肺腺癌表达TRPM8蛋白的意义  

Significance of TRPM8 protein expression in lung adenocarcinoma

作　　者：马洪明[1] 黄远顺 黄妙灵 张苗[1] 陈洋 黄志宏[1] 崔淼 梁婉玲 刘升明[1] 蔡兴东[1] MA Hong-ming;HUANG Yuan-shun;HUANG Miao-ling;ZHANG Miao;CHEN Yang;HUANG Zhi-hong;CUI Miao;LIANG Wan-ling;LIU Sheng-ming;CAI Xing-dong(Department of Respiratory Medicine,The First Affiliated Hospital of Jinan University,Guangzhou 510630,China;College of Life Science and Technology,Jinan University,Guangzhou 510632,China)

机构地区：[1]暨南大学附属第一医院呼吸内科,广东广州510630 [2]暨南大学生命科学技术学院,广东广州510632

出　　处：《中国病理生理杂志》2020年第9期1680-1689,共10页Chinese Journal of Pathophysiology

基　　金：广东省公益研究与能力建设专项基金项目(No.2014A020212441);暨南大学科研培育与创新基金医学联合基金项目(No.21615475)。

摘　　要：目的:探讨肺腺癌表达瞬时受体电位阳离子通道M亚家族成员8(TRPM8)蛋白的意义。方法:收集112例肺腺癌患者的肿瘤组织标本,对其进行4~5年临床随访。免疫组织化学染色检测肿瘤组织中TRPM8蛋白的表达,分析TRPM8蛋白表达与肺腺癌患者临床特征及预后的关系。培养A549肺腺癌细胞,通过感染慢病毒上调细胞中TRPM8蛋白的表达,CCK-8法及集落形成实验检测细胞增殖能力,流式细胞术检测细胞周期及凋亡,划痕实验及Transwell实验观察细胞迁移和侵袭。培养H1299肺腺癌细胞,通过感染慢病毒上调细胞中TRPM8表达后,分别在免疫缺陷小鼠左右臀部皮下注射对应空载体细胞及上调TRPM8的细胞,分析TRPM8对移植瘤生长的影响。结果:肺腺癌组织存在TRPM8表达。TRPM8蛋白高表达的肺腺癌患者,其4~5年累积生存率高于TRPM8蛋白低表达的患者(P=0.017),且其肿瘤最大径小于低表达的患者(P=0.028)。上调A549细胞中TRPM8的表达后,细胞活力较空载体组和亲本细胞组显著减弱,形成的集落数显著减少,处于S期的细胞比例显著升高,迁移和侵袭能力显著下降(P<0.01)。与空载体对照细胞株比较,上调H1299细胞中TRPM8的表达后,其在免疫缺陷小鼠皮下的移植瘤生长速率显著下降,肿瘤重量显著降低(P<0.01)。结论:TPRM8在肺腺癌中具有抑癌作用,低表达TRPM8蛋白的肺腺癌患者预后不良。AIM:To investigate the significance of transient receptor potential cation channel subfamily M member 8(TRPM8)protein expression in lung adenocarcinoma.METHODS:The tumor samples from 112 cases of patients with lung adenocarcinoma were collected in our hospital,and 4~5 years of follow-up was conducted.The protein expression of TRPM8 was analyzed by immunohistochemical staining,and the correlations between the TRPM8 protein expression and the clinical characteristics including prognosis of the patients with lung adenocarcinoma were investigated.After TRPM8 protein expression was up-regulated in A549 lung adenocarcinoma cells by lentiviral infection,the proliferation of A549 cells was analyzed by CCK-8 assay and colony formation assay,the cell cycle and apoptosis were analyzed by flow cytometry,and the migration and invasion abilities of the cells were measured by scratch experiment and Transwell assay.The TRPM8 protein expression was stably up-regulated in H1299 cells by lentiviral infection,and then the left and right buttocks of the immunodeficient mice were subcutaneously injected with empty vector control cells and TRPM8-overexpressing cells,respectively.The effects of TRPM8 on the growth of H1299 cell-derived xenograft tumor in immunodeficient mice were evaluated.RESULTS:The 4~5-year survival rate in the patients with high TRPM8 protein expression was significantly higher than that in the patients with low expression of TRPM8 protein(P=0.017).The tumor maximum diameter in the patients with high TRPM8 protein expression was significantly smaller than that in the patients with low TRPM8 protein expression(P=0.028).The viability,the number of colonies and the migration and invasion abilities of TRPM8-overexpressing A549 cells were significantly decreased as compared with empty vector and parental cells(P<0.01).The results of flow cytometry analysis showed that the proportion of A549 cells at S stage was significantly in creased in TRPM8 overexpression group as compared with empty vector group(P<0.01).The growth r

关 键 词：肺腺癌 瞬时受体电位阳离子通道M亚家族成员8 细胞增殖 细胞侵袭 

分 类 号：R734.2[医药卫生—肿瘤] R730.23[医药卫生—临床医学]