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. 2017 Jun 21;7:4005. doi: 10.1038/s41598-017-04198-3

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Intrathecal injection of CXCR4 siRNA attenuates TCI-induced cancer pain and up-regulation of p-CaMKII and p-CREB expression. Intrathecal injection of CXCR4 siRNA significantly attenuated TCI-induced mechanical allodynia (A and C), thermal hyperalgesia (B and D), spontaneous flinches (E) and limb uselessness (F) in bone cancer rats. CXCR4 siRNA (5 μg/10 μl, i.t.) or scrambled siRNA (negative control, 5 μg/10 μl, i.t.) was administered once daily on days 5, 6 and 7 (A and B) or days 12, 13 and 14 (C–F) after TCI. Behavioral tests were performed 12 hours after each injection. *P < 0.05, **P < 0.01 vs sham + scrambled siRNA group; ^# P < 0.05, ^## P < 0.01 vs TCI + scrambled siRNA group; n = 8 for each group. (G) Western blot analysis showing the inhibitory effect of CXCR4 siRNA on TCI-induced over-expression of p-CaMKII and p-CREB on day 14 after TCI. CXCR4 siRNA (siRNA, 5 μg/10 μl) or scrambled siRNA (Scr, 5 μg/10 μl) were administered once daily from days 12 to 14. L4-L6 spinal tissues were collected 12 hours after the last injection. **P < 0.01 vs sham group; ^# P < 0.05, ^## P < 0.01 vs TCI group; n = 4 for each group. (H) Immunofluorescence analysis showing inhibitory effect of CXCR4 siRNA on TCI-induced increase in mean fluorescence intensity (MFI) of p-CaMKII and the number (no.) of p-CREB-positive neurons on day 14 after TCI. CXCR4 siRNA (5 μg/10 μl) was administered once daily from days 12 to 14. L4-L6 spinal tissues were collected 12 hours after the last injection. **P < 0.01 vs sham group; ^# P < 0.05 vs TCI group; n = 4 for each group. Original magnification: 200×; scale bar: 100 μm.