Evaluation of tumor formation of three bladder cancer cell lines in nude mice

Abstract

This study examined the differences in tumor formation of three bladder tumor cell lines (BIU-87, T24 and EJ) after subcutaneously transplanted into nude mice, in order to find the best technique for establishing in vivo bladder tumor model. BIU-87, T24 and EJ cells at logarithmic phase were re-suspended in serum-free medium. The cells suspensions of the identical concentration were subcutaneously transplanted into nude mice and then the success rate and tumor growth were compared among the three cell groups. The results of tumor formation were pathologically evaluated. Lung, liver and kidney tissues were also pathologically examined for distant metastasis. The proliferation of the three cells were determined by immunohistochemically detecting the PCNA expression in the tumors. The results showed that the success rates of EJ and T24 cells were significantly higher than that of BIU-87 cells and no distant metastasis was noted among the three groups. The proliferation levels of EJ and T24 cells was significantly higher than that of BIU-87. But at the later stage of tumor formation, as compared with T24 cells, EJ grew more vigorously, soon resulting in the central necrosis of tumor, which affected the measurement of the actual size of the tumors. Moreover, PCNA staining exhibited that the proliferation of EJ and T24 was significantly higher than that of BIU-87 cells. It is concluded that as compared with BIU-87 cells, EJ and T24 cells had higher success rates, with not significant differences in death rate and distant metastasis found among them. There existed no significant difference in tumor formation between EJ and T24 cells and T24 cells do not rupture easily, which makes it a better cell line for the establishment of in vivo bladder tumor model.

Keywords

bladder cancer / nude mice / subcutaneous tumor formation

References

[1] HuH.B., LuoC.L.. Advances in studies of animal models for bladder tumors. Int J Lab Med (Chinese), 2004, 25(5): 426-428

[2] KellandL.R.. Of mice and men: values and liabilities of the athymic nude mouse model in anticancer drug development. Eur J Cancer, 2004, 40(6): 827-836

[3] KerbelR.S.. Human tumor xenografts as predictive preclinical models for anticancer drug activity in humans: better than commonly perceived-but they can be improved. Cancer Biol Ther, 2003, 2(4Suppl1): S134-S139

[4] CéspedesM.V., CasanovaI., ParreñoM., et al.. Mouse models in oncogenesis and cancer therapy. Clin Transl Oncol, 2006, 8(5): 318-329

[5] SinghM., LimaA., MolinaR., et al.. Assessing therapeutic responses in Kras mutant cancers using genetically engineered mouse models. Nat Biotechnol, 2010, 28(6): 585-593

[6] MelquistJ.J., KackaM., LiY., et al.. Conditionally replicating adenovirus-mediated gene therapy in bladder cancer: an orthotopic in vivo model. Urol Oncol, 2006, 24(4): 362-371

[7] ChongL., RupingY., JianchengB., et al.. Characterization of a novel transplantable orthotopic murine xenograft model of a human bladder transitional cell tumor (BIU-87). Cancer Biol Ther, 2006, 5(4): 394-398

[8] LiangP.H., JinF.S., ZhangK.Q., et al.. Establishment of nude mouse transplantation tumor model with human bladder cancer and the study of tumor angiogenesis. J Clin Urol (Chinese), 2006, 21(9): 711-13

[9] ZhangL.L., HeD.L., LiX., et al.. Overexpression of coxsackie and adenovirus receptor inhibit growth of human bladder cancer cell in vitro and in vivo. Acta Pharmacol Sin, 2007, 28(6): 895-900

[10] ZiX.L., SimoneauA.R., FlavokawainA.. a novel chalcone from kava extract, induces apoptosis in bladder cancer cells by involvement of Bax protein-dependent and mitochondria dependent apoptotic pathway and suppresses tumor growth in mice. Cancer Res, 2005, 65(8): 3479-3486

[11] BlackP.C., DinneyC.P.. Bladder cancer angiogenesis and metastasis translation from murine model to clinical trial. Cancer Metastasis Rev, 2007, 26(3–4): 623-634

[12] MitraA.P., DatarR.H., CoteR.J.. Molecular pathways in in vasive bladder cancer: new insights into mechanisms, progression, and target identification. J Clin Oncol, 2006, 24(35): 5552-5564

[13] ZhangB.L., XuM.L., GaoM.T., et al.. Methodological study on improving tumor formation rate of xenografted human lung cancer in nude mice. Chin J Clin Pharmacol Ther (Chinese), 2007, 12(9): 1052-1057

[14] XuX., PrestwichG.D.. Inhibition of tumor growth and angiogenesis by a lysophosphatidic acid antagonist in an engineered three-dimensional lung cancer xenograft model. Cancer, 2010, 116(7): 1739-1750

[15] KhannaC., HunterK.. Modeling metastasis in vivo. Carcinogenesis, 2005, 26(3): 513-523

[16] HernandezL., SmirnovaT., WyckoffJ., et al.. In vivo assay for tumor cell invasion. Methods Mol Biol, 2009, 571: 227-238