通过调控靶基因影响乳腺上皮细胞的增殖.docxVIP

CONTENTSCONTENTS CONTENTS CONTENTS Abstract in Chinese ·I Abstract in English ．．III 1 Introduction 1 1．1 The research progress ofmiRNAs ·1 j．1，1 The discovery and naming progress ofmiRNAs 1 1．1．2 The biosynthesisprogress ofmiRNAs ·1 1．1．3 The biological characteristics and functions ofmiRNAs 2 1．1．4 The detection Technology ofmiRNAs 一2 1．2 The target gene search and verification ofmiRNAs ··3 1．2．1 The target gene bioinformatics prediction ofmiRNAs ··3 1．2．2 The target gene validation ·-3 1．3 The research progress ofmiR-142-3p ·5 1．4 The research progress ofgene尸冲Ⅳ，J ·‘5 1．5 The regulation ofmiRNA tumor ·7 1．6 The purpose and significance ofthe study ． ”8 2 MateriaIs and methods ．9 2．1 Major experimental materials ·9 2．2 Major instrumen 9 2．3 Major agents 10 2．4 Preparation for major agents ·10 2．5 Experimental method and step ． 1 1 2．5．1 Screening the target genes ofmiR-142_3p 1 1 2．5．2 The culture ofmammary epithelial cells 12 2．5．3 The influences mammary epithelial cells MCF-10A after overexpression of miR．142．3p 12 2．5．4 The influences mammary epithelial cells after inhibition ofmiR-142·3p 17 2．5．5 The influences of the overpress of miR．142—3p mammary epithelial cells MCF-10A aRer the silencing ofPTPNll 18 2．5．6 Statistical analysis 19 3 Results ．20 3．1 Screening ofthe target genes 20 3．1．1 Analysis ofthe results ofRNA．protein immunopreciptation ·20 3．1．2 The action sites ofPTPNll and miR-142—3p 22 3．2 The influences mammary epithlial cells MCF一10A after overexpression ofmiR-142-3p·22 万方数据 东北农业大学农学硕士学位论文3．2．1 东北农业大学农学硕士学位论文 3．2．1 Detect the expression ofmiR．142．3p via RNA Nano Detection Technology in live cell after overexpression ofmiR-142-3p ··22 3．2。2 The influences the expression of gene PTPN n in mammary epithelial cells after inducing the mimics of miR-142—3p ··23 3．2．3 The influences gene PTPNl 1 and other proteins relative to proliferation in mammary epithelial cells after inducing the mimics of miR一142·3p ．23 3．2．4 Immunohistochemistry ”25 3．2．5 The i