微小RNA-21对食管癌肌球家族蛋白l表达的抑制作用  

MicroRNA - 21 functionally suppresses the expression of tropomyosin 1 in esophageal cancer

作　　者：黎亮[1] 陈柏深[2] 沈卓坚[1] 甘向峰 徐夏[1] 陈炬[1] 

机构地区：[1]海南省人民医院胸外科,海口570105 [2]中山大学孙逸仙纪念医院胸外科

出　　处：《中华实验外科杂志》2015年第8期1814-1816,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨在食管癌中微小RNA-21（miRNA，miR-21）靶向蒯控肌球家族蛋白1（TPMI）表达的能力。方法对10例食管癌标本、癌旁上皮标本，分别采用定量聚合酶链反臆（（1PCR）和Westernblot检测miR-21（10例）和TPMl表达（6例）；在食管癌细胞株EC109中，采用荧光素酶报道基因实验鉴定miR-21靶向调控TPMI表达的能力；在ECl09中，利用小干扰RNA（siRNA）抑制miR-21，采用Westernblot检测TPM1的表达。结果miR-21在10例食管癌组织中的相对表达量为5．86±1．12，癌旁组织为1．56±0．32；TPM1在6例食管癌组织中的相对表达量为0．42±0．28，癌旁组织为6．58±2．32，miR-21与TPM1表达呈负相关。荧光素酶报道基因实验巾，导入奸生利TPMl后阴性对照组TPM1相对荧光度数为4．36±0．97，而miR-21组为3．06±0．37，两者比较贮译有统计学意义（P〈0．05）；而导人突变型TPM1后，两者比较差异无统计学意义（P〉0．05），说明miR-21靶向抑制TPMI表达。siRNA抑制miR-21后，TPM1从0升到5．68，TPMI表达恢复。结论n食管癌中，miR-21靶向抑制TPM1的表达，可能通过TPM1介导食管癌的侵袭与转移.Objective To determine the function of microRNA -21 （miRNA, miR -21 ） on reg- ulating expression of tropomyosin 1 （TPM1） in esophageal cancer. Methods Detected the expression of miR -21 （10 cases） and TPMI（6 cases） in 10 clinical esophageal cancer samples and adjacent normal epithelial samples via quantitative polymerase chain reaction （qPCR） and Western blot respectively; veri- fied the function of miR -21 on suppressing the expression of TPM1 in esophageal cancer via dual Luciferase Reporter Assay in esophageal cancer cell EC109 ; detected the expression of TPM1 after depression of miR-21 via anti -rniR -21 in esophageal cancer cell EC109. Results In clinical esophageal cancer samples and adjacent normal epithelial samples, the relative expression value of miR- 21 is 5.86 ± 1.12 and 1.56 ± 0. 32 respectively, while the relative expression value of TPM1 is 0. 42 ± 0. 28 and 6. 58± 2.32 respeetively. The result showed that the expression of miR -21 was negatively correlated with the expres- sion of TPMI. In dual luciferase reporter assay, the relative ]uciferase value decrease to 3.06 ± 0. 37 from 4.36 ± 0, 97 after inducing wild TPMI 3＇ UTR （ P 〈 0. 05 ）. Meanwhile, there is no statistically significant change after inducing mutated TPM1 3＇ UTR （ P 〈 0. 05 ） , showing that miR - 21 suppressed the expression of TPM1 directly in esophageal cancer cell EC109; Western blotting detection demonstrated that depression of miR - 21 could restore the expression of TPM1 from 0 to 5.68 in esophageal cancer cell ECI09. Conclusion In esophageal cancer, miR- 21 functionally suppresses the expression of TPMI , implicating that it maybe an important mechanism of miR - 21 inducing invasion and metastasis in esophageal cancer.

关 键 词：食管癌 微小RNA-21 肌球家族蛋白1 靶向 

分 类 号：R735.1[医药卫生—肿瘤]