Somers Kenneth D, Brown Roy R, Holterman Daniel A, Yousefieh Nazita, Glass William F, Wright George L, Schellhammer Paul F, Qian Jesse, Ciavarra Richard P
Departments of Microbiology and Molecular Cell Biology, Eastern Virginia Medical School, Norfolk, VA 23507, USA.
Int J Cancer. 2003 Dec 10;107(5):773-80. doi: 10.1002/ijc.11464.
We established an orthotopic treatment model of prostate cancer to generate reproducible primary and metastatic carcinoma in immunocompetent C57BL/6 mice. Using an in vivo selection scheme of intraprostatic implantation of TRAMP-C1 cells, primary prostate tumors were cultured and recycled three times by intraprostatic injection resulting in the selection and establishment of the recycled cell line TRAMP-C1P3. Prostate tumors were detected approximately 30 days post-implantation with periaortic lymph node metastasis in 19/20 (95%) of mice. Tissue culture amplification, DNA ploidy and PCR amplification of the SV40 transgene were used to detect metastatic TRAMP-C1P3 in lymph node specimens. Tissue culture amplification and DNA ploidy were as sensitive as SV40 transgene amplification by PCR in detection of early metastatic disease in draining lymph nodes. To establish the use of the orthotopic model of prostate cancer for immunotherapy, mice were injected orthotopically with TRAMP-C1P3 cells and 7 days post-implantation treated daily for 28 days with either flt3L or carrier control. Carrier-treated mice had clinically detectable prostate tumors, lymph node metastasis and were moribund at 29-35 days, whereas flt3L therapy markedly suppressed primary TRAMP-C1P3 growth and lymph node metastasis, and prolonged survival. In summary, we have established a reproducible and clinically relevant orthotopic treatment model of prostate cancer in immunocompetent mice with application to a variety of therapeutic strategies. We demonstrate that flt3L treatment suppressed orthotopic prostate tumor growth and lymph node metastasis reinforcing a role for flt3L as an immunotherapeutic strategy for prostate cancer.
我们建立了一种前列腺癌原位治疗模型，以在具有免疫活性的C57BL/6小鼠中产生可重复的原发性和转移性癌。通过前列腺内植入TRAMP-C1细胞的体内选择方案，培养原发性前列腺肿瘤，并通过前列腺内注射进行三次循环，从而选择并建立了循环细胞系TRAMP-C1P3。植入后约30天检测到前列腺肿瘤，20只小鼠中有19只（95%）出现主动脉旁淋巴结转移。利用组织培养扩增、DNA倍性分析以及SV40转基因的PCR扩增来检测淋巴结标本中的转移性TRAMP-C1P3。在检测引流淋巴结中的早期转移性疾病时，组织培养扩增和DNA倍性分析与通过PCR进行的SV40转基因扩增一样敏感。为了建立前列腺癌原位模型用于免疫治疗，将TRAMP-C1P3细胞原位注射到小鼠体内，植入后7天，每天用flt3L或载体对照处理28天。接受载体处理的小鼠出现临床上可检测到的前列腺肿瘤、淋巴结转移，并在29 - 35天濒死，而flt3L治疗显著抑制了原发性TRAMP-C1P3的生长和淋巴结转移，并延长了生存期。总之，我们在具有免疫活性的小鼠中建立了一种可重复且与临床相关的前列腺癌原位治疗模型，可应用于多种治疗策略。我们证明flt3L治疗抑制了原位前列腺肿瘤的生长和淋巴结转移，强化了flt3L作为前列腺癌免疫治疗策略的作用。
