CPT1A促进食管癌细胞侵袭运动的分子机制研究

发布时间：2018-03-22 22:33

本文选题：食管鳞癌　切入点：CPT1A　出处：《昆明理工大学》2017年硕士论文　论文类型：学位论文

【摘要】：食管癌进展快、预后差,其死亡率居我国恶性肿瘤第四位。尽管手术及放化疗技术取得了巨大的进步,但是食管癌患者的5年生存率仍然较低。因此鉴定食管癌变过程中的关键调控分子并揭示其调控机制,对于改善食管癌的诊断和治疗具有重要意义。我们前期研究发现,CPT1A的拷贝数增加与食管癌患者不良预后正相关。免疫组化的结果表明CPT1A高表达与食管癌患者的不良预后正相关,且是独立预后判断因素。体外实验发现,敲降CPT1A可以显著抑制KYSE450和KYSE510细胞的侵袭和转移能力。为此我们根据基因表达谱芯片结果,进一步鉴定调控食管癌细胞转移和侵袭的关键基因。在本研究中,我们通过RT-PCR和蛋白免疫沉淀法发现敲降CPT1A可以下调uPA的mRNA和蛋白表达水平。深入研究发现,敲降CPT1A可以下调NF-κB p65的磷酸化水平,并且发现敲降NF-κB p65显著抑制uPA的转录和肿瘤细胞的侵袭和迁移能力。PKCiota和P62是NF-κB信号通路的上游调控基因。我们研究进一步发现敲降CPT1A可以下调PKCiota和P62的表达水平,并且PKCiota与p62之间存在反馈调节作用。在KYSE450细胞中,敲降PKCiota可以下调P62和uPA的表达水平,以及NF-κB p65的磷酸化水平。而加入泛素-蛋白酶体途径抑制剂MG132可以显著回复敲降PKCiota和CPT1A所引起的P62表达水平下调。有趣的是,我们发现敲降NF-κB p65也可以下调CPT1A的表达水平。综上所述,我们得出结论CPT1A通过调控PKCiota/p62/NF-KB/uPA信号轴促进食管癌细胞的侵袭和转移能力。本论文进一步阐明了 CPT1A促进食管细胞转移和侵袭的分子机制,并有望成为食管鳞癌治疗的新的分子靶点。
[Abstract]:Esophageal cancer is developing rapidly and its prognosis is poor. Its mortality rate is the fourth highest in China. Despite the great progress made in surgery and radiotherapy and chemotherapy, But the 5-year survival rate of esophageal cancer patients is still low. It is important to improve the diagnosis and treatment of esophageal cancer. Our previous study found that the increased copy number of CPT1A was positively related to the poor prognosis of esophageal cancer patients. The immunohistochemical results showed that the high expression of CPT1A was positively correlated with the poor prognosis of esophageal cancer patients. In vitro, knock down CPT1A could significantly inhibit the invasion and metastasis of KYSE450 and KYSE510 cells. In this study, we found that knockout CPT1A could down-regulate the expression of mRNA and protein in uPA by RT-PCR and protein immunoprecipitation. Knockdown of CPT1A can down-regulate phosphorylation level of NF- 魏 B p65. It was also found that knockout NF- 魏 B p65 significantly inhibited uPA transcription and invasion and migration of tumor cells. PKCiota and P62 were upstream regulatory genes of NF- 魏 B signaling pathway. We further found that knockdown CPT1A could down-regulate the expression of PKCiota and P62. There is feedback regulation between PKCiota and p62. Knockout PKCiota can down-regulate the expression of P62 and uPA in KYSE450 cells. And the phosphorylation level of NF- 魏 B p65. The down-regulation of P62 expression induced by knockout PKCiota and CPT1A was significantly reversed by the addition of ubiquitin proteasome pathway inhibitor MG132. We found that knocking down NF- 魏 B p65 could also down-regulate the expression of CPT1A. We conclude that CPT1A can promote the invasion and metastasis of esophageal cancer cells by regulating PKCiota/p62/NF-KB/uPA signal axis. In this paper, we further elucidate the molecular mechanism of CPT1A promoting esophageal cell metastasis and invasion. It is expected to be a new molecular target for the treatment of esophageal squamous cell carcinoma.
【学位授予单位】：昆明理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.1

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