Dong Z, Greene G, Pettaway C, Dinney C P, Eue I, Lu W, Bucana C D, Balbay M D, Bielenberg D, Fidler I J
Department of Cancer Biology, The University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Cancer Res. 1999 Feb 15;59(4):872-9.
We determined whether the IFN-beta gene can be used to suppress angiogenesis, tumor growth, and metastasis of human prostate cancer cells growing in the prostate of nude mice. Highly metastatic PC-3M human prostate cancer cells were engineered to constitutively produce murine IFN-beta subsequent to infection with a retroviral vector containing murine IFN-beta cDNA. Parental (PC-3M-P), control vector-transduced (PC-3M-Neo), and IFN-beta-transduced (PC-3M-IFN-beta) cells were injected into the prostate (orthotopic) or subcutis (ectopic) of nude mice. PC-3M-P and PC-3M-Neo cells produced rapidly growing tumors and regional lymph node metastases, whereas PC-3M-IFN-beta cells did not. PC-3M-IFN-beta cells also suppressed the tumorigenicity of bystander nontransduced prostate cancer cells. PC-3M-IFN-beta cells produced small tumors (3-5 mm in diameter) in nude mice treated with anti-asialo GM1 antibodies and in severe combined immunodeficient/Beige mice. Immunohistochemical staining revealed that PC-3M-IFN-beta tumors were homogeneously infiltrated by macrophages, whereas control tumors contained fewer macrophages at their periphery. Most tumor cells in the control tumors were stained positive by an antibody to proliferative cell nuclear antigen; very few were positively stained by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling. In sharp contrast, PC-3M-IFN-beta tumors contained fewer proliferative cell nuclear antigen-positive cells and many terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling-positive cells. Staining with antibody against CD31 showed that control tumors contained more blood vessels than PC-3M-IFN-beta tumors. PC-3M-IFN-beta cells were more sensitive to lysis mediated by natural killer cells in vitro or to cytostasis mediated by macrophages than control transduced cells. Conditioned medium from PC-3M-IFN-beta cells augmented splenic cell-mediated cytolysis to control tumor cells, which could be neutralized by antibody against IFN-beta. Collectively, the data suggest that the suppression of tumorigenicity and metastasis of PC-3M-IFN-beta cells is due to inhibition of angiogenesis and activation of host effector cells.
我们确定了干扰素-β（IFN-β）基因是否可用于抑制裸鼠前列腺中生长的人前列腺癌细胞的血管生成、肿瘤生长和转移。在感染含有鼠IFN-β cDNA的逆转录病毒载体后，对高转移性PC-3M人前列腺癌细胞进行基因改造，使其组成性产生鼠IFN-β。将亲代细胞（PC-3M-P）、对照载体转导的细胞（PC-3M-Neo）和IFN-β转导的细胞（PC-3M-IFN-β）注射到裸鼠的前列腺（原位）或皮下（异位）。PC-3M-P和PC-3M-Neo细胞产生快速生长的肿瘤和区域淋巴结转移，而PC-3M-IFN-β细胞则不会。PC-3M-IFN-β细胞还抑制了旁观者未转导的前列腺癌细胞的致瘤性。在用抗去唾液酸GM1抗体处理的裸鼠和严重联合免疫缺陷/米色小鼠中，PC-3M-IFN-β细胞产生了小肿瘤（直径3-5毫米）。免疫组织化学染色显示，PC-3M-IFN-β肿瘤被巨噬细胞均匀浸润，而对照肿瘤周边的巨噬细胞较少。对照肿瘤中的大多数肿瘤细胞用增殖细胞核抗原抗体染色呈阳性；很少有细胞用末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记呈阳性染色。形成鲜明对比的是，PC-3M-IFN-β肿瘤中增殖细胞核抗原阳性细胞较少，而末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记阳性细胞较多。用抗CD31抗体染色显示，对照肿瘤中的血管比PC-3M-IFN-β肿瘤中的血管多。与对照转导细胞相比，PC-3M-IFN-β细胞在体外对自然杀伤细胞介导的裂解或巨噬细胞介导的细胞生长抑制更敏感。来自PC-3M-IFN-β细胞的条件培养基增强了脾细胞介导的对对照肿瘤细胞的细胞溶解作用，这可以被抗IFN-β抗体中和。总体而言，数据表明PC-3M-IFN-β细胞致瘤性和转移的抑制是由于血管生成的抑制和宿主效应细胞的激活。
