miR-34b/c-5p 和神经激肽-1 受体调节乳腺癌细胞的增殖和凋亡。
MiR-34b/c-5p and the neurokinin-1 receptor regulate breast cancer cell proliferation and apoptosis.
机构信息
Department of Clinical Laboratory, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin's Clinical Research Center for Cancer, Tianjin Key Laboratory of Cancer Prevention and Therapy, Key Laboratory of Breast Cancer Prevention and Therapy of Educational Ministry, Tianjin Medical University, Tianjin, China.
Department of Clinical Laboratory, Aviation General Hospital, Beijing, China.
出版信息
Cell Prolif. 2019 Jan;52(1):e12527. doi: 10.1111/cpr.12527. Epub 2018 Oct 17.
OBJECTIVES
MiR-34 is a tumour suppressor in breast cancer. Neurokinin-1 receptor (NK1R), which is the predicted target of the miR-34 family, is overexpressed in many cancers. This study investigated the correlation and clinical significance of miR-34 and NK1R in breast cancer.
MATERIALS AND METHODS
Western blotting, quantitative reverse transcription-PCR (qRT-PCR) and luciferase assays were conducted to analyse the regulation of NK1R by miR-34 in MDA-MB-231, MCF-7, T47D, SK-BR-3 and HEK-293 T cells. MiR-34b/c-5p, full-length NK1R (NK1R-FL) and truncated NK1R (NK1R-Tr) expression in fifty patients were quantified by qRT-PCR and correlated with their clinicopathological parameters. CCK-8 assays, colony formation assays and flow cytometry were used to measure cell proliferation and apoptosis in MDA-MB-231 and MCF-7 cells transfected with miR-34b/c-5p or NK1R-siRNA and before treatment with or without Substance P (SP), an endogenous peptide agonists of NK1R. The effect of NK1R antagonist aprepitant was also investigated. In vivo xenograft models were used to further verify the regulation of NK1R by miR-34b/c-5p.
RESULTS
Expression levels of miR-34b/c-5p and NK1R-Tr, but not NK1R-FL, were associated with enhanced malignant potential, such as tumour stage and Ki67 expression. The overexpression of miR-34b/c-5p or NK1R silencing potently suppressed cell proliferation and induced G2/M phase arrest and the apoptosis of MDA-MB-231 and MCF-7 cells. The NK1R antagonist aprepitant had similar effects. In vivo studies confirmed that miR-34b/c-5p overexpression or NK1R silencing reduced the tumorigenicity of breast cancer. In addition, SP rescued the effects of miR-34b/c-5p overexpression or NK1R silencing on cell proliferation and apoptosis in vitro and in vivo assays.
CONCLUSIONS
MiR-34b/c-5p and NK1R contribute to breast cancer cell proliferation and apoptosis and are potential targets for breast cancer therapeutics.
目的
miR-34 是乳腺癌的肿瘤抑制因子。神经激肽-1 受体（NK1R）是 miR-34 家族的预测靶点，在许多癌症中过度表达。本研究探讨了 miR-34 与乳腺癌中 NK1R 的相关性及其临床意义。
材料与方法
采用 Western blot、定量逆转录-PCR（qRT-PCR）和荧光素酶报告基因实验分析 miR-34 在 MDA-MB-231、MCF-7、T47D、SK-BR-3 和 HEK-293T 细胞中对 NK1R 的调控作用。采用 qRT-PCR 检测 50 例患者的 miR-34b/c-5p、全长 NK1R（NK1R-FL）和截断型 NK1R（NK1R-Tr）的表达，并与临床病理参数相关分析。采用 CCK-8 检测、集落形成实验和流式细胞术检测转染 miR-34b/c-5p 或 NK1R-siRNA 后，以及用或不用 NK1R 的内源性肽激动剂 P 物质（SP）处理前后 MDA-MB-231 和 MCF-7 细胞的增殖和凋亡。还研究了 NK1R 拮抗剂 aprepitant 的作用。体内异种移植模型进一步验证了 miR-34b/c-5p 对 NK1R 的调控作用。
结果
miR-34b/c-5p 和 NK1R-Tr 的表达水平与恶性潜能增强相关，如肿瘤分期和 Ki67 表达。miR-34b/c-5p 的过表达或 NK1R 沉默可强烈抑制 MDA-MB-231 和 MCF-7 细胞的增殖，并诱导 G2/M 期阻滞和细胞凋亡。NK1R 拮抗剂 aprepitant 也有类似的作用。体内研究证实，miR-34b/c-5p 过表达或 NK1R 沉默可降低乳腺癌的致瘤性。此外，SP 挽救了 miR-34b/c-5p 过表达或 NK1R 沉默对体外和体内细胞增殖和凋亡的影响。
结论
miR-34b/c-5p 和 NK1R 促进乳腺癌细胞增殖和凋亡，是乳腺癌治疗的潜在靶点。
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