PRL促进乳腺癌细胞增殖及与microRNA表达谱变化的相关性研究  

Correlational study between prolactin promoting breast cancer cell proliferation and microRNA expression profiling change

作　　者：何伟[1] 魏钦俊[1] 刘然[1] 姚俊[1] 鲁雅洁[1] 曹新[1] 

机构地区：[1]南京医科大学生物技术系,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2013年第3期303-307,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81172142);江苏高校优势学科建设工程资助

摘　　要：目的:探讨基于催乳素(prolactin,PRL)反应的人乳腺癌细胞microRNA(miRNA)表达谱变化,以了解其在乳腺癌发生和发展中的潜在作用。方法:人重组PRL处理高表达催乳素受体(PRLR)的人乳腺癌T-47D细胞系,MTT法检测细胞增殖能力变化,流式细胞术检测细胞周期和凋亡变化情况。利用Solexa高通量测序技术检测PRL处理组和未处理组的T-47D细胞miRNAs表达谱,荧光定量PCR法进行初步验证,同时进行相关的生物信息学分析。结果:PRL作用于T-47D细胞后,MTT结果显示细胞增殖能力增强,细胞周期检测结果表明PRL处理后,G1期的细胞比例减少,S期的细胞比例升高,而G2/M期没有变化。细胞凋亡分析结果则显示PRL可抑制细胞凋亡,导致细胞凋亡率下降。通过Solexa高通量测序技术成功检测了PRL处理和未处理组的人乳腺癌T-47D细胞miRNA表达谱,两组中分别检测到821个和798个miRNAs成熟体,其中428个共表达,42个为两组间明显差异表达。选取的4个miRNAs分子经荧光定量PCR法证实其细胞内的表达与测序结果一致。两组miRNA表达谱中同时也检测到86个和115个novel miRNAs成熟体,其中46个novel miRNAs在两组细胞中共表达。结论:在初期实验明确PRL对人乳腺癌T-47D细胞产生促增殖作用的基础上,利用Solexa高通量测序技术检测两类T-47D细胞miRNAs表达谱,获得了一系列与PRLR信号通路高度相关的miRNAs分子,为进一步研究miRNAs分子在人乳腺癌发生发展中的作用奠定重要的研究基础。Objective ：To explore changes of microRNA expression profiling in human breast cancer cell based on prolactin（PRL） treatment,and to understand their potential roles in breast cancer initiation and progression. Methods：Human breast cancer T-47D cells, in which PRL receptor （PRLR） is highly expressed, were treated with or without human recombinant PRL, two group cells were subjected to detect their proliferation, cell cycle and apoptosis alteration by the MTT assay and flow cytometry method （FCM）. miRNA expression profile of PRL treated and untreated cells were analyzed by Solexa sequencing technology, miRNAs were validated by real- time PCR and undertaken bioinformatic analysis. Results：In MTF results,it showed that T-47D cell proliferation ability was enhanced by PRL treatment. FCM revealed that G1 phase cells were reduced,while S phase cells were increased. Meanwhile, cell apoptotic rate was reduced in PRL group. Two miRNAs expression profiles were successfully analyzed from PRL-treated and untreated T-47D cells by Solexa sequencing technology. Within the Solexa sequencing resuhs,821 and 798 miRNAs were detected in the two libraries, respectively. 428 miRNAs were co-expressed, and 42 miRNAs were significantly differentially expressed. Four miRNAs were validated by real-time PCR,their intracellular expression was consistent with Solexa sequencing data. Besides, 86 and 115 novel miRNAs were also detected in the two libraries,and 46 novel miRNAs were co-expressed. Conclusion：PRL can enhance breast cancer cell proliferation ability,and miRNAs expression profiles in T-47D cells treated with or without PRL are successfully analyzed by Solexa sequencing technology. Series of PRLR signaling pathway related miRNAs were harvested. This study provides a reference forelucidating the complex miRNA-mediated regulatory networks of PRL/PRLR signaling pathway that affect breast cancer tumorigenesis and progression.

关 键 词：乳腺癌 Solexa高通量测序 催乳素 MICRORNA 

分 类 号：R737.9[医药卫生—肿瘤]