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时间:2020-05-03
《miR-148a对肝癌细胞株侵袭和迁移的抑制作用及机制-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、中国癌症杂志2014年第24卷第6期412CHINA0NCOLOGY2014Vo1.24No.6miR一148a对肝癌细胞株侵袭和迁移的抑制作用及机制贾筱琴缪俊俊雍军张子兰花晨李国利扬州大学医学院病理学教研室,江苏扬州225009【摘要]背景与目的:原发性肝癌是肝细胞或肝内胆管上皮发生的恶性肿瘤,其复发和转移十分常见。本实验以慢病毒介导miR-148a,感染人肝癌s删c一7721细胞,观察其对s~fMC~7721细胞侵袭和迁移能力的影响。方法:选用SMMC一7721肝癌细胞株,进行miR一148a慢病毒载体的构建,筛选稳定表达miR~148a肝癌细胞株。RT—PCR检测细胞中miR
2、~148a的表达水平。划痕实验及Transwell侵袭实验检测细胞侵袭迁移能力。采用明胶酶谱法测定MMP一2、ICNP一9的活性。蛋白质印迹法(Westernblot)检测MMP一2、MMP~9~1EMT相关蛋白(E-cadherin、vimentin)的表达情况。结果:RT-PCR结果显示,和对照组相比,LV—miR一148a肝癌细胞感染组表达miR一148a明显增高,体外侵袭实验细胞划痕实验显示过表达miR~148a后SMMC一7721的侵袭和迁移能力明显下降。明胶酶谱法结果显示过表达miR一148a的肝癌细胞,MMP一2和MMP~9降解明胶的能力下降(P3、iR~148a同时能降低肝癌细胞SMMC-7721中vimentin、MMP一2和MMP一9蛋白的表达,但并未影响E—cadherin的表达。结论:miR一148a在体外能抑制肝癌细胞SMMC一7721的侵袭和迁移,其机制可能与下调MMP一2、MMP一95~vimentin的表达水平有关。[关键词]miR-148a;肝细胞癌;侵袭;迁移;MMP一2;MMP一9DOI:10.3969/j.issn.1007—3969.2014.06.003中图分类号:R735.7文献标志码:A文章编号:1007—3639(2014)06—0412—06Inhibitionoftheinvasiona4、ndmigrationOfhepatOcellularcarcinomacellsbymiR-148aandthemechanismsJIAXiao—qin,MIAOJun-jun,YONGJun,ZHANGZi—lan,HUAChen,LIGuo—li(DepartmentofPathology,MedicalCollegeofYangzhouUniversity,YangzhouJiangsu225009,China)Correspondenceto:LIGuo—liE.mail:miaoyzu@sina.corn[Abstract]Backgroundandpurpose:P5、rimarylivercanceristhemalignanttumoroflivercellsorintrahepaticbileductepitheliumwithfamiliarmetastasisandpostsurgicalrecurrence.ThepurposeofthisstudywastoinvestigatetheefectsofmiR.148aontheinvasionandmigrationofhepatocellularcarcinomacellsandtheunderlyingmechanisms.Methods:Thesupernatantcontai6、ningLV_miR.148alentivirusparticleswasusedtoinfectSMMC一7721cells.TheexpressionofmiR.148awasdeterminedbyr-PCR.WoundhealingassayandtranswellassaywereperformedtodetecttheefectsofmiR.148aontheinvasionofhepatocellularcarcinomacells.GelatinzymographyassaywasusedtodetecttheeffectsofmiR一148aontheenzyme7、activitiesofmatrixmetanoDroteinase.2(MMP一21andmatrixmetalloproteinase.9(MMP.9).TheexpressionofMMP-2,MMP一9,E—cadherinandvimentinproteinswasdetectedbvWsternblotassay.Results:RT.PCRshowedtheexpressionofmiR一148awasupregulatedintheinfectedSM
3、iR~148a同时能降低肝癌细胞SMMC-7721中vimentin、MMP一2和MMP一9蛋白的表达,但并未影响E—cadherin的表达。结论:miR一148a在体外能抑制肝癌细胞SMMC一7721的侵袭和迁移,其机制可能与下调MMP一2、MMP一95~vimentin的表达水平有关。[关键词]miR-148a;肝细胞癌;侵袭;迁移;MMP一2;MMP一9DOI:10.3969/j.issn.1007—3969.2014.06.003中图分类号:R735.7文献标志码:A文章编号:1007—3639(2014)06—0412—06Inhibitionoftheinvasiona
4、ndmigrationOfhepatOcellularcarcinomacellsbymiR-148aandthemechanismsJIAXiao—qin,MIAOJun-jun,YONGJun,ZHANGZi—lan,HUAChen,LIGuo—li(DepartmentofPathology,MedicalCollegeofYangzhouUniversity,YangzhouJiangsu225009,China)Correspondenceto:LIGuo—liE.mail:miaoyzu@sina.corn[Abstract]Backgroundandpurpose:P
5、rimarylivercanceristhemalignanttumoroflivercellsorintrahepaticbileductepitheliumwithfamiliarmetastasisandpostsurgicalrecurrence.ThepurposeofthisstudywastoinvestigatetheefectsofmiR.148aontheinvasionandmigrationofhepatocellularcarcinomacellsandtheunderlyingmechanisms.Methods:Thesupernatantcontai
6、ningLV_miR.148alentivirusparticleswasusedtoinfectSMMC一7721cells.TheexpressionofmiR.148awasdeterminedbyr-PCR.WoundhealingassayandtranswellassaywereperformedtodetecttheefectsofmiR.148aontheinvasionofhepatocellularcarcinomacells.GelatinzymographyassaywasusedtodetecttheeffectsofmiR一148aontheenzyme
7、activitiesofmatrixmetanoDroteinase.2(MMP一21andmatrixmetalloproteinase.9(MMP.9).TheexpressionofMMP-2,MMP一9,E—cadherinandvimentinproteinswasdetectedbvWsternblotassay.Results:RT.PCRshowedtheexpressionofmiR一148awasupregulatedintheinfectedSM
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