Llorián-Salvador María, Pevida Marta, Fernández-García María Teresa, Lastra Ana, Obaya Alvaro, Cal Santiago, Hidalgo Agustín, Menéndez Luis, Baamonde Ana
Laboratorio de Farmacología, Facultad de Medicina, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), Universidad de Oviedo, Asturias, Spain.
Prostate. 2015 Jan;75(1):70-83. doi: 10.1002/pros.22893. Epub 2014 Sep 27.
Pain due to bone metastases of prostatic origin is a relevant clinical issue. We study here the nociceptive responses obtained in mice receiving the intratibial inoculation of RM1 prostate cancer cells.
10(2) -10(5) RM1 cells were inoculated to C57BL/6 mice and tumor development was analysed histologically and with luciferase-expressing RM1 cells. Spinal astroglial (GFAP) or microglial (Iba-1) expression was assessed with immunohistochemical methods and hypernociception was measured by the unilateral hot plate, the paw pressure and the von Frey tests. The analgesic effect of morphine, zoledronic acid or the CCR2 antagonist RS504393 was measured. Levels of the chemokines CCL2, CCL3, and CCL5 were determined by ELISA.
The inoculation of 10(3) RM1 cells induced tumoral growth in bone with a mixed osteoclastic/osteoblastic pattern and evoked astroglial, but not microglial, activation in the spinal cord. Hyperalgesia and allodynia were already established four days after inoculation and dose-dependently inhibited by the s.c. administration of morphine (1-5 mg/kg) or zoledronic acid (1-3 mg/kg). CCL2 and CCL5, but not CCL3, were released by RM1 cells in culture whereas only an increased presence of CCL2 was found in bone tumor homogenates. The administration of the CCR2 antagonist RS504393 (0.3-3 mg/kg) inhibited RM1 induced thermal hyperalgesia without modifying mechanical allodynia.
The intratibial inoculation of RM1 cells in immunocompetent mice induces hypernociceptive responses and can be useful to perform studies of bone cancer induced pain related to androgen-independent prostate cancer. The antinociceptive role derived from the blockade of the CCR2 chemokine receptors is further envisaged.
前列腺来源的骨转移引起的疼痛是一个重要的临床问题。我们在此研究了接受RM1前列腺癌细胞胫骨内接种的小鼠的伤害性反应。
将10(2)-10(5)个RM1细胞接种到C57BL/6小鼠体内，并通过组织学和表达荧光素酶的RM1细胞分析肿瘤发展情况。用免疫组化方法评估脊髓星形胶质细胞（GFAP）或小胶质细胞（Iba-1）的表达，并用单侧热板法、 paw压力法和von Frey试验测量痛觉过敏。测量吗啡、唑来膦酸或CCR2拮抗剂RS504393的镇痛效果。通过ELISA测定趋化因子CCL2、CCL3和CCL5的水平。
接种10(3)个RM1细胞可诱导骨肿瘤生长，呈现破骨细胞/成骨细胞混合模式，并引起脊髓星形胶质细胞而非小胶质细胞的激活。接种后四天即出现痛觉过敏和异常性疼痛，皮下注射吗啡（1-5mg/kg）或唑来膦酸（1-3mg/kg）可剂量依赖性地抑制这些症状。RM1细胞在培养中释放CCL2和CCL5，但不释放CCL3，而在骨肿瘤匀浆中仅发现CCL2含量增加。给予CCR2拮抗剂RS504393（0.3-3mg/kg）可抑制RM1诱导的热痛觉过敏，而不改变机械性异常性疼痛。
在免疫活性小鼠中胫骨内接种RM1细胞可诱导伤害性反应，有助于进行与雄激素非依赖性前列腺癌相关的骨癌性疼痛研究。进一步设想了阻断CCR2趋化因子受体所产生的抗伤害感受作用。
