miR-154-5p对肺癌骨转移的影响研究

Effect of miR-154-5p on Bone Metastasis in Lung Cancer

目的探讨miR-154-5p对肺癌骨转移的影响。方法本实验时间为2021年9月—2022年2月。选取BALB/c-nu小鼠60只,采用随机数字表法将小鼠分为载体组和miR-154-5p组,每组30只。将pMSCV puro反转录病毒载体或过表达miR-154-5p的A549细胞分别注入载体组、miR-154-5p组小鼠左心室。注药第80天,载体组剩余6只存活,miR-154-5p组剩余21只存活。载体组取6只小鼠,miR-154-5p组随机取6只小鼠,用于后期数据分析。注药第0、20、40、60、80天采用缩爪阈值评估小鼠机械性疼痛程度,注药第80天进行X线检查以评价小鼠骨转移评分,注药第80天采用苏木精-伊红染色检测溶骨性病变面积。从注药开始每天监测两组小鼠生存情况和无骨转移生存情况,连续记录80 d。结果干预方法与时间在缩爪阈值上存在交互作用(P<0.05),干预方法、时间在缩爪阈值上主效应显著(P<0.05);注药第40、60、80天,miR-154-5p组缩爪阈值高于载体组(P<0.05)。注药第80天,miR-154-5p组骨转移评分低于载体组,溶骨性病变面积小于载体组(P<0.05)。载体组生存率为20%,miR-154-5p组生存率为70.0%。miR-154-5p组生存率高于载体组(P<0.05)。载体组中位无骨转移生存期为35 d,无骨转移生存率为86.7%;miR-154-5p组中位无骨转移生存期为76 d,无骨转移生存率为50.0%。miR-154-5p组无骨转移生存率高于载体组(P<0.05)。结论miR-154-5p上调可提高肺癌小鼠的缩爪阈值,抑制骨转移,并延长小鼠生存期和无骨转移生存期。

Objective To investigate the effect of miR-154-5p on bone metastasis in lung cancer.Methods Experimental time of this study was from September 2021 to February 2022.Sixty BALB/c-nu mice were selected and divided into carrier group and miR-154-5p group using a random number table method,with 30 mice in each group.The pMSCV puro retrovirus vector or A549 cells overexpressing miR-154-5p were injected into the left ventricle of mice in carrier group and miR-154-5p group,respectively.On the 80th day of drug injection,6 mice in the carrier group survived and 21 mice in the miR-154-5p group survived.Six mice were selected from the carrier group and six mice were randomly selected from the miR-154-5p group for later data analysis.On the 0th,20th,40th,60th and 80th day of drug injection,the degree of mechanical pain in mice was evaluated by paw withdrawal threshold;on the 80th day of drug injection,the bone metastasis score was evaluated by X-ray examination,and the osteolytic lesion area was detected by hematoxylin eosin staining.The survival and bone metastasis-free survival of mice were monitored every day for 80 days.Results There was an interaction between intervention methods and time on the paw withdrawal threshold(P<0.05),both intervention methods and time produced significant main effects on the paw withdrawal threshold(P<0.05).On the 40th,60th and 80th day of drug injection,the paw withdrawal threshold in miR-154-5p group was higher than that in carrier group(P<0.05).On the 80th day of drug injection,the bone metastasis score in miR-154-5p group was lower than that in carrier group,and osteolytic lesion area was smaller than that in carrier group(P<0.05).The overall survival rate of carrier group was 20.0%,the overall survival rate of miR-154-5p group was 70.0%.The survival rate in miR-154-5p group was higher than that in carrier group(P<0.05).The median bone metastasis-free survival time in the carrier group was 35 days,and the bone metastasis-free survival rate was 86.7%.The median bone metastasis-free survival time in the miR-154-5p group was 76 days,and the bone metastasis-free survival rate was 50.0%.The bone metastasis-free survival rate in miR-154-5p group was higher than that in carrier group(P<0.05).Conclusion Upregulation of miR-154-5p can increase the paw withdrawal threshold of lung cancer mice,reduce the bone metastasis,and prolong the survival time and bone metastasis-free survival time.