背景:前列腺癌是全球最常被診斷的癌癥之一,并且是美國男性因癌癥死亡的第二大病因。與白人男子相比,非洲男子更容易出現(xiàn)前列腺癌的早期病變,且較容易被診斷出來,而且死亡率更高,且隨年齡增長前列腺癌的發(fā)病率增加。2018年全球登記的新發(fā)前列腺癌約占男性所有新發(fā)癌癥病例的7.1%。盡管前列腺癌經(jīng)過早期診斷和有效治療（如手術(shù)和化療）可提高患者生存率,但前列腺癌仍然是全球面臨的主要健康問題。因此,探索前列腺癌致病的分子機(jī)制對于開發(fā)新穎、有效的前列腺癌治療非常重要。Sirtuin家族由SIRT1-SIRT7組成,在細(xì)胞凋亡、線粒體生物合成、脂質(zhì)代謝、脂肪酸氧化、細(xì)胞應(yīng)激反應(yīng)、胰島素分泌和衰老等不同的細(xì)胞過程中發(fā)揮重要作用。有研究表明SIRT4可通過抑制谷氨酰胺過表達(dá)來抑制和抑制諸如結(jié)腸直腸和胃腫瘤等腫瘤細(xì)胞的生長、增殖和轉(zhuǎn)化,但目前尚無研究對其功能在前列腺癌發(fā)生發(fā)展中的作用。目的:明確SIRT4在前列腺癌細(xì)胞系中的表達(dá)情況,并研究其在前列腺癌細(xì)胞中增殖和侵襲的作用。方法:應(yīng)用實(shí)時定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)（RT-qPCR）技術(shù)檢測正常細(xì)胞與LNCaP細(xì)胞中SIRT4基因表達(dá)情況。通過細(xì)胞轉(zhuǎn)染技術(shù)將SIRT... 

【文章來源】：江蘇大學(xué)江蘇省

【文章頁數(shù)】：56 頁

【學(xué)位級別】：碩士

【文章目錄】：
Abstract
摘要
CHAPTER1 INTRODUCTION
    1.1 Background
    1.2 Epidemiology and Incidence
    1.3 Clinical Findings
    1.4 Etiology
    1.5 Grading and Staging
    1.6 Treatment
    1.7 Sirtuin
CHAPTER2 MATERIALS AND METHODS
    2.1 Materials
        2.1.2 Instruments
        2.1.3 Primers
    2.2 Main reagents used and preparation
    2.3 Experimental methods
        2.3.1 Cell culture and growth conditions
        2.3.2 Transfection of human cancer cells with Lipofectamine3000
        2.3.3 Extraction of total cellular proteins
        2.3.5 Total RNA extraction,RNA reverse transcription,and primer verification
            2.3.5.1 Total RNA extraction
            2.3.5.2 RNA reverse transcription
        2.3.6 Quantitative real-time polymerase chain reaction(qRT-PCR)
        2.3.7 SDS-PAGE and Western Blotting
            2.3.7.1 SDS-PAGE
            2.3.7.2 10% separation gel
            2.3.7.3 5% stacking gel
            2.3.7.4 SDS-PAGE electrophoresis
            2.3.7.5 Western Blotting
            2.3.7.6 Immunostaining
        2.3.8 CCK‐8 assay
        2.3.9 Transwell assay
CHAPTER3 RESULT
    3.1 RT-qPCR of SIRT4 gene expression in different prostate cell
    3.2 Results of RT-qPCR after SIRT4 gene over-expression in LNcap cells
    3.3 The growth of LNcap cell transfected with over-expressed SIRT4 gene
    3.4 Effects of SIRT4 on cell proliferation
    3.5 Effect of under-expressed SIRT4 on cell invasion
    3.6 RT-qPCR result of the gene related to SIRT4
    3.7 Western blot result of target genes related to SIRT4
CHAPTER4 DISCUSSION
CHAPTER5 CONCLUSION
REFERENCES
CLINICAL ROTATION
ACKNOWLEDGEMENT
LIST OF PUBLICATIONS
LIST OF ABREVIATIONS



本文編號：3348537