SFRP5沉默对人胆管癌细胞生物学特性的影响

兰州大学机构库 >第一临床医学院

	SFRP5沉默对人胆管癌细胞生物学特性的影响
Alternative Title	Effects of silencing SFRP5 to the cellular property in human cholangiocarcinoma RBE cells
	李力宏
Subtype	硕士
Thesis Advisor	周文策
	2016-05-19
Degree Grantor	兰州大学
Place of Conferral	兰州
Degree Name	硕士
Keyword	SFRP5 胆管癌信号传导靶基因修复
Abstract	探讨使用慢病毒介导短发夹RNA沉默SFRP5基因对人类胆管癌RBE细胞株增殖、迁移以及细胞周期和细胞凋亡的影响。对人胆管癌RBE细胞株进行体外培养，制备设计SFRP5干扰片段，使用慢病毒载体pLVX-shRNA2构建慢病毒载体质粒，并进行基因测序鉴定，将构建好的慢病毒shRNA载体以及辅助质粒共同转染293T细胞获得重组慢病毒, 利用梯度测定法测定慢病毒滴度，使用FuGENE® HD转染试剂将慢病毒转染入RBE细胞株，获得稳定转染的RBE细胞株，通过Transwell细胞迁移实验以及细胞划痕实验研究SFRP5沉默对胆管癌细胞的迁移能力的变化，使用CCK-8法检测SFRP5低表达对胆管癌细胞增殖水平的影响，利用流式细胞术检测SFRP5低表达对胆管癌细胞周期以及细胞凋亡的影响。实验结果发现SFRP5可能在肝内胆管癌的发生发展中起到重要的调节作用，SFRP5低表达激活wnt信号通路，促进肿瘤的发生发展。人肝内胆管癌RBE细胞株中，沉默SFRP5基因表达可显著增强RBE细胞的增殖和迁移能力，SFRP5的沉默抑制细胞凋亡，SFRP5在胆管癌细胞RBE中起到抑癌作用。
Other Abstract	To investigate the silence efficiency of lentivirus-mediated shRNA(short hairpin RNA)silencing SFRP5 on proliferation, migration and apoptosis of cholangiocarcinoma RBE cells. In our research,The human cholangiocarcinoma RBE cell line were cultured in vitro,In the present work, We designed the interference sequence to amplify the coding region of SFRP5, The plasmid was verification by DNAsequencing, after transfected the sFRP-5 knockdown vector(pLVX-shRNA2) into RBE cells by using the FuGENE® HD Transfection Reagentas the cell conversion agent, we obtained a RBE cell line with stable SFRP5 knockdown, which was validated by reverse transcription quantitative polymerase chain reaction and western blotting. The CCK-8 assay was used to measure the capability of cells proliferation, The wound healing assay and transwell migration assay was used to exploring the effects of SFRP5 knockdown on cell motility, the cell cycle and apoptosis was measured by using flow cytometry.SFRP5 may play an important regulatory role in the development of intrahepatic cholangiocarcinoma, low expression of SFRP5 activate wnt signaling pathway, and promote tumor development. After sucessfully constructed a RBE Cell Line with stable knockdown of SFRP5, our results indicate that SFRP5 knockdown enhances proliferation and migration potential of RBE cells in vitro, and can decrease cell apoptosis at the same time. Our results suggest SFRP5 might act as tumor suppressor in cholangiocarcinoma RBE cells, and the present study lay the foundation for the future experiments in vitro.
URL	查看原文
Language	中文
Document Type	学位论文
Identifier	https://ir.lzu.edu.cn/handle/262010/201218
Collection	第一临床医学院
Recommended Citation GB/T 7714	李力宏. SFRP5沉默对人胆管癌细胞生物学特性的影响[D]. 兰州. 兰州大学,2016.

Files in This Item:
There are no files associated with this item.

If you have any objections to this item, please fill out the form below and the administrator will contact you as soon as possible.
Content:
Email：	*
Institution:
Verification Code:	Refresh

Any comments and suggestions are welcomed.
Title:	*
Content:
Email：	*
Verification Code:	Refresh

第一临床医学院知识库