丁卡因通过MDM2基因下调Survivin蛋白抑制黑色素瘤生长  

Tetracaine inhibits melanoma growth by down-regulating Survivin through MDM2

作　　者：伍春林 易鹏 黄祥 冯建国 姜鲜 WU Chunlin;YI Peng;HUANG Xiang;FENG Jianguo;JIANG Xian(Department of Anesthesia,Affiliated Hospital of Southwest Medical University,Luzhou 646000,China;Department of Anesthesia,Luzhou People’s Hospital)

机构地区：[1]西南医科大学附属医院麻醉科,泸州646000 [2]泸州市人民医院麻醉科

出　　处：《山西医科大学学报》2021年第4期403-408,共6页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81902796);泸州市人民政府西南医科大学科技战略合作项目(2018LZXNYD-ZK05);高校大学生创新创业训练计划项目(S201816032157)。

摘　　要：目的研究局麻药丁卡因对黑色素瘤生长的影响及其可能机制。方法一系列梯度浓度的丁卡因(0.1-0.6 mmol/L)分别干预黑色素瘤细胞(A375和B16)24 h,MTT实验检测黑色素瘤的细胞活力,细胞集落实验检测黑色素瘤的增殖能力,Western blot检测细胞凋亡抑制蛋白Survivin及其上游基因MDM2表达的变化。取12只6-8周龄的BALB/c小鼠于背部建立皮下黑色素瘤模型,随机分为对照组(n=6)和丁卡因组(n=6)。丁卡因组经腹腔予以丁卡因处理,对照组经腹腔予以等量生理盐水。观察并记录肿瘤生长情况,处理结束后取肿瘤组织行免疫组化染色检测Survivin的表达变化。结果丁卡因可显著抑制黑色素瘤细胞A375和B16的细胞活力(P<0.001),且呈浓度依赖性,IC_(50)分别为0.41 mmol/L和0.16 mmol/L。与对照组相比,丁卡因组黑色素瘤细胞集落生长明显被抑制(P<0.001),Survivin和MDM2的表达明显下调(P<0.001)。在动物试验中,丁卡因组小鼠肿瘤体积及质量明显小于对照组(P<0.05),肿瘤组织免疫组化染色显示丁卡因组Survivin阳性表达相较于对照组表达减少(P<0.001)。结论局麻药丁卡因在细胞及动物水平上均能明显抑制黑色素瘤的生长,这可能与其通过MDM2下调Survivin的表达相关。Objective To investigate the effect of tetracaine on melanoma growth and its mechanism.Methods MTT assay was used to detect A375 and B16 cell viability after treated with tetracaine in a gradient concentration range of 0.1-0.6 mmol/L for 24 h.Colony formation assay was used to investigate the growth ability of melanoma.Western blot was used to investigate the expression of Survivin and its upstream gene MDM2.Twelve BALB/c mice(6-8 weeks old)were selected to establish a subcutaneous melanoma model on the back,and then they were randomly divided into control group(n=6)and tetracaine group(n=6).The mice were intraperito-neally injected with tetracaine in tetracaine group and the same volume of saline in control group.Subsequently,the growth of tumors was observed and recorded.Immunohistochemical staining was used to detect the expression of Survivin in melanoma tissues.Results Tetracaine inhibited the cell viability of melanoma cell lines A375 and B16 in a concentration-dependent manner(P<0.001),and the IC_(50)of tetracaine was respectively 0.41 mmol/L and 0.16 mmol/L.Compared with control group,the colony growth of melanoma was significantly inhibited in tetracaine group(P<0.001),and the Survivin and MDM2 expression levels were significantly down-regulated in A375 and B16(P<0.001).In animal experiments,the volume and the mass of tumor in tetracaine group were significantly smaller than those in control group(P<0.05).Immunohistochemical staining of tumor tissues showed that the expression of Survivin in tetracaine group was decreased compared with control group(P<0.001).Conclusion Tetracaine can significantly inhibit the melanoma growth at both cellular and animal experiments,which may be related to the down-regulation of Survivin expression by MDM2.

关 键 词：局麻药 丁卡因 黑色素瘤 SURVIVIN 

分 类 号：R739.5[医药卫生—肿瘤]