human malignant glioma cell line SHG-44 and its xenografted tumor. Acta Academiae Medicine Militaris Tertiae, 2001, 23(3): 272-275

[5] TakamiyaY., BremH., OjeifoJ., et al.. AGM-1470 inhibits the growth of human glioblastoma cells in vitro and in vivo. Neurosurg, 1994, 34(5): 869-875

[6] BernsenH.J., RijkenP.F., PetersH., et al.. The effect of the anti-angiogenic agent TNP-470 on tumor growth and vascularity in low passaged xenografts of human gliomas in nude mice. J Neurooncol, 1998, 38(1): 51-57

[7] LundE.L., BastholmL., KristjansenP.E.. Therapeutic synergy of TNP-470 and ionizing radiation: effects on tumor growth, vessel morphology, and angiogenesis in human glioblastoma multiforme xenografts. Clin Cancer Res, 2000, 6(3): 971-978

[8] JinglinX., BinghuiY., ShenglongY., et al.. An experimental study of the inhibitory effect of the combination of TNP-470 and MMC on the growth and metastasis of human hepatocellular carcinoma. Chin J Cancer (Chinese), 2000, 19(4): 341-343

[9] WeidnerN.. Current pathologic methods for measuring intratumoral microvessel density with breast carcinoma and other solid tumors. Breast Cancer Res Treat, 1995, 36: 169-175

[10] BennyO., NakaiK., YoshimuraT., et al.. Broad spectrum antiangiogenic treatment for ocular neovascular diseases. PLoS One, 2010, 5(9): 12515

[11] BäckmanU., SvenssonA., ChristoffersonR.H., et al.. The bisphosphonate, zoledronic acid reduces experimental neuroblastoma growth by interfering with tumor angiogenesis. Anticancer Res, 2008, 28(3A): 1551-1557

[12] BennyO., FainaruO., AdiniA., CassiolaF., et al.. An orally delivered small-molecule formulation with antiangiogenic and anticancer activity. Nat Biotechnol, 2008, 26(7): 799-807

[13] AntoineN., GreimersR., De RoanneC., et al.. AGM-1470, a potent angiogenesis inhibitor, prevents the entry of normal but not transformed endothelial cells into G1 phase of the cell cycle. Cancer Res, 1994, 54(8): 2073-2076

[14] SinN., MengL., WangM., et al.. The antiangiogenic agent fumagillin covalently binds and inhibits the methionine aminopeptidase, MetAP-2. Proc Natl Acad Sci USA, 1997, 94(12): 6099-6103

[15] KayaM., WadaT., NagoyaS., et al.. TNP-470 suppresses the tumorigenicity of HT1080 fibrosarcoma tumor through the inhibition of VEGF secretion from the tumor cells. Sarcoma, 2001, 5(4): 197-202

[16] CheslerL., GoldenbergD.D., SealesI.T., et al.. Malignant progression and blockade of angiogenesis in a murine transgenic model of neuroblastoma. Cancer Res, 2007, 67(19): 9435-9442

[17] YanaseT., TamuraM., FujitaK., et al.. Inhibition effect of angiogenesis inhibitor TNP-470 on tumor growth and metastasis of human cell lines in vitro and in vivo. Cancer Res, 1993, 53(11): 2566

[18] KudelkaA.P., VerschraegenC.F., LoverE.. Complete remission of metastatic cervical cancer with the angiogenesis inhibitor TNP-470. N Engl J Med, 1998, 338(14): 991-992

[19] DezubeB.J., Von RoennJ.H., Holden-WiltseJ.. Fumagillin analog in the treatment of Kaposi’s sarcoma: a phase I AIDS Clinical Trial Group study. AIDS Clinical Trial Group No. 215 Team. J Clin Oncol, 1998, 16(4): 1444-1449

[20] HerbstR.S., MaddenT.L., TranH.T., et al.. Safety and pharmacokinetic effects of TNP-470, an angiogenesis inhibitor, combined with paclitaxel in patients with solid tumors: evidence for activity in non-small-cell lung cancer. J Clin Oncol, 2002, 20(22): 4440-4447

[21] KudelkaA.P., LevyT., VerschraegenC.F., et al.. A phase I study of TNP-470 administered to patients with advanced squamous cell cancer of the cervix. Clin Cancer Res, 1997, 3(9): 1501-1505

[22] PradosM.D., RussoC.. Chemotherapy for brain tumors. Semin Surg Oncol, 1998, 14(1): 88-95

[23] NishimuraK., OhnoM., Chung-KangC., et al.. Anti-tumor effect of angiogenesis inhibitor TNP-470 and 5-FU combined therapy on human gastric cancer xenograft. Hepatogastroenterology, 2008, 55(82–83): 774-778

[24] WangX.F., TuL.F., WangL.H., et al.. Inhibition of expression of vascular endothelial growth factor and its receptors in pulmonary adenocarcinoma cell by TNP-470 in combination with gemcitabine. J Zhejiang Univ Sci B, 2006, 7(10): 837-843

[25] MuramakiM., MiyakeH., HaraI., et al.. Synergistic inhibition of tumor growth and metastasis by combined treatment with TNP-470 and docetaxel in a human prostate cancer PC-3 model. Int J Oncol, 2005, 26(3): 623-628

[26] MuramakiM., MiyakeH., HaraI., et al.. Synergistic inhibition of tumor growth and metastasis by combined treatment with TNP-470 and gemcitabine in a human bladder cancer KoTCC-1 model. J Urol, 2004, 172(4Pt1): 1485-1489

[27] ShweikiD., ItinA., SofferD., et al.. Vascular endothelial growth factor induced by hypoxia may mediate hypoxia-initiated angiogenesis. Nature, 1992, 359(6398): 843-845