Gm31083/miR-450b-5p/EphB1通路介导肺癌骨转移癌痛小鼠的机制研究

The mechanism underlying bone metastasis pain in lung cancer mediated by Gm31083/miR-450b-5p/EphB1 pathway

目的探讨Gm31083/miR-450b-5p/EphB1通路介导肺癌骨转移癌痛小鼠的机制。方法2020年1月至2021年12月,选取健康SPF级SD雄性小鼠20只,采用随机数字表法分为假手术组、模型组各10只。肺癌骨转移癌痛模型通过股骨内注射人肺腺癌A549细胞建立。于小鼠左侧足底中部采用2 g Von Frey细纤维刺激,5~6 s观察小鼠缩足反应;于热板仪上测定小鼠的痛阈值;采用实时荧光定量聚合酶链式反应法测定miR-450b-5p表达;采用Western Blot测定Gm31083和EphB1蛋白表达。结果模型组小鼠缩足反应率[(31.98±6.36)%]高于假手术组[(22.78±4.54)%](t=-3.72,P<0.05)。模型组小鼠热刺激痛阈值[(8.18±2.53)s]低于假手术组[(15.42±3.97)s](t=4.86,P<0.001)。模型组小鼠miR-450b-5p相对表达量(1.62±0.29)高于假手术组(1.00±0.04)(t=-6.70,P<0.001)。模型组小鼠Gm31083、EphB1蛋白表达灰度值分别为(1.23±0.21)、(2.73±0.28),均高于假手术组[(0.67±0.18)、(1.25±0.24)](t=-6.40、-12.69,均P<0.001)。结论Gm31083/miR-450b-5p/EphB1通路在肺癌骨转移癌痛小鼠中高表达,有可能成为评估肺癌骨转移癌痛新的标志物。

Objective To investigate the mechanism underlying bone metastasis pain in lung cancer mediated by Gm31083/miR-450b-5p/EphB1 pathway.Methods Between January 2020 and December 2021,a total of 20 healthy specific pathogen-free(SPF)grade Sprague-Dawley mice were selected and randomly divided into a sham operation group and a model group using a random number table method,with 10 mice in each group.A lung cancer bone metastasis pain model was established by injecting human lung adenocarcinoma A549 cells into the femur of each mouse in the model group.The middle part of the left plantar area of the mouse was stimulated with 2 g Von Frey fine fibers,and the mouse's foot retraction response was observed for 5-6 seconds.The pain threshold of each mouse was measured using a hot plate instrument.The expression of miR-450b-5p was determined by real-time fluorescence quantitative polymerase chain reaction.The expression of Gm31083 and EphB1 proteins was determined by western blot assay.Results The foot contraction response rate in the model group was(31.98±6.36)%,which was significantly higher than(22.78±4.54)%in the sham operation group(t=-3.72,P<0.05).The heat pain threshold in the model group was(8.18±2.53)seconds,which was significantly lower than(15.42±3.97)seconds in the sham operation group(t=4.86,P<0.001).The relative expression level of miR-450b-5p in the model group was(1.62±0.29),which was significantly higher than(1.00±0.04)in the sham operation group(t=-6.70,P<0.001).The grayscale values of Gm31083 and EphB1 proteins in the model group mice were(1.23±0.21)and(2.73±0.28),respectively,which were significantly higher than(0.67±0.18)and(1.25±0.24)in the sham operation group(t=-6.40,-12.69,both P<0.001).Conclusion Gm31083/miR-450b-5p/EphB1 is highly expressed in mice suffering from bone metastasis pain after developing lung cancer,and may become a new biomarker for evaluating bone metastasis pain in lung cancer.