肿瘤转移相关基因MTA1与鼻咽癌恶性程度和转移的相关分析

发布时间：2018-03-24 15:39

本文选题：血管内皮生长因子　切入点：肿瘤转移相关基因MTA1　出处：《武汉大学》2012年博士论文

【摘要】：第一部分人鼻咽癌中肿瘤转移相关基因MTA1与VEGF、PCNA表达的相关性研究目的：研究人鼻咽癌中肿瘤转移相关基因MTA1与血管内皮生长因子(VEGF)、增殖细胞核抗原(PCNA)表达的相关关系,并结合文献探讨其意义。方法：采用免疫组化(SABC)和图像分析技术检测10例正常鼻咽部组织和75例人鼻咽癌标本中肿瘤转移相关基因MTA1与VEGF、PCNA表达水平,并分析表达的相关性。结果：(1)肿瘤转移相关基因MTA1、VEGF、PCNA在正常鼻咽部组织中的表达均为阴性；(2)肿瘤转移相关基因MTA1、VEGF、PCNA在人鼻咽癌中均有阳性表达,(3)人鼻咽癌中肿瘤转移相关基因MTA1与VEGF、PCNA的表达水平呈明显正相关(r=0.941和r=0.889,P0.05)。结论：肿瘤转移相关基因MTA1在人鼻咽癌的恶性生物学行为中可能起重要的作用,其可能成为人鼻咽癌治疗的一个有效的靶点。第二部分肿瘤转移相关基因MTA1与鼻咽癌细胞浸润和转移的关系目的：鼻咽癌具有很高的侵润和转移特性。而肿瘤转移相关基因(MTA1)是新近发现的一个肿瘤转移候选基因。本研究通过比较MTA1基因在人鼻咽癌细胞高低转移株的表达水平,探讨MTA1表达与鼻咽癌细胞浸润和转移潜能的相关性。方法：采用半定量逆转录聚合酶链反应(RT-PCR)检测增殖及转移潜能不同的鼻咽癌细胞株6-10B和5-8F中MTA1的表达情况,用Boyden小室体外侵袭实验检测两株细胞的体外转移能力；用MTA1基因质粒转染低转移潜能细胞株6-10B,通过半定量逆转录聚合酶链反应检测MTA1的表达；并检测转染前后细胞转移能力的变化。结果：MTA1在低转移潜能细胞株6-10B中表达水平低,在高转移潜能细胞株5-8F中表达水平高(p0.05)；体外侵袭实验显示高转移株5-8F细胞体外侵袭力强(穿膜细胞相对百分率为46.3+2.4%)显著高于低转移潜能细胞株,其穿膜细胞相对百分率只有12.6+1.1%,转染MTA1基因质粒后,低转移细胞株转移潜能较未转染细胞明显增高(p0.05)。结论：MTA1在人鼻咽癌细胞转移过程中起重要作用,其作用机制以及作为肿瘤转移治疗靶点的可能性值得进一步研究。第三部分反义寡核苷酸对鼻咽癌细胞MTAl基因表达及侵袭性影响的研究目的：合成人肿瘤转移相关基因(MTA1)的反义脱氧寡核苷酸,观察其转染后对人鼻咽癌CNE1细胞系中MTA1表达及鼻咽癌侵袭性的影响。方法：免疫细胞染色观察人工合成正义、反义及无意义MTA1基因片段转染人鼻咽癌细胞CNE1后人肿瘤转移相关基因的表达,RT-PCR和Western blot检测MTA1基因的(?)nRNA和蛋白质的表达水平的变化。应用Boyden小室体外侵袭力评价转染前后细胞侵袭力的变化。结果：反义寡核苷酸处理鼻咽癌细胞后,MTA1mRNA的表达下降(吸光度之比为24.09±0.22),与正义链组、无意义链组和空白对照组(34.23+0.15,33.22±0.28,35.18±0.22)相比差异有统计学意义,P0.05。Boyden小室体外侵袭实验检测显示,反义寡核苷酸处理后鼻咽癌细胞的透膜能力明显下降。结论：MTA1同鼻咽癌的转移能力密切相关,反义寡核苷酸的转染可阻遏鼻咽癌细胞中人肿瘤转移相关基因(MTA1)的表达。并因此有可能限制鼻咽癌的侵袭和转移。
[Abstract]:The correlation of tumor metastasis related gene MTA1 and VEGF, PCNA expression in human nasopharyngeal carcinoma
Objective: To investigate the correlation between tumor metastasis related gene MTA1 and vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) expression in human nasopharyngeal carcinoma, and to explore its significance combined with literature.
Methods: immunohistochemistry and SABC were used to detect the expression levels of MTA1 and VEGF and PCNA in 10 normal nasopharyngeal tissues and 75 NPC cases, and the correlation between them was analyzed.
Results: (1) tumor metastasis related genes MTA1, VEGF, PCNA expression in normal nasopharyngeal tissues were negative; (2) tumor metastasis related genes MTA1, VEGF, PCNA were positive in human nasopharyngeal carcinoma, (3) human nasopharyngeal carcinoma metastasis related genes MTA1 and VEGF showed a positive correlation the expression level of PCNA (r=0.941, r=0.889, P0.05).
Conclusion: tumor metastasis related gene MTA1 may play an important role in malignant biological behavior of human nasopharyngeal carcinoma. It may become an effective target for treatment of NPC.
The relationship between the second part of tumor metastasis related gene MTA1 and the invasion and metastasis of nasopharyngeal carcinoma cells
Objective: nasopharyngeal carcinoma with invasion and metastasis is very high. And the characteristics of tumor metastasis associated gene (MTA1) is a recently discovered tumor metastasis genes. The expression level of MTA1 gene in human nasopharyngeal carcinoma cell lines with different metastatic cells, to investigate the correlation between MTA1 expression and the potential of invasion and metastasis of nasopharyngeal carcinoma.
Methods: using semi quantitative reverse transcriptase polymerase chain reaction (RT-PCR) to detect the proliferation and metastatic potential of different nasopharyngeal carcinoma cell lines 6-10B and MTA1 in 5-8F expression, with Boyden chamber in vitro invasion assay of two cell lines in vitro metastasis; MTA1 gene transfected with low metastatic potential cell line 6-10B was detected by semi quantitative MTA1. Reverse transcriptase polymerase chain reaction; change detection and transfer ability before and after transfection.
Results: the MTA1 in low metastatic potential cell line 6-10B low expression in highly metastatic potential cell line 5-8F high expression level (P0.05); experiments show high metastatic 5-8F cells in vitro invasiveness in vitro invasion (transmembrane cell relative percentage of 46.3+2.4%) was significantly higher than that of low metastatic potential cell line, the cell membrane the relative percentage of only 12.6+1.1%, transfection of MTA1 gene plasmid, metastatic potential than non transfected cells was significantly higher in low metastatic cell line (P0.05).
Conclusion: MTA1 plays an important role in the metastasis of human nasopharyngeal carcinoma cells, and its mechanism and the potential as a therapeutic target for tumor metastasis are worth further research.
Study on the effect of third antisense oligonucleotides on MTAl gene expression and invasiveness in nasopharyngeal carcinoma cells
Objective: to synthesize antisense oligodeoxynucleotide of human tumor metastasis related gene (MTA1) and observe its effect on MTA1 expression and invasion of nasopharyngeal carcinoma cell line after transfection in human nasopharyngeal carcinoma CNE1 cell line.
Methods: To observe the effect of synthetic justice immune cells staining. The expression of antisense MTA1 gene transfection and significance of human nasopharyngeal carcinoma cells CNE1 human tumor metastasis related gene, detection of MTA1 gene RT-PCR and blot Western (?) expression of nRNA and protein. The application of Boyden chamber invasion force evaluation changes before and after transfection cell invasion.
Results: antisense oligonucleotide treatment of nasopharyngeal carcinoma cells, the expression of MTA1mRNA decreased (absorbance ratio of 24.09 + 0.22), and sense group, no significant chain group and blank control group (34.23+0.15,33.22 + 0.28,35.18 + 0.22) compared to the difference was statistically significant, P0.05.Boyden chamber in vitro invasion assay showed that antisense oligonucleotide treatment membrane ability of nasopharyngeal carcinoma cells decreased significantly.
Conclusion: MTA1 is closely related to the metastatic ability of nasopharyngeal carcinoma. The transfection of antisense oligonucleotides can inhibit the expression of human tumor metastasis related gene (MTA1) in nasopharyngeal carcinoma cells. Therefore, it may limit the invasion and metastasis of nasopharyngeal carcinoma.

【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R739.63

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