中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
8期
1814-1816
,共3页
黎亮%陈柏深%沈卓坚%甘向峰%徐夏%陈炬
黎亮%陳柏深%瀋卓堅%甘嚮峰%徐夏%陳炬
려량%진백심%침탁견%감향봉%서하%진거
食管癌%微小RNA-21%肌球家族蛋白1%靶向
食管癌%微小RNA-21%肌毬傢族蛋白1%靶嚮
식관암%미소RNA-21%기구가족단백1%파향
Esophageal cancer%MicroRNA-21%Tropomyosin 1%Target
目的 探讨在食管癌中微小RNA-21(miRNA,miR-21)靶向调控肌球家族蛋白1(TPM1)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应(qPCR)和Western blot检测miR-21(10例)和TPM1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TPM1表达的能力;在EC109中,利用小干扰RNA(siRNA)抑制miR-21,采用Western blot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中,导入野生型TPM1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±0.37,两者比较差异有统计学意义(P＜0.05);而导入突变型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TPM1表达.siRNA抑制miR-21后,TPM1从0升到5.68,TPM1表达恢复.结论 在食管癌中,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.
目的 探討在食管癌中微小RNA-21(miRNA,miR-21)靶嚮調控肌毬傢族蛋白1(TPM1)錶達的能力.方法 對10例食管癌標本、癌徬上皮標本,分彆採用定量聚閤酶鏈反應(qPCR)和Western blot檢測miR-21(10例)和TPM1錶達(6例);在食管癌細胞株EC109中,採用熒光素酶報道基因實驗鑒定miR-21靶嚮調控TPM1錶達的能力;在EC109中,利用小榦擾RNA(siRNA)抑製miR-21,採用Western blot檢測TPM1的錶達.結果 miR-21在10例食管癌組織中的相對錶達量為5.86 ±1.12,癌徬組織為1.56 ±0.32;TPM1在6例食管癌組織中的相對錶達量為0.42±0.28,癌徬組織為6.58±2.32,miR-21與TPM1錶達呈負相關.熒光素酶報道基因實驗中,導入野生型TPM1後陰性對照組TPM1相對熒光度數為4.36±0.97,而miR-21組為3.06±0.37,兩者比較差異有統計學意義(P＜0.05);而導入突變型TPM1後,兩者比較差異無統計學意義(P＞0.05),說明miR-21靶嚮抑製TPM1錶達.siRNA抑製miR-21後,TPM1從0升到5.68,TPM1錶達恢複.結論 在食管癌中,miR-21靶嚮抑製TPM1的錶達,可能通過TPM1介導食管癌的侵襲與轉移.
목적 탐토재식관암중미소RNA-21(miRNA,miR-21)파향조공기구가족단백1(TPM1)표체적능력.방법 대10례식관암표본、암방상피표본,분별채용정량취합매련반응(qPCR)화Western blot검측miR-21(10례)화TPM1표체(6례);재식관암세포주EC109중,채용형광소매보도기인실험감정miR-21파향조공TPM1표체적능력;재EC109중,이용소간우RNA(siRNA)억제miR-21,채용Western blot검측TPM1적표체.결과 miR-21재10례식관암조직중적상대표체량위5.86 ±1.12,암방조직위1.56 ±0.32;TPM1재6례식관암조직중적상대표체량위0.42±0.28,암방조직위6.58±2.32,miR-21여TPM1표체정부상관.형광소매보도기인실험중,도입야생형TPM1후음성대조조TPM1상대형광도수위4.36±0.97,이miR-21조위3.06±0.37,량자비교차이유통계학의의(P＜0.05);이도입돌변형TPM1후,량자비교차이무통계학의의(P＞0.05),설명miR-21파향억제TPM1표체.siRNA억제miR-21후,TPM1종0승도5.68,TPM1표체회복.결론 재식관암중,miR-21파향억제TPM1적표체,가능통과TPM1개도식관암적침습여전이.
Objective To determine the function of microRNA-21 (miRNA,miR-21) on regulating expression of tropomyosin 1 (TPM1) in esophageal cancer.Methods Detected the expression of miR-21 (10 cases) and TPM1 (6 cases) in 10 clinical esophageal cancer samples and adjacent normal epithelial samples via quantitative polymerase chain reaction (qPCR) and Western blot respectively;verified 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-miR-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 respectively.The result showed that the expression of miR-21 was negatively correlated with the expression of TPM1.In dual luciferase reporter assay,the relative luciferase value decrease to 3.06 ± 0.37 from 4.36 ± 0.97 after inducing wild TPM1 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 EC109.Conclusion In esophageal cancer,miR-21 functionally suppresses the expression of TPM1,implicating that it maybe an important mechanism of miR-21 inducing invasion and metastasis in esophageal cancer.
