Molecular mechanisms underlying the effect of intrathecal LV-SOCS3 injection on pain behaviors in a rat model of bone cancer
KONG Xiang-peng1, WANG Di-yü1, WEI Jin-rong1, YANG Zhi-xue1, LU Xiang-rong2, JIANG Guo-qin1
(1. Department of Surgery, the Second Affiliated Hospital, Soochow University, Suzhou Jiangsu 215004; 2. Department of Hand Surgery, the Affiliated Kunshan Hospital of Jiangsu University, Kunshan Jiangsu 215300, China)
Abstract:Objective: To investigate the effects of suppressor of cytokine signaling 3(SOCS3) overexpression by intrathecal injection lentiviral(LV)SOCS3 on the pain behaviors and potential mechanism in a rat model of bone cancer pain induced by breast cancer cells. Methods: Rats were injected with breast cancer cells to establish the models of bone cancer pain and control rats were injected with equal volume of normal saline (NS). Six model rats were treated by intrathecal LVSOCS3 injection, and the other six rats were treated by intrathecal injection of LVNC after one week of cancer cells injection.The paw withdrawal latency(PWL) to thermal stimulation and paw withdrawal threshold(PWT) to von Frey filament stimulation were carried out on control group, bone cancer group,LVSOCS3 group and LVNC group.Western blotting was performed to detect changes of SOCS3, NFκB and P2X3 receptors(P2X3R) at protein levels in L2-L5 dorsal root ganglion(DRGs) in different groups. Results: All rats presented enhanced mechanical and thermal hyperalgesia after intratibial injection of breast cancer cells, whose PWT and PWL were lower than those of control group. The protein level of SOCS3 was obviously reduced in bone cancer group.The PWT and PWL were higher than those of LVNC group at two weeks after intrathecal injection of LVSOCS3 but not at the time of one week after injection. SOCS3could be significantly increased and NFκBreduced in L2-L5DRGs at one week after intrathecal LVSOCS3 injection, but no change in P2X3R protein level was observed.At two weeks after intrathecal injection of LVSOCS3,the protein level of NFκBwas maintained at low level and P2X3R protein level was also significantly reduced. Conclusion: Intrathecal injection of LVSOCS3 could modulate the expression of NFκB and P2X3R in bone cancer pain rats, which resulted in attenuation of bone cancer pain.
孔祥鹏1, 王迪煜1, 魏金荣1, 杨志学1, 陆向荣2, 蒋国勤1. 鞘内注射慢病毒载体-SOCS3缓解骨癌痛大鼠痛行为及其机制[J]. 江苏大学学报:医学版, 2017, 27(02): 128-132.
KONG Xiang-peng1, WANG Di-yü1, WEI Jin-rong1, YANG Zhi-xue1, LU Xiang-rong2, JIANG Guo-qin1. Molecular mechanisms underlying the effect of intrathecal LV-SOCS3 injection on pain behaviors in a rat model of bone cancer. Journal of Jiangsu University(Medicine Edition), 2017, 27(02): 128-132.
[1\]Jemal A,Bray F, Center MM, et al. Global cancer statistics[J]. CA Cancer J Clin, 2015, 65(2):87-108.\[2\]Coleman RE.Clinical features of metastatic bone disease and risk of skeletal morbidity[J].Clin Cancer Res, 2006,12(20 Pt 2):6243s-6249s.\[3\]Liu X, Croker BA,Campbell IK, et al. Key role of suppressor of cytokine signaling 3 in regulating gp130 cytokineinduced signaling and limiting chondrocyte responses during murine inflammatory arthritis[J].Arthritis Reumatol,2014,66(9): 2391-2402.\[4\]Suzuki A, Hanada T, Mitsuyama K, et al. CIS3/SOCS3/SSI3 plays a negative regulatory role in STAT3 activation and intestinal inflammation[J].J Exp Med,2001, 193(4):471-481.\[5\]Dominguez E, Mauborgne A,Mallet J,et al.SOCS3mediated blockade of JAK/STAT3 signaling pathway reveals its major contribution to spinal cord neuroinflammation and mechanical allodynia after peripheral nerve injury[J].J Neurosci,2010, 30(16): 5754-5766.\[6\]Yi Y, Dang D, Chi TV, et al. Analgesia targeting IB4positive neurons in cancerinduced mechanical hypersensitivity[J].J Pain, 2012,13(6):524-531.\[7\]Xu GY, Huang LY. Peripheral inflammation sensitizes P2X receptormediated responses in rat dorsal root ganglion neurons[J].J Neurosci,2002,22(1):93-102.\[8\]Barclay J, Patel S, Dorn G, et al. Functional downregulation of P2X3 receptor subunit in rat sensory neurons reveals a significant role in chronic neuropathic and inflammatory pain[J]. J Neurosci,2002,22(22):8139-8147.\[9\]Hamilton SG, Mcmahon SB. ATP as a peripheral mediator of pain[J].J Auton Nerv Syst, 2000, 81(1/3):187-194.\[10\]Wu JX,Xu MY,Miao XR, et al. Functional upregulation of P2X3 receptors in dorsal root ganglion in a rat model of bone cancer pain[J]. Eur J Pain,2012,16(10):1378-1388.\[11\]Sun T,Luo J,Jia M,et al.Small interfering RNAmediated knockdown of NFκBp65 attenuates neuropathic pain following peripheral nerve injury in rats[J].Eur J Pharmacol,2012,682(1/3):79-85.\[12\]Wang C,Ning LP,Wang YH, et al. Nuclear factorκ B mediates TRPV4NO pathway involved in thermal hyperalgesia following chronic compression of the dorsal root ganglion in rats[J].Behav Brain Res,2011,221(1):19-24.\[13\]Fua ES, Yan PZ, Sagenb J, et al. Transgenic inhibition of glial NFκ B reduces pain behavior and inflammation after peripheral nerve injury[J].Pain,2010,148(3): 509-518.\[14\]Shi L, Zhang HH,Xiao Y, et al. Electroacupuncture suppresses mechanical allodynia and nuclear factor κ B signaling in streptozotocininduced diabetic rats[J].CNS Neurosci Ther,2013,19(2):83-90.\[15\]Zhang HH,Hu J,Zhou YL,et al.Promoted interaction of nuclear factorκB with demethylated cystathionineβsynthetase gene contributes to gastric hypersensitivity in diabetic rats[J]. J Neurosci,2013,33(21):9028-9038.\[16\]Zhou YL, Jiang GQ, Wei J, et al. Enhanced binding capability of nuclear factorкB with demethylated P2X3 receptor gene contributes to cancer pain in rats[J]. Pain, 2015, 156(10):1892-1905.\[17\]Zhang HH, Hu J,Zhou YL,et al.Promoted interaction of nuclear factorκB with demethylated purinergic P2X3 receptor gene contributes to neuropathic pain in rats with diabetes[J].Diabetes,2015,64(12):4272-4284.\[18\]Peters CM, Ghilardi JR, Keyser CP, et al.Tumorinduced injury of primary afferent sensory nerve fibers in bone cancer pain[J]. Exp Neurol,2005,193(1):85-100.\[19\]Schwei MJ, Honore P, Rogers SD, et al. Neurochemical and cellular reorganization of the spinal cord in a murine model of bone cancer pain[J].J Neurosci,1999,19(24): 10886-10897.\[20\]Ghilardi JR, Freeman KT, JimenezAndrade JM, et al. Administration of a tropomyosin receptor kinase inhibitor attenuates sarcomainduced nerve sprouting, neuroma formation and bone cancer pain[J]. Mol Pain, 2010, 6(1):1126-1132.\[21\]Zheng Q,Fang D, Liu M, et al. Suppression of KCNQM(Kv7) potassium channels in dorsal root ganglion neurons contributes to the development of bone cancer pain in a rat model[J].Pain,2013,154(3):434-448.\[22\]Larsen L,Rpke C.Suppressors of cytokine signalling:SOCS[J].APMIS,2002, 110(12): 833-844.\[23\]Trengove MC, Ward AC. SOCS proteins in development and disease[J]. Am J Clin Exp Immunol, 2013,2(1):1-29.\[24\]Souslova V, Cesare P, Ding Y, et al. Warmcoding deficits and aberrant inflammatory pain in mice lacking P2X3 receptors[J].Nature,2000,407(6807):1015-1017.\[25\]Li X, Li G, Xu H, et al. Effects of antirVEGF on the expression of VEGF receptor2 and P2X2/3 receptors of the spinal dorsal horn in neuropathic pain rats[J]. Brain Res Bull,2012,87(2/3):227-233.