【摘要】 目的探讨微RNA(miRNA)-2l在大肠癌(CRC)细胞中的表达,抑制miRNA-21(miR-21)表达对大肠癌细胞中磷酸醋酶张力蛋白同源物(PTEN)、PI3K、蛋白激酶B(Akt)、基质金属蛋白酶-9(MMP-9)、mTOR表达的影响及其可能调控的靶基因。方法通过反义寡核苷酸(ASO)技术抑制大肠癌细胞株HCT116中miR-21的表达并加以验证。通过荧光定量PCR(FQ-PCR)技术及Western印迹法检测HCT116细胞中PTEN、PI3K、Akt、MMP-9、mTOR表达的改变。利用生物信息学分析预测miRNA-21可能的靶基因,荧光素酶双报告基因系统检测PTEN活性。结果 miR-21在HCT116中表达最高,在SW480中表达最低。miRNA-21在细胞株HCT116中的表达得到有效抑制,并且PTEN蛋白的表达明显上调,PI3K、Akt、MMP-9、mTOR表达下调。筛选并确认了PTEN为miR-21的调控靶基因之一。结论抑制miR-21的表达可促进HCT116细胞PTEN表达升高,PI3K、Akt、MMP-9、mTOR表达下调,PTEN可作为miR-21的靶基因参与调控过程。miRNA-21在CRC中表达失控,通过抑制PTEN表达促进肿瘤细胞侵袭转移。更多还原

【Abstract】 Objective To explore the expression of miRNA-21( miR-21) in colorectal cancer( CRC),discuss the potential role of miR-21 playing on target mRNA regulation. Methods The ASO technology was employed to suppress the expression of miR-21 in HCT116 cells. The changes in PTEN,PI3 K,Akt,MMP-9,mTOR mRNA and protein expressions were detected by FQ-PCR and Western blot respectively. The potential target mRNA prediction was performed by Bioinformatics. Dual report gene assay was used to identify the target gene of miR-21. Results The level of miR-21 was the highest in HCT116 cell,lowest in SW480 cell. The suppression of miR-21 could lead to the increment of PTEN protein,the down-regulation of PI3 K,Akt,MMP-9 and mTOR. PTEN was identified as one of target genes of miR-21 in CRC cell. Conclusions miR-21 is over-expressed in CRC and can promot cancer cell invasion by inhibiting the expression level of PTEN.更多还原