The Kinesin Superfamily Protein 17 Contributes To The Development Of Bone Cancer Pain By Participating In NR2B Transport In The Spinal Cord Of Mice

Objective:To investigate the effects of the Kinesin superfamily protein 17(KIF17)contributes to the development of bone cancer pain by participating in the 2B subunit of N-methyl-D-aspartate receptor(NR2B)transport in the spinal cord of mice.Methods:Part One:Forty male C3H/HeJ mice,aged 4?6 weeks,weighting 20?25g,were randomly divided into two groups:sham operation group(group Sham,n=8)and bone cancer pain group(group Tumor,n=32).20 ?l ?-minimal essence medium(a-MEM)which containing 2×105 NCTC 2472 osteosarcoma cells was injected into the intramedullary space of the right femur in group Tumor.In group Sham,no cancer cell was instead.The number of spontaneous flinches(NSF)and the paw withdrawal mechanical threshold(PWMT)were measured on the day before(base)and 4 d,7 d,10 d,14 d after the operation.Then,the mice of group Tumor were randomly divided into four groups,KIF17 antisense ODN 2.5 ?g,5 ?g,10 ?g group(group A1,A2,A3)and the control group(group Ac).Group Sham and Ac were treated by saline.The pain behaviors were measured on 2 h,12 h,24 h,36 h and 48 h after administration.Part Two:sixty male C3H/HeJ mice were randomly divided into two groups:sham operation group(group S,n=20)and bone cancer pain group(group T,n=40).The group T underwent surgery for bone cancer pain.The pain behaviors and the protein expression of KIF17.mLin10 and NR2B in the spinal cord were measured on the day before(base)and 4 d,7 d,10 d,14 d after the operation.Then,the mice of group T were randomly divided into three groups:group TC,TS and TA.In group S and group TC,saline 5 ?l was injected intrathecally.In group TS and TA,5 ?g KIF17 sense ODN and 5 ?g antisense ODN,were injected for 6 consecutive days.Pain behaviors were assessed on 16 d,19 d,22 d,25 d,28 d,31 d after the operation.And tested the protein expression of KIF17,mLin10 and NR2B in the spinal cord on 19 d and 31d.Results:(1)Pain behaviors:Part One:Prior to the operation,there was no significant difference between groups in NSF or PWMT(P>0.05);Compared with group Sham,the NSF was increased and the PWMT was decreased on 7d,10 d and 14 d after operation in group Tumor(P<0.05);Compared with group Ac,the NSF was decreased and the PWMT was increased after administration in group A1,A2 and A3(P<0.05),However,on 36 h after administration in the group A1,48 h in the group A2 and A3,the pain behaviors were recovered to the level on 14 d.Part Two:The pain behaviors before dosing behavioural change in group S and T is consistent with the group Sham and group Tumor in part one;After administration,compared with group TC and TS,the NSF was decreased and the PWMT was increased significantly in the group TA(P<0.05),and the pain behaviors on 31 d after operation were recovered to the level on 14 d.(2)Western Blotting:Prior to the operation,there was no significant difference between group S and T;Compared with the group S,the protein expression of KIF17,mLin10,and NR2B in group T was increased on 7 d and peaked on 14 d after the operation(P<0.05);the protein expression levels of KIF17,mLin10 and NR2B in the spinal cord were significantly reduced following the application of KIF17 antisense ODN on 19 d after operation(P<0.05),On 31 d,the protein expression levels of KIF17,mLin10 and NR2B in the spinal cord were recovered to the level on 14 d.Conclusion:We determined that KIF17 played an important role in the process of bone cancer pain;Intrathecal administration of KIF17 antisense ODN attenuated the pain behaviors in the model of bone cancer pain and in a dose dependent of 2.5?5 ?g;The expression of KIF17,mLin10,NR2B were significantly increased in the spinal cord;The KIF17 antisense ODN attenuated pain behaviors through downregulating the expression of KIF17,mLin10,NR2B and inhibited the NR2B transport in the spinal cord of mice.