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Expression of miR-204 in pediatric retinoblastoma and its effects on proliferation and apoptosis of cancer cells

机译:miR-204在儿科视网膜细胞瘤的表达及其对癌细胞增殖和凋亡的影响

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Expression, clinical significance and molecular mechanism of miR-204 in human retinoblastoma (RB) and para-carcinoma tissues were investigated. A total of 110 cases of RB tissues preserved after ophthalmectomy in the First Affiliated Hospital of Hunan Normal University (People's Hospital of Hunan Province) from April 2013 to June 2017 were collected along with 100 cases of para-carcinoma normal tissues. The expression of miR-204 in RB tissues was detected via reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and its associations with clinicopathological features were analyzed. Y79 cells were transfected with miR-204 mimics. A total of 80 pmol/l miR-204 mimics and 10 mu l Lipofectamine 2000 were added into the experimental group. Cell proliferation was detected via methyl thiazolyl tetrazolium (MTT) assay at 24, 48, 72 and 96 h, apoptosis was detected via flow cytometry at 48 h after transfection, and the relative expression levels of B-cell lymphoma 2 (Bcl-2) messenger RNA (mRNA) and Sirt1 mRNA were detected via RT-qPCR. The results of MTT assay revealed that the measured value of the optical density (OD) in the experimental group at 48 h was obviously lower than that in the negative control group (p0.001). The proportion of apoptotic cells in the experimental group was remarkably higher than that in the negative control group (p0.001). Compared with those in the negative control group, the relative expression levels of Bcl-2 and Sirt1 mRNAs in the experimental group were significantly decreased (p0.001). miR-204 may be involved in the occurrence and development of RB, which is significantly associated with clinical tissue differentiation, neural infiltration and lymph node metastasis in patients. miR-204 may inhibit proliferation and promote apoptosis of RB cells through downregulating the expression of Bcl-2 and Sirt1 in RB. Therefore, miR-204 may become a new biological index for early diagnosis, prognosis evaluation and biotherapy of RB.
机译:研究了MIR-204在人视网膜母细胞瘤(RB)和对癌组织中MIR-204的表达,临床意义和分子机制。从2013年4月到2017年4月到2017年4月,在湖南师范大学第一次附属医院(湖南省人民医院)后,共有110例RB组织患者。通过逆转录定量聚合酶链反应(RT-QPCR)检测RB组织中miR-204的表达,分析其与临床病理学特征的关联。用miR-204模拟转染Y79细胞。将总共​​80pmol / L miR-204模拟物和10μllipofectamine 2000加入到实验组中。通过在24,48,72和96h下通过甲基噻唑基四唑(MTT)测定检测细胞增殖,在转染后48小时内通过流式细胞术检测细胞凋亡,以及B细胞淋巴瘤2(BCL-2)的相对表达水平通过RT-QPCR检测信使RNA(mRNA)和SIRT1 mRNA。 MTT测定的结果显示,在48小时的实验组中的光密度(OD)的测量值明显低于阴性对照组(P <0.001)。实验组中凋亡细胞的比例显着高于阴性对照组(P <0.001)中的比例。与阴性对照组中的那些相比,实验组中Bcl-2和Sirt1 mRNA的相对表达水平显着降低(P <0.001)。 MIR-204可以参与RB的发生和发展,这与患者临床组织分化,神经浸润和淋巴结转移显着相关。 MiR-204可以通过下调Bcl-2和Sirt1在Rb中的表达来抑制增殖和促进RB细胞的凋亡。因此,MiR-204可能成为RB的早期诊断,预后评估和生物疗法的新生物指标。

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