scholarly journals The role of PKCγ subunit of rACC neurons in the development of bone cancer pain in rats

2019 ◽  
Author(s):  
Hao Feng ◽  
Zequn Feng ◽  
Meng She ◽  
Gongming Wang ◽  
Guanghui Cheng ◽  
...  
Keyword(s):  
Cancer Pain ◽  
Sham Group ◽  
Lentiviral Vector ◽  
Bone Cancer ◽  
Bone Cancer Pain ◽  
Vehicle Group ◽  
Marrow Cavity ◽  
Withdrawal Threshold ◽  
Naive Group ◽  
Bone Marrow Cavity

Abstract Background：To explore the role of PKCγ subunit of rACC neurons in the development of bone cancer pain in rats. Methods：40 adult female SD rats were divided into five groups: blank control group (Naive group), sham operation group (Sham group), bone cancer pain group (BCP group)，empty lentiviral vector group（Vehicle group） and PKCγ/shRNA recombinant lentiviral vector group (PKCγ group). 10μl MADB-106 rat mammary cancer cells suspension was inoculated into the tibia bone marrow cavity of rats in BCP group and Vehicle group similarly. 10μl saline was inoculated into the proximal tibia bone marrow cavity of rats in Sham group. In the PKCγ group, the rats were taken the same treatment as the BCP group, and then 10μl shRNA/PKCγ recombinant lentivirus was injected into the bilateral rACC on the 7th day. The mechanical withdrawal threshold and thermal withdrawal latency were measured every 3 days to assess the rat pain behavior. Immunohistochemistry, Western blotting technology and immunofluorescence were used to detect the expression of PKCγ subunits in rat rACC neurons after operation. Results: From the 3rd day after operation, the mechanical withdrawal thresholds in BCP group, Vehicle group and PKCγ group were significantly decreased than those in Naive group and Sham group (P<0.05). Compared with the BCP group and Vehicle group the mechanical withdrawal threshold in the PKCγ group increased significantly (P<0.05). On the 3rd postoperative day, the thermal withdrawal latency in BCP, Vehicle and PKCγ groups was significantly longer than those in Naive and Sham groups（P＜0.05). From the 14th postoperative day, the TWL in PKCγ group was shorter than that in BCP and Vehicle groups (P<0.05). Western blot analysis showed that the expression of PKCγ in rACC neurons on the 14th day after operation in rats of BCP and Vehicle groups were significantly higher than that in Naive group and Sham group. (P<0.05) However, in the PKCγ group, the expression of PKCγ in rACC neurons was significantly lower than that in BCP group (P<0.05). Conclusion: Up-regulation of PKC subunit of rACC neurons in bone cancer pain rats contributes to the development of pain sensitivity in bone cancer.

scholarly journals Up-regulation of PKCγ subunits of rACC neurons contributes to the development of pain sensitivity in bone cancer rats

2019 ◽  
Author(s):  
Meng She ◽  
Hao Feng ◽  
Zequn Feng ◽  
Gongming Wang ◽  
Guanghui Cheng ◽  
...  
Keyword(s):  
Cancer Pain ◽  
Western Blotting ◽  
Mechanical Allodynia ◽  
Sham Group ◽  
Lentiviral Vector ◽  
Thermal Hyperalgesia ◽  
Bone Cancer ◽  
Bone Cancer Pain ◽  
Vehicle Group ◽  
Vector Group

Abstract To explore the role of PKCγ subunits of rostral anterior cingulate cortex (rACC) neurons in the development of bone cancer pain in rats. Healthy female Sprague-Dawley rats were randomly divided into five groups: blank control group (naive group), sham operation group (sham group), bone cancer pain group (BCP group), BCP plus empty lentiviral vector group (vehicle group) and BCP plus PKCγ/shRNA recombinant lentiviral vector group (PKCγ group). The BCP group, vehicle group and PKCγ group received a 10 µl intra-tibial injection of MADB-106 rat mammary carcinoma cell suspension (4.6×10 8 cell/ml). In comparison, the sham group received a 10 µl intra-tibial injection of saline. The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were assessed on pre-operation day 0 (baseline) and days 3, 7, 14 and 21 after intra-tibial injection, respectively. To downregulate the PKCγ subunits of rACC neurons, the PKCγ group received a 10 µl bilateral rACC injection of shRNA/PKCγ recombinant lentivirus (1.25×10 9 TU/ml) on the day 7 after intra-tibial injection, whereas the vehicle group received an injection of the same dose of empty lentiviral vector. Western blotting, immunohistochemical and immunofluorescence analysis were performed to detect the different expression of PKCγ subunits in rACC neurons among these groups on postoperative days 7 or 21. No significant difference in the baseline of MWT and TWL was found among these five groups ( P > 0.05). However, compared with the naive group and sham group, the rats with bone cancer (BCP group, vehicle group and PKCγ group) demonstrated marked mechanical allodynia and thermal hyperalgesia that was evoked starting on postoperative day 7 following intra-tibial injection of carcinoma cells ( P < 0.05). Meanwhile, the western blotting analysis also confirmed that the expression of PKCγ in rACC neurons was significantly increased in the BCP model groups ( P < 0.05). However, from postoperative days 14-21, the injection of shRNA/PKCγ recombinant lentivirus in the PKCγ group alleviated mechanical allodynia and thermal hyperalgesia ( P < 0.05).The present study indicates that up-regulation of PKCγ subunits of rACC neurons in bone cancer pain rats contributes to the development of bone cancer pain.

scholarly journals Up-regulation of PKCγ subunits of rACC neurons contributes to the development of pain sensitivity in bone cancer rats

2019 ◽  
Author(s):  
Meng She ◽  
Hao Feng ◽  
Zequn Feng ◽  
Gongming Wang ◽  
Guanghui Cheng ◽  
...  
Keyword(s):  
Cancer Pain ◽  
Western Blotting ◽  
Mechanical Allodynia ◽  
Sham Group ◽  
Lentiviral Vector ◽  
Thermal Hyperalgesia ◽  
Bone Cancer ◽  
Bone Cancer Pain ◽  
Vehicle Group ◽  
Vector Group

Abstract To explore the role of PKCγ subunits of rostral anterior cingulate cortex (rACC) neurons in the development of bone cancer pain in rats. Healthy female Sprague-Dawley rats were randomly divided into five groups: blank control group (naive group), sham operation group (sham group), bone cancer pain group (BCP group), BCP plus empty lentiviral vector group (vehicle group) and BCP plus PKCγ/shRNA recombinant lentiviral vector group (PKCγ group). The BCP group, vehicle group and PKCγ group received a 10 µl intra-tibial injection of MADB-106 rat mammary carcinoma cell suspension (4.6×10 8 cell/ml). In comparison, the sham group received a 10 µl intra-tibial injection of saline. The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were assessed on pre-operation day 0 (baseline) and days 3, 7, 14 and 21 after intra-tibial injection, respectively. To downregulate the PKCγ subunits of rACC neurons, the PKCγ group received a 10 µl bilateral rACC injection of shRNA/PKCγ recombinant lentivirus (1.25×10 9 TU/ml) on the day 7 after intra-tibial injection, whereas the vehicle group received an injection of the same dose of empty lentiviral vector. Western blotting, immunohistochemical and immunofluorescence analysis were performed to detect the different expression of PKCγ subunits in rACC neurons among these groups on postoperative days 7 or 21. No significant difference in the baseline of MWT and TWL was found among these five groups ( P > 0.05). However, compared with the naive group and sham group, the rats with bone cancer (BCP group, vehicle group and PKCγ group) demonstrated marked mechanical allodynia and thermal hyperalgesia that was evoked starting on postoperative day 7 following intra-tibial injection of carcinoma cells ( P < 0.05). Meanwhile, the western blotting analysis also confirmed that the expression of PKCγ in rACC neurons was significantly increased in the BCP model groups ( P < 0.05). However, from postoperative days 14-21, the injection of shRNA/PKCγ recombinant lentivirus in the PKCγ group alleviated mechanical allodynia and thermal hyperalgesia ( P < 0.05).The present study indicates that up-regulation of PKCγ subunits of rACC neurons in bone cancer pain rats contributes to the development of bone cancer pain.

scholarly journals The role of PKCγ subunit of rACC neurons in the development of bone cancer pain in rats

2018 ◽  
Author(s):  
Hao Feng ◽  
Zequn Feng ◽  
Meng She ◽  
Gongming Wang ◽  
Guanghui Cheng ◽  
...  
Keyword(s):  
Cancer Pain ◽  
Sham Group ◽  
Bone Cancer ◽  
Bone Cancer Pain ◽  
Sham Operation ◽  
Withdrawal Threshold ◽  
Withdrawal Latency ◽  
Naive Group ◽  
Significant Difference

Abstract Background: To explore the role of PKCγ subunit of rACC neurons in the development of bone cancer pain in rats. Methods: 32 adult female SD rats were divided into four groups: blank control group (Naive group), sham operation group (Sham group), bone cancer pain group (BCP group) and PKCγ/shRNA recombinant lentiviral vector group (PKCγ group). 10μl MADB-106 rat mammary cancer cells suspension, 10μl saline was inoculated into the proximal tibia bone marrow cavity of rats in BCP group and Sham group respectively. In the PKCγ group, the rats were taken the same treatment as the BCP group, and then 10μl shRNA/NR2B recombinant lentivirus was injected into the bilateral rACC on the 7th day. The mechanical withdrawal threshold and thermal withdrawal latency were measured every 3 days to assess the rat pain behavior. Immunohistochemistry and Western blotting technology were used to detect the expression of PKCγ subunits in rat rACC neurons after operation. Results: There was no significant difference in the mechanical withdrawal threshold and thermal withdrawal latency between the four groups (P>0.05). From the 3rd day after operation, the mechanical withdrawal thresholds in BCP group and PKCγ group were significantly decreased than those in Naive group and Sham group (P<0.05). Compared with the BCP group, the mechanical withdrawal threshold in the PKCγ group increased significantly (P<0.05). On the 3rd postoperative day, the thermal withdrawal latency in BCP and PKCγ groups was significantly longer than those in Naive and Sham groups（P＜0.05). From the 10th postoperative day, the thermal withdrawal latency in PKCγ group was shorter than that in BCP group (P<0.05). Western blot analysis showed that the expression of PKCγ in rACC neurons on the 14th day after operation in rats of BCP group was significantly higher than that in Naive group and Sham group. (P<0.05) However, in the PKCγ group, the expression of PKCγ in rACC neurons was significantly lower than that in BCP group (P<0.05). Conclusion: Up-regulation of PKC subunit of rACC neurons in bone cancer pain rats may be involved in the development of pain sensitivity in bone cancer. Key words: Bone cancer pain; rat; rACC; PKCγ; gene silencing

scholarly journals The TRPA1 Channel Mediates Mechanical Allodynia and Thermal Hyperalgesia in a Rat Bone Cancer Pain Model

2021 ◽  
Vol 2 ◽  
Author(s):  
Qiangwei Liu ◽  
Long Feng ◽  
Xiujing Han ◽  
Weidong Zhang ◽  
Hong Zhang ◽  
...  
Keyword(s):  
Protein Expression ◽  
Cancer Pain ◽  
Mechanical Allodynia ◽  
Sham Group ◽  
Thermal Hyperalgesia ◽  
Bone Cancer ◽  
Bone Cancer Pain ◽  
Left Tibia ◽  
Withdrawal Threshold ◽  
Mrna And Protein Expression

Background: Bone cancer pain (BCP) significantly affects patient quality of life, results in great bodily and emotional pain, and creates difficulties in follow-up treatment and normal life. Transient receptor potential ankyrin 1 (TRPA1) is an essential transduction ion channel related to neuropathic and inflammatory pain. However, the role of TRPA1 in BCP remains poorly understood. This study aimed to explore the relationship between TRPA1 and BCP.Methods: A BCP model was induced by Walker256 cells to the left tibia. The sham group was induced by normal saline to the left tibia. Thereafter, pain behaviors and TRPA1 expression between the BCP group and the sham group were observed on the 14th day of modeling. The TRPA1 antagonist A967079 (10 mg/kg) was injected via tail vein. TRPA1 antisense oligodeoxynucleotide (AS-ODN, 5 nmol/10 μl) and missense oligodeoxynucleotide (MS-ODN, 5 nmol/10 μl) were intrathecally delivered via a mini-osmotic pump for 5 consecutive days to assess the effect of TRPA1 on BCP. Behavioral tests were assessed preoperatively and postoperatively. Real-time quantitative PCR and western blot analyses were used to measure TRPA1 levels among the different groups.Results: The BCP model was successfully established via X-ray and pathological sections at 14 days. Compared to the sham group, the BCP group was more sensitive to mechanical stimuli, cool stimuli and hot stimuli. Intravenously injected A967079 can relieve paw mechanical withdrawal threshold and paw withdrawal thermal latency in rats with BCP. Moreover, AS-ODN can relieve paw mechanical withdrawal threshold and paw withdrawal thermal latency in rats with BCP. Additionally, relative mRNA and protein expression of TRPA1 in the BCP group were much higher than those in the sham group (14.55 ± 1.97 vs. 1 ± 0.04, P &lt; 0.01). Compared to the BCP group, the relative mRNA and protein expression of TRPA1 in the BCP+AS-ODN group was reduced (14.55 ± 1.97 vs. 2.59 ± 0.34, P &lt; 0.01).Conclusions: The TRPA1 channel mediates mechanical allodynia and thermal hyperalgesia in a rat BCP model.

scholarly journals Amitriptyline influences the mechanical withdrawal threshold in bone cancer pain rats by regulating glutamate transporter GLAST

2019 ◽  
Vol 15 ◽  
pp. 174480691985583 ◽  
Author(s):  
Simeng Ma ◽  
Xiaochun Zheng ◽  
Ting Zheng ◽  
Fengyi Huang ◽  
Jundan Jiang ◽  
...  
Keyword(s):  
Cancer Pain ◽  
Glutamate Transporter ◽  
Bone Cancer ◽  
Bone Cancer Pain ◽  
Withdrawal Threshold

scholarly journals The circular RNA circSlc7a11 promotes bone cancer pain pathogenesis in rats by modulating LLC-WRC 256 cell proliferation and apoptosis

2021 ◽  
Author(s):  
Han-Wen Chen ◽  
Xiao-Xia Zhang ◽  
Zhu-Ding Peng ◽  
Zu-Min Xing ◽  
Yi-Wen Zhang ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cancer Pain ◽  
Expression Profiles ◽  
Bone Cancer ◽  
Circular Rna ◽  
Differentially Expressed ◽  
Bone Cancer Pain ◽  
Circular Rnas ◽  
Altered Expression ◽  
Proliferation And Apoptosis

AbstractTreatment of bone cancer pain (BCP) caused by bone metastasis in advanced cancers remains a challenge in clinical oncology, and the underlying mechanisms of BCP are poorly understood. This study aimed to investigate the pathogenic roles of circular RNAs (circRNAs) in regulating cancer cell proliferation and BCP development. Eight differentially expressed circRNAs in the rat spinal cord were validated by agarose gel electrophoresis and Sanger sequencing. Expression of circRNAs and mRNAs was detected by quantitative RT-PCR. MTS assay and flow cytometry were performed to analyze cell proliferation and apoptosis, respectively. Differentially expressed mRNA profiles were characterized by deep RNA sequencing, hierarchical clustering, and functional categorization. The interactions among circRNAs, microRNAs (miRNAs), and mRNAs were predicted using TargetScan. Additionally, western blot was performed to determine the protein levels of Pax8, Isg15, and Cxcl10. Multiple circRNAs were differentially expressed in the spinal cords of BCP model rats; of these, circSlc7a11 showed the greatest increase in expression. The overexpression of circSlc7a11 significantly promoted cell proliferation and repressed apoptosis of LLC-WRC 256 and UMR-106 cells, whereas circSlc7a11 silencing produced the opposite effects. Altered expression of circSlc7a11 also induced substantial changes in the mRNA expression profiles of LLC-WRC 256 cells; these changes were linked to multiple apoptotic processes and signaling pathways, such as the chemokine signaling pathway, and formed a complex circRNA/miRNA/mRNA network. Additionally, Pax8, Isg15, and Cxc110 protein level in LLC-WRC 256 cells was consistent with the mRNA results. The circRNA circSlc7a11 regulates rat BCP development by modulating LLC-WRC 256 cell proliferation and apoptosis through multiple-signaling mechanisms.

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