摘要
目的探讨T细胞死亡相关基因8(TDAG8)通过AKT/mTOR参与胰腺癌疼痛作用的机制。方法将人胰腺癌细胞系SW 1990细胞移植到雌性BALB/c裸鼠胰腺中,建立胰腺癌致痛裸鼠模型。5周龄雌性BALB/C裸鼠48只随机分为假手术组(Sham组)、模型组(Model组)、NC-siRNA组、TDAG8-siRNA组,每组12只。Sham组注射培养基作为对照,NC-siRNA组及TDAG8-siRNA组分别将NC-siRNC、TDAG8-siRNA注射至胰腺癌痛模型裸鼠腹腔中。测量体重及苏木素伊红(HE)染色检测胰腺组织病理学变化。实时荧光定量PCR(RT-qPCR)检测脊髓背角组织中TDAG8的表达;检测各组裸鼠驼背评分及腹部皮肤机械痛阈值;Western blotting检测脊髓背角组织中TDAG8、降钙素基因相关肽(CGRP)、P物质(SP)、丝氨酸/苏氨酸激酶(AKT)、p-AKT、哺乳动物雷帕霉素靶蛋白(mTOR)、p-mTOR蛋白水平。AKT通路抑制剂LY294002作用胰腺癌痛模型裸鼠,检测各组裸鼠驼背评分以及腹部皮肤机械痛阈值;Western blotting检测脊髓背角组织中CGRP、SP、AKT、p-AKT、mTOR、p-mTOR蛋白水平。结果BALB/c裸鼠移植SW 1990细胞后,体重显著减轻,观察到胰腺癌细胞呈条索状排列密集,可见炎性细胞浸润,脊髓背角组织TDAG8 mRNA和蛋白水平显著升高,裸鼠驼背评分升高及机械痛阈值显著降低,脊髓背角组织CGRP、SP、p-AKT、p-mTOR蛋白水平显著升高(均P<0.05);TDAG8-siRNA和LY294002作用后裸鼠驼背评分降低及机械痛阈值均显著升高,脊髓背角组织CGRP、SP、p-AKT、p-mTOR蛋白水平显著降低(均P<0.05)。结论抑制TDAG8可减轻胰腺癌疼痛,其机制可能与抑制AKT/mTOR信号通路的激活有关。
Objective To investigate the role of the T-cell death associated gene 8(TDAG8)in pancreatic cancer pain.Methods SW 1990 cells were transplanted into the pancreas of female BALB/c nude mice to establish a pancreatic cancer pain mouse model.Weight measurement and hematoxylin eosin(HE)staining were used to detect the pathological changes of pancreatic tissue.TDAG8-siRNA was intraperitoneally injected into pancreatic cancer pain model mice,and real-time fluorescent quantitative PCR(RT-qPCR)was used to detect the expression of TDAG8 in the spinal dorsal horn tissue.The hunchback score and abdominal skin mechanical pain threshold of each group of mice were detected.Western blotting was used to detect TDAG8,calcitonin gene-related peptide(CGRP),substance P(SP),serine/threonine kinase(AKT),p-AKT,mammalian target of rapamycin(mTOR),and p-mTOR protein levels in spinal dorsal horn tissues.AKT pathway inhibitor LY294002 was applied to pancreatic cancer pain model mice,and humpback score and abdominal skin mechanical pain threshold of each group were detected.Western blotting was used to detect the protein levels of CGRP,SP,AKT,p-AKT,mTOR and p-mTOR in spinal dorsal horn tissues.Results In BALB/c nude mice transplanted with SW 1990 cells,body weight was significantly reduced,abnormal acinar cells were observed in pancreatic tissue,TDAG8 mRNA and protein levels were significantly increased in spinal dorsal horn tissue,hump score and mechanical pain threshold were significantly decreased in mice,the protein levels of CGRP,SP,p-AKT,and p-mTOR in the spinal dorsal horn tissue were significantly increased(P<0.05).After the treatment of TDAG8-siRNA and LY294002,the mice′s humpback score decreased and the mechanical pain threshold increased significantly,and the protein levels of CGRP,SP,p-AKT,and p-mTOR in the spinal dorsal horn tissue were significantly decreased(P<0.05).Conclusion Inhibition of TDAG8 can reduce the pain of pancreatic cancer,and its mechanism may be related to the inhibition of the activation of AKT/mTOR signali
作者
吴琼
张振武
王哲银
张娟
赵妍
WU Qiong;ZHANG Zhenwu;WANG Zheyin;ZHANG Juan;ZHAO Yan(Department of Pain,Shenzhen People's Hospital,Shenzhen 518020,Guangdong,China)
出处
《西部医学》
2022年第10期1420-1425,1431,共7页
Medical Journal of West China
基金
吴阶平医学基金会临床科研专项资助基金(320.6710.19093-27)。
