SIRT4对前列腺癌细胞增殖和侵袭的影响及其机制研究
发布时间:2021-08-17 21:31
背景:前列腺癌是全球最常被诊断的癌症之一,并且是美国男性因癌症死亡的第二大病因。与白人男子相比,非洲男子更容易出现前列腺癌的早期病变,且较容易被诊断出来,而且死亡率更高,且随年龄增长前列腺癌的发病率增加。2018年全球登记的新发前列腺癌约占男性所有新发癌症病例的7.1%。尽管前列腺癌经过早期诊断和有效治疗(如手术和化疗)可提高患者生存率,但前列腺癌仍然是全球面临的主要健康问题。因此,探索前列腺癌致病的分子机制对于开发新颖、有效的前列腺癌治疗非常重要。Sirtuin家族由SIRT1-SIRT7组成,在细胞凋亡、线粒体生物合成、脂质代谢、脂肪酸氧化、细胞应激反应、胰岛素分泌和衰老等不同的细胞过程中发挥重要作用。有研究表明SIRT4可通过抑制谷氨酰胺过表达来抑制和抑制诸如结肠直肠和胃肿瘤等肿瘤细胞的生长、增殖和转化,但目前尚无研究对其功能在前列腺癌发生发展中的作用。目的:明确SIRT4在前列腺癌细胞系中的表达情况,并研究其在前列腺癌细胞中增殖和侵袭的作用。方法:应用实时定量逆转录聚合酶链反应(RT-qPCR)技术检测正常细胞与LNCaP细胞中SIRT4基因表达情况。通过细胞转染技术将SIRT...
【文章来源】:江苏大学江苏省
【文章页数】:56 页
【学位级别】:硕士
【文章目录】:
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
本文编号:3348537
【文章来源】:江苏大学江苏省
【文章页数】:56 页
【学位级别】:硕士
【文章目录】:
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
本文编号:3348537
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