【摘要】 目的探讨microRNA-34a(mi R-34a)对鼻咽癌鳞状肿瘤干细胞表型的体外调控作用。方法利用原代培养的人类鼻咽癌细胞,采用流式分选法定量并收集ALDH(aldehyde dehydrogenase)+肿瘤干细胞。通过RT-qPCR分析mi R-34与肿瘤干细胞相关因子(Sox2、Nanog、Oct3/4)在ALDH~+和ALDH~-细胞中的表达情况。进一步用miR-34a拟态转染评估其对鼻咽癌肿瘤干细胞标记物以及相关因子的表达调控能力。结果原代鼻咽癌细胞中表达ALDH的细胞约占15.43%左右。与ALDH~-细胞相比,miR-34a表达水平在大多数ALDH~+细胞中显著下调(-13.2 vs-4.1倍),3种肿瘤干细胞相关因子表达显著增加(最高36.8倍)。ALDH~+细胞转染miR-34a拟态24 h、48 h、72 h后miR-34a mRNA水平显著增加,48 h时达峰值水平(4.49倍,P<0.01),而3种肿瘤干细胞相关因子表达水平显著降低。结论恢复miR-34a表达显著抑制人类原代鼻咽癌干细胞表型的形成。调控鼻咽癌和肿瘤干细胞中miR-34a的表达可能降低肿瘤治疗后的转移和复发率。更多还原

【Abstract】 Objective To investigate the role of miR-34 a in regulating cancer stem cell(CSC) phenotype of nasopharyngeal squamous cell carcinoma in vitro. Methods The cells were isolated from patients with nasopharyngeal carcinoma(NPC) and cultured in vitro. Flow-cytometry was used to quantify and enrich for ALDH~+ cancer stem cells(CSCs). RTqPCR was performed to analyze expression patterns of miR-34 a and CSC-related transcription factors(CSC-TFs)(Sox2,Nanog, Oct3/4) for ALDH~+ and ALDH~- cells. Transfection of miR-34 a mimics was used to evaluate its regulatory potential for CSC marker profiles as well as CSC-TFs expression in NPC-CSC. Results The expression of ALDH was found in around 15.43% of primary NPC cells. miR-34 a expression levels were significantly downregulated in the majority of ALDH~+ cells derived from primary NPC as compared to ALDH-cells(-13.2 vs-4.1 fold). For CSC-related TF expression, ALDH~+ cells showed a significantly increased level compared to ALDH~- cells(up to 36.8 fold). miR-34 a mRNA level in ALDH~+ cells after transfection with miR-34 a mimics significantly increased after 24 h, 48 h and 72 h, and the peak of miR-34 a level was found 48 h post-transfection(4.49 fold, P<0.01). Transfection of miR-34 a mimics significantly reduced the CSC-related TF expression level in ALDH~+ cells. Conclusion Restoration miR-34 a significantly inhibited the formation of CSC-phenotype in human primary NPC cells. Therapeutic modulation of miR-34 a in NPC and CSCs may reduce the rate of metastasis and recurrence of tumors after therapy.更多还原