TIP30诱导肝癌细胞凋亡信号转导途径的研究

摘要

该研究中,我们用带有人类野生型TIP30基因的复制缺陷型腺病毒载体感染人肝癌细胞系;HepG2(p53基因野生型)、Hep3B(p53基因缺陷型)和PLC/RPF/5(p53基因突变型),探讨TIP30凋亡调控的分子机制.该研究从以下几个方面入手:1.TIP30引起的细胞凋亡形态变化.2.TIP30对一些凋亡相关基因的调节,如对p53,Bax,Bad,Bcl-xl的调节.3.TIP30是否介导线粒体凋亡信号转导途径:包括线粒体膜位变化、对下游凋亡调节因子caspase-9,-3和PARP的激活.阐明TIP30参与凋亡的分子机制将为其在基因治疗的临床应用奠定理论基础.结论:1.通过对TIP30介导的肝癌细胞的凋亡率的检测,我们确定:TIP30可引起肝癌细胞的凋亡.TIP30为一公认的肿瘤生长抑制因子.2.在HepG2细胞中,TIP30介导的凋亡信号转导途径为:上凋p53后促进Bax和Bad从细胞质向线粒体的跃迁,引起细胞色素c的释放.然后激活caspase9,3和剪切PARP.因而TIP30介导的凋亡信号转导途径为p53/mitochondria/caspase-9途径.3.在无p53功能的Hep3B和PLC/RPF/5中,TIP30仍然能诱导细胞凋亡.说明TIP30诱导的p53依赖型凋亡只存在于某些细胞中,这些细胞中可能存在与TIP30起作用的其他因子,TIP30可能通过这些因子促进Bad的跃迁、下调Bcl-xl和剪切PARP.虽然线粒体凋亡途径已被深入研究,但其上游调节因子还有待于发现,该研究为肝癌的治疗提供了可靠的理论依据.

目录

原创性声明及关于学位论文使用授权的声明

Abstract in English

ABBREVIATLON

Review The mitochondrial pathway during cell death signaling

Apoptosis outline

Mitochondria

Release of Cyt c and Other Mitochondrial Proteins during Apoptosis

Factors affecting the release of cytochrome C

1.Role of the Bcl-2 Family

2.Engagement of the mitochondria through p53 and other factors

Changes of mitochondria

Caspaes

Conclusion

Reference

Section 1 The construction, appraisal and extraction of Ad-TIP30

Introduction

Materials and equipments

Methods

Results

Discussion

References

Section2 TIP30 regulates the expression of apoptosis-related genes in human hepatocellular carcinoma cells undergoingp53/ mitochondria apoptotic signal introduction pathway

Introduction

Materials and equipments

Results

Disscusion

References

Section 3 Regulation of Bcl-2 family in TIP30-induced apoptosisin Hepatoblastoma cells

Introduction

Materials and equipments

Methods

Results

Discussion

References

Section 4 Apoptosis from activation of p53 and regulation of members of Bcl-2 family to mitochondrial release of cytochrome cin Hepatoblastoma cells

Introduction

Materials and equipments

Methods

Results

Discussion

References

Conclusion

Articles published

Acknowledgements

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