中国癌症杂志
中國癌癥雜誌
중국암증잡지
CHINA ONCOLOGY
2014年
6期
401-405
,共5页
miR-22%胶质瘤%异粘蛋白%生长
miR-22%膠質瘤%異粘蛋白%生長
miR-22%효질류%이점단백%생장
miR-22%Glioma%MTDH%Growth
背景与目的:miR-22在胃癌、肺癌、结肠癌以及乳腺癌中表达下调,然而其在胶质瘤中的表达情况尚未明确。本研究旨在探讨miR-22是否通过靶向调控MTDH表达抑制胶质瘤细胞生长,从而进一步揭示miR-22的抑瘤机制。方法:运用实时定量PCR(quantitative real-time polymerase chain reaction,qRT-PCR)检测75例胶质瘤及17例正常大脑组织中miR-22的表达改变以及与胶质瘤患者预后关系;构建MTDH 3’UTR-荧光素酶报告载体,通过荧光素酶报告检测观察miR-22对MTDH 3’UTR-荧光素酶活性的影响;将miR-22 mimics转染胶质瘤细胞U251,以MTDH siRNA为阳性对照,采用蛋白质印迹法(Western blot)检测其对MTDH蛋白表达水平的影响;然后采用MTT法检测高表达miR-22对U251细胞生长的影响。结果:qRT-PCR检测结果显示,miR-22在75例胶质瘤组织中表达下调;Kaplan-Meier生存分析发现,miR-22表达越高,患者生存率越高(P<0.05);在双荧光素酶报告检测显示,miR-22能特异性地与MTDH 3’UTR结合,抑制其荧光素酶活性。Western blot检测结果显示,miR-22过表达或干扰MTDH可抑制MTDH蛋白的表达。MTT检测发现,miR-22过表达或干扰MTDH可抑制人U251细胞的生长,差异有统计学意义(P<0.05)。结论:miR-22通过靶向调控MTDH的表达而抑制胶质瘤细胞的生长。
揹景與目的:miR-22在胃癌、肺癌、結腸癌以及乳腺癌中錶達下調,然而其在膠質瘤中的錶達情況尚未明確。本研究旨在探討miR-22是否通過靶嚮調控MTDH錶達抑製膠質瘤細胞生長,從而進一步揭示miR-22的抑瘤機製。方法:運用實時定量PCR(quantitative real-time polymerase chain reaction,qRT-PCR)檢測75例膠質瘤及17例正常大腦組織中miR-22的錶達改變以及與膠質瘤患者預後關繫;構建MTDH 3’UTR-熒光素酶報告載體,通過熒光素酶報告檢測觀察miR-22對MTDH 3’UTR-熒光素酶活性的影響;將miR-22 mimics轉染膠質瘤細胞U251,以MTDH siRNA為暘性對照,採用蛋白質印跡法(Western blot)檢測其對MTDH蛋白錶達水平的影響;然後採用MTT法檢測高錶達miR-22對U251細胞生長的影響。結果:qRT-PCR檢測結果顯示,miR-22在75例膠質瘤組織中錶達下調;Kaplan-Meier生存分析髮現,miR-22錶達越高,患者生存率越高(P<0.05);在雙熒光素酶報告檢測顯示,miR-22能特異性地與MTDH 3’UTR結閤,抑製其熒光素酶活性。Western blot檢測結果顯示,miR-22過錶達或榦擾MTDH可抑製MTDH蛋白的錶達。MTT檢測髮現,miR-22過錶達或榦擾MTDH可抑製人U251細胞的生長,差異有統計學意義(P<0.05)。結論:miR-22通過靶嚮調控MTDH的錶達而抑製膠質瘤細胞的生長。
배경여목적:miR-22재위암、폐암、결장암이급유선암중표체하조,연이기재효질류중적표체정황상미명학。본연구지재탐토miR-22시부통과파향조공MTDH표체억제효질류세포생장,종이진일보게시miR-22적억류궤제。방법:운용실시정량PCR(quantitative real-time polymerase chain reaction,qRT-PCR)검측75례효질류급17례정상대뇌조직중miR-22적표체개변이급여효질류환자예후관계;구건MTDH 3’UTR-형광소매보고재체,통과형광소매보고검측관찰miR-22대MTDH 3’UTR-형광소매활성적영향;장miR-22 mimics전염효질류세포U251,이MTDH siRNA위양성대조,채용단백질인적법(Western blot)검측기대MTDH단백표체수평적영향;연후채용MTT법검측고표체miR-22대U251세포생장적영향。결과:qRT-PCR검측결과현시,miR-22재75례효질류조직중표체하조;Kaplan-Meier생존분석발현,miR-22표체월고,환자생존솔월고(P<0.05);재쌍형광소매보고검측현시,miR-22능특이성지여MTDH 3’UTR결합,억제기형광소매활성。Western blot검측결과현시,miR-22과표체혹간우MTDH가억제MTDH단백적표체。MTT검측발현,miR-22과표체혹간우MTDH가억제인U251세포적생장,차이유통계학의의(P<0.05)。결론:miR-22통과파향조공MTDH적표체이억제효질류세포적생장。
Background and purpose:miR-22 has been reported to be down-regulated in gastric cancer, lung cancer, colorectal cancer, and breast cancer. However, its expression in glioma was still poorly known. This study aimed to explicit whether miR-22 suppresses cell proliferation by targeting MTDH, thus to reveal molecular mechanism that miR-22 functions as a tumor suppressor in glioma. Methods:Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted for detecting the expression of miR-22 in gliomas and normal brain tissues. MTDH 3’UTR-luciferase vector was constructed and dual-luciferase reporter gene assay was employed to examine the effect of miR-22 on luciferase activity. U251 cells were transfected with miR-22 mimics, and MTDH siRNA as for postive control, then Western blot was performed to detect the expressions of MTDH protein. The proliferation ability of U251 cells was evaluated by MTT assay. Results:miR-22 was down-regulated in glioma tissues. Glioma patients with relatively high expression of miR-22 showed lower mortality compared with low expression of miR-22 by using Kaplan-Meier survival curves. We demonstrated miR-22 could bind to the 3’ untranslated region (UTR) of MTDH and inhibited the luciferase activity. Western blot showed that the expression of MTDH protein was inhibited by restored miR-22 or siR MTDH in U251 cells. Overexpression of miR-22 or siR MTDH inhibited the proliferation of U251 cells. Conclusion:miR-22 suppresses cell proliferation by targeting MTDH in glioma.