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miR-204在小儿视网膜母细胞瘤中的表达及其对癌细胞增殖和凋亡的影响。

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

作者信息

Ding Jian, Lu Xiaoyun

机构信息

Medical Administration Division, The First Affiliated Hospital of Hunan Normal University (People's Hospital of Hunan Province), Changsha, Hunan 410005, P.R. China.

Department of Oncology, The First Affiliated Hospital of Hunan Normal University (People's Hospital of Hunan Province), Changsha, Hunan 410005, P.R. China.

出版信息

Oncol Lett. 2018 Dec;16(6):7152-7157. doi: 10.3892/ol.2018.9519. Epub 2018 Sep 27.

Abstract

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 µl 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 (p<0.001). The proportion of apoptotic cells in the experimental group was remarkably higher than that in the negative control group (p<0.001). 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 (p<0.001). 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.

摘要

研究了miR-204在人视网膜母细胞瘤(RB)及癌旁组织中的表达、临床意义及分子机制。收集了2013年4月至2017年6月在湖南师范大学第一附属医院(湖南省人民医院)眼科手术后保存的110例RB组织以及100例癌旁正常组织。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,并分析其与临床病理特征的相关性。用miR-204模拟物转染Y79细胞。将80 pmol/l的miR-204模拟物和10 μl Lipofectamine 2000加入实验组。在24、48、72和96小时通过甲基噻唑基四氮唑(MTT)法检测细胞增殖,在转染后48小时通过流式细胞术检测细胞凋亡,并通过RT-qPCR检测B细胞淋巴瘤2(Bcl-2)信使核糖核酸(mRNA)和Sirt1 mRNA的相对表达水平。MTT法结果显示,实验组在48小时的光密度(OD)测量值明显低于阴性对照组(p<0.001)。实验组凋亡细胞比例显著高于阴性对照组(p<0.001)。与阴性对照组相比,实验组中Bcl-2和Sirt1 mRNAs的相对表达水平显著降低(p<0.001)。miR-204可能参与RB的发生发展,这与患者的临床组织分化、神经浸润及淋巴结转移显著相关。miR-204可能通过下调RB中Bcl-2和Sirt1的表达来抑制RB细胞增殖并促进其凋亡。因此,miR-204可能成为RB早期诊断、预后评估及生物治疗的新生物学指标。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52fe/6256316/5f8299c7e497/ol-16-06-7152-g00.jpg

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