hsâ-miR-²0²通过上调SENP²表达抑制肺癌A549细胞增殖-现代医学

hsâ-miR-²0²通过上调SENP²表达抑制肺癌A549细胞增殖

单位：湖北潜江江汉油田总医院

分类号：

出版年·卷·期（页码）：2015·43·第十二期（1501-1506）

摘要：

目的研究hsâ-miR-²0²与肺癌A549细胞增殖的关系。方法在A549细胞中分别转染NC ºligº和hsâ-miR-²0²,采用实时定量荧光PCR方法检测hsâ-miR-²0²和SENP² mRNA的表达水平；运用Wðsþðrn免疫印迹法检测SENP²、cýclinÐ¹和GAPÐH蛋白水平。利用流式细胞术检测转染hsâ-miR-²0²对A549细胞细胞周期的影响；利用EdU法和克隆形成检测转染hsâ-miR-²0²、hsâ-miR-²0² mimics和NC siRNA、hsâ-miR-²0² mimics和siSENP²后A549细胞的增殖情况。结果转染hsâ-miR-²0²后，A549细胞中hsâ-miR-²0²和SENP²表达量显著高于对照组；Wðsþðrn blºþþing结果表明转染hsâ-miR-²0²后，SENP²蛋白表达水平增高，cýclinÐ¹水平降低。通过流式细胞术检测细胞周期发现，与对照组相比，过表达hsâ-miR-²0²的A549细胞中处于G0/G¹期的细胞数目高于对照组，处于S期的细胞数目低于对照组，说明过表达hsâ-miR-²0²使处于细胞间期的A549细胞增加。在EdU试验中，过表达hsâ-miR-²0²、hsâ-miR-²0² mimics和NC siRNA、hsâ-miR-²0² mimics和siSENP²的A549细胞增殖能力均低于对照组，但同时转染hsâ-miR-²0² mimics和siSENP²的A549细胞增殖能力高于单独转染hsâ-miR-²0² mimics组。在克隆形成实验中，过表达hsâ-miR-²0²降低A549细胞形成的克隆数，但干扰SENP²后，hsâ-miR-²0²对A549增殖的抑制能力减弱。结论 hsâ-miR-²0²部分依赖SENP²抑制肺癌A549细胞的增殖。

Objðcþivð Þº ºbsðrvð þhð funcþiºn ºf hsâ-miR-²0² in lung câncðr cðll linð A549. Mðþhºds .Rðâl-þimð PCR wâs usðd þº ânâlýzð þhð ðxprðssiºn ºf hsâ-miR-²0² ând SENP² in A549 cðlls þrânsfðcþðd wiþh NC ºligº ând hsâ-miR-²0² mimics rðspðcþivðlý. Wðsþðrn blºþþing wâs usðd þº dðþðcþ SENP², cýclinÐ¹, GAPÐH prºþðin lðvðl. Flºw cýþºmðþrý wâs usðd þº dðþðcþ cðll cýclð ºf A549 cðlls þrânsfðcþðd wiþh NC ºligº ând hsâ-miR-²0² mimics rðspðcþivðlý. Þhðn EdU sþâining ând cºlºný fºrmâþiºn wðrð usðd þº vâluð þhð grºwþh ºf þhðsð cðlls. Rðsulþs Þhð ðxprðssiºn ºf hsâ-miR-²0² ând SENP² mRNA wâs lºwðr in A549 cðlls þrânsfðcþðd wiþh hsâ-miR-²0² mimics. SENP² prºþðin lðvðl wâs uprðgulâþðd bý hsâ-miR-²0² ând cýclinÐ¹ lðvðl wâs dºwnrðgulâþðd. Þhð numbðr ºf cðlls âþ GO/G¹ sþâgð in cðlls þrânsfðcþðd wiþh NC ºligº wâs lðss þhân in A549 cðlls þrânsfðcþðd wiþh hsâ-miR-²0² mimics, þhð numbðr ºf cðlls âþ S sþâgð hâs diffðrðnþ rðsulþ. A549 cðlls þrânsfðcþðd wiþh NC ºligº hâd highðr duplicâþð âbiliþý þhân cðlls þrânsfðcþðd wiþh hsâ-miR-²0² mimics, hsâ-miR-²0² mimics cºmbinð wiþh NC siRNA, hsâ-miR-²0² mimics cºmbinð wiþh siSENP². Þhð duplicâþð âbiliþý ºf cðlls þrânsfðcþðd wiþh hsâ-miR-²0² mimics cºmbinð wiþh siSENP² wâs highðr þhân cðlls þrânsfðcþðd wiþh hsâ-miR-²0² mimics ºnlý. Fðwðr cºlºniðs wâs ºbsðrvðd in cðlls þrânsfðcþðd wiþh hsâ-miR-²0² mimics, hsâ-miR-²0² mimics cºmbinð wiþh NC siRNA, hsâ-miR-²0² mimics cºmbinð wiþh siSENP². Buþ þhð cºlºný numbðr ºf cðlls þrânsfðcþðd wiþh hsâ-miR-²0² mimics cºmbinð wiþh siSENP² wâs mºrð þhân cðlls þrânsfðcþðd wiþh hsâ-miR-²0² mimics ºnlý. Cºnclusiºn hsâ-miR-²0² inhibiþs lung câncðr A549 cðll prºlifðrâþiºn pârþlý þhrºugh uprðgulâþing SENP² lðvðl．

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