MicroRNA-93-5p may participate in the formation of morphine tolerance in bone cancer pain mouse model by targeting Smad5
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Wen-Feng Xiao1,*, Yu-Sheng Li1,*, Wei Lou2, Ting Cai3, Shun Zhang4, Xiao-Ying Hu5, Xing-Wang Zhang6, Wei Luo1
1Department of Orthopaedics, Xiangya Hospital, Central South University, Changsha 410008, China
2Department of Pain Treatment, Ningbo No.2 Hospital, Ningbo 315010, China
3Department of Emergency Internal Medicine, Ningbo No.2 Hospital, Ningbo 315010, China
4Stem Cell Laboratory, Ningbo No.2 Hospital, Ningbo 315010, China
5Department of Nephropathy Dialysis Center, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
6Division of Pharmaceutics, College of Pharmacy, Jinan Unversity, Guangzhou 510632, P.R. China
*These authors have contributed equally to this work
Wei Luo, email: [email protected]
Ting Cai, email: [email protected]
Keywords: microRNA-93-5p, Smad5, bone cancer pain, morphine tolerance, paw mechanical withdrawal threshold
Received: February 29, 2016 Accepted: June 30, 2016 Published: July 11, 2016
Objective: In this study, we aim to find out the role of microRNA-93-5p (miR-93) and Smad5 in morphine tolerance in mouse models of bone cancer pain (BCP).
Results: At 7 days after injection of morphine, the PMWT showed no significant difference between the morphine model group and the saline model group (P < 0.05), suggesting that morphine tolerance had formed in the morphine model group. The morphine model group had higher miR-93 expression and lower Smad5 mRNA expression than the saline model group. Smad5 is a downstream target gene of miR-93. At 7, 9 and 14 days after injection of lentiviruses, the L/anti-miR-93 group had the lowest PMWTs, while the Smad5 shRNA group presented the highest PMWTs among these five groups (all P < 0.05).
Methods: We built mouse models of BCP and morphine tolerance and recorded 50% PMWT. After 6 days of modeling, we set saline control group, morphine control, saline model group and morphine model group (morphine tolerance emerged). We performed luciferase reporter gene assay to verify the relation between miR-93 and Smad5. After lentivirus transfection, the mice with morphine tolerance were assigned into L/anti-miR-93 group, Smad5 shRNA group, L/anti-miR-93 + Smad5 shRNA group, blank group and PBS control group. RT-qPCR, Western Blot assay and immumohistochemical staining were performed to observe the changes of miR-93 and Smad5.
Conclusion: Up-regulation of miR-93 may contribute to the progression of morphine tolerance by targeting Smad5 in mouse model of BCP.
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