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微小RNA-615-5p通过靶向热休克因子1调控乳腺癌细胞的增殖和凋亡。

MicroRNA-615-5p regulates the proliferation and apoptosis of breast cancer cells by targeting HSF1.

作者信息

Liu Kaisheng, Ma Rong

机构信息

Department of Breast Surgery, Qilu Hospital of Shandong University, Jinan, Shandong 250012, P.R. China.

出版信息

Exp Ther Med. 2021 Mar;21(3):192. doi: 10.3892/etm.2021.9624. Epub 2021 Jan 7.

Abstract

Breast cancer, which commonly occurs in the epithelium of the mammary gland, is a malignant tumor. MicroRNAs are involved in various cancer-associated processes, and microRNA-615-5p has been identified to be decreased in the pathological tissues from patients with breast cancer. In the present study, the possible mechanism of microRNA-615-5p in the progression of breast cancer was investigated in order to identify potential novel targets for clinical treatment. Heat shock factor 1 (HSF1) was identified as a predictive target gene of microRNA-615-5p using TargetScan analysis. The expression levels of microRNA-615-5p and its target gene, HSF1, were measured in breast cancer tissues and normal adjacent tissues. Additionally, the effects of microRNA-615-5p on MCF-7 breast cancer cell growth and apoptosis were examined. Furthermore, the interaction between HSF1 and microRNA-615-5p was investigated by a dual luciferase gene reporter assay. The expression levels of HSF1 were measured following transfection with microRNA-615-5p or pcDNA3.1-HSF1. Finally, the expression levels of proliferation- and apoptosis-associated factors such as B-cell lymphoma 2 (Bcl-2), cyclin D1, proliferating cell nuclear antigen (PCNA) and bcl-2-like protein 4 (Bax) were determined. The results demonstrated that lower microRNA-615-5p expression and higher HSF1 mRNA expression were present in tumor tissues compared with adjacent tissues (P<0.01). HSF1 was verified as a direct target of microRNA-615-5p using the dual luciferase gene reporter assay. In comparison with untransfected control and mimic-transfected negative control (NC) cells, MCF-7 cells transfected with microRNA-615-5p mimics exhibited reduced cell proliferation and increased apoptosis (P<0.01). However, the overexpression of HSF1 using a vector reversed the suppression of HSF1 induced by microRNA-615-5p mimics (P<0.01). The mRNA and protein expression levels of Bax were significantly increased, whereas those of Bcl-2, cyclin D1 and PCNA were decreased in the cells transfected with microRNA-615-5p mimics compared with the control and NC cells (P<0.01). Collectively, the present study indicated that microRNA-615-5p may mediate the progression of breast cancer by targeting HSF1.

摘要

乳腺癌是一种恶性肿瘤,常见于乳腺上皮。微小RNA参与多种癌症相关过程,且已发现在乳腺癌患者的病理组织中微小RNA - 615 - 5p水平降低。在本研究中,为了确定潜在的临床治疗新靶点,对微小RNA - 615 - 5p在乳腺癌进展中的可能机制进行了研究。使用TargetScan分析确定热休克因子1(HSF1)为微小RNA - 615 - 5p的预测靶基因。检测了乳腺癌组织和癌旁正常组织中微小RNA - 615 - 5p及其靶基因HSF1的表达水平。此外,研究了微小RNA - 615 - 5p对MCF - 7乳腺癌细胞生长和凋亡的影响。进一步通过双荧光素酶基因报告分析研究了HSF1与微小RNA - 615 - 5p之间的相互作用。在用微小RNA - 615 - 5p或pcDNA3.1 - HSF1转染后,检测了HSF1的表达水平。最后,测定了增殖和凋亡相关因子如B细胞淋巴瘤2(Bcl - 2)、细胞周期蛋白D1、增殖细胞核抗原(PCNA)和bcl - 2样蛋白4(Bax)的表达水平。结果表明,与癌旁组织相比,肿瘤组织中微小RNA - 615 - 5p表达较低,HSF1 mRNA表达较高(P<0.01)。使用双荧光素酶基因报告分析验证HSF1是微小RNA - 615 - 5p的直接靶标。与未转染的对照细胞和模拟物转染的阴性对照(NC)细胞相比,用微小RNA - 615 - 5p模拟物转染的MCF - 7细胞增殖减少,凋亡增加(P<0.01)。然而,使用载体过表达HSF1可逆转微小RNA - 615 - 5p模拟物诱导的HSF1抑制(P<0.01)。与对照细胞和NC细胞相比,用微小RNA - 615 - 5p模拟物转染的细胞中Bax的mRNA和蛋白表达水平显著升高,而Bcl - 2、细胞周期蛋白D1和PCNA的表达水平降低(P<0.01)。总体而言,本研究表明微小RNA - 615 - 5p可能通过靶向HSF1介导乳腺癌的进展。

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