nm23-H1与MMP-2、MMP-9在子宫内膜异位症中的表达及相关性的研究

发布时间：2018-11-20 17:21

【摘要】：目的:子宫内膜异位症是育龄期妇女中常见的慢性疾病,其发病机制尚不明确。子宫内膜异位症的临床表现主要为继发性痛经、慢性盆腔痛、不孕等,严重影响着人们的生活。子宫内膜异位症本身虽是一种良性疾病,却具有恶性肿瘤粘附、局部浸润和远处转移的特点。MMPs作为肿瘤转移相关基因,参与多种肿瘤的发病过程,我们推测MMPs也参与了子宫内膜异位症的发生发展过程。现已有大量研究提示nm23-H1可通过RAS途径抑制MMPs的表达,抑制细胞的转移能力,从而参与多种肿瘤的发生、发展过程。前期实验已证实nm23-H1在子宫内膜异位症的异位内膜组织中的表达明显低于正常子宫内膜组,推测nm23-H1在子宫内膜异位症的发病过程中起到一定作用,然而在子宫内膜异位症中nm23-H1是否可通过影响MMPs的表达变化来影响子宫内膜组织的侵袭、转移能力尚未得到证实。本实验旨在通过分析MMP-2、MMP-9在正常子宫内膜组织及异位子宫内膜组织中的表达变化,探讨MMP-2、MMP-9在子宫内膜异位症发生、发展中的可能起到的作用,并结合前期试验结果分析nm23-H1与MMP-2、MMP-9在子宫内膜异位症发生、发展中是否存在相关性,探讨nm3-H1是否可通过RAS信号通路影响子宫内膜异位症的发展。方法:将收集到的正常子宫内膜组织38例及异位子宫内膜组织45例均分为两份,其中1份用Trizol一步法提取组织总RNA,并将其逆转录成cDNA,采用实时荧光定量PCR技术定量检测其中的MMP-2、MMP-9基因在正常和异位子宫内膜组织中m RNA的表达情况;另一份提取组织总蛋白,采用Western Blot方法,检测MMP-2、MMP-9基因在正常和异常子宫内膜组织中蛋白的表达情况。结果:1、qRT-PCR实验结果:(1)子宫内膜异位症组织中MMP-2基因的mRNA表达量为4.01±2.89,在正常子宫内膜组MMP-2基因mRNA的表达量为1.40±1.13,将两组中MMP-2基因mRNA的表达比较可发现,异位内膜组织中的表达明显高于正常子宫内膜组,,差异有统计学意义(t=-3.66,P=0.000,P0.05)。(2)子宫内膜异位症组织中MMP-9基因的mRNA表达量为3.04±2.77,正常子宫内膜组MMP-9基因mRNA的表达量为0.99±0.86,将两组中MMP-9基因mRNA的表达比较可发现,异位内膜组织中的表达明显高于正常子宫内膜组,差异有统计学意义(t=-3.158,P=0.000,P0.05)。(3)结合前期试验,对nm23-H1与MMP-2、nm23-H1与MMP-9相关性的分析,发现前者r 1=-0.21,P0.05;后者r 2=-0.12,P0.05,提示在子宫内膜异位症中nm23-H1与MMP2、MMP-9存在负性相关,但两两之间相关性不强。2、Western Blot实验结果:经研究发现MMP-2在子宫内膜异位症的异位内膜组织中的蛋白表达为水平为1.64±0.62,正常子宫内膜组MMP-2的蛋白表达量为0.61±0.31,两者比较可见,MMP-2在异位子宫内膜组织中的表达明显高于正常子宫内膜组,两者间的差异有统计学意义(t=-0.75,P=0.027,P0.05)。结论:MMP-2、MMP-9基因在异位子宫内膜组织中表达上调,提示MMP-2、MMP-9在子宫内膜异位症的发生、发展中起到一定的作用。结合前期实验分析发现在子宫内膜异位症中nm23-H1基因与MMP-2、MMP-9基因表达呈负相关,但他们之间的相关性不强,故而若为验证nm23-H1基因是否通过RAS途径抑制MMP-2、MMP-9在异位内膜组织中的表达需进行进一步研究。有关nm23-H1与MMP-2、MMP-9基因的在子宫内膜异位症的相关性研究,有助于促进对内异症的进一步认识,为子宫内膜异位症的诊断、治疗及预后提供新的标靶。
[Abstract]:Objective: Endometriosis is a common chronic disease in women of childbearing age, and its pathogenesis is not clear. The clinical manifestations of endometriosis are secondary dysmenorrhea, chronic pelvic pain, and infertility. Endometriosis itself is a benign disease, but has the characteristics of malignant tumor adhesion, local infiltration and distant metastasis. MMPs, as a tumor metastasis-related gene, are involved in the pathogenesis of multiple tumors, and we assume that MMPs also participate in the development of endometriosis. It has been shown that nm23-H1 can inhibit the expression of MMPs by RAS, and inhibit the transfer ability of cells, thus participating in the occurrence and development of multiple tumors. The expression of nm23-H1 in the ectopic endometrial tissue of endometriosis is significantly lower than that of the normal endometrium, and it is presumed that the nm23-H1 plays a role in the pathogenesis of endometriosis. However, whether nm23-H1 in endometriosis can influence the invasion and metastasis of the endometrial tissue by influencing the expression change of MMPs has not been confirmed. The purpose of this study was to study the expression of MMP-2 and MMP-9 in the normal endometrium and the tissue of the ectopic endometrium, and to explore the possible role of MMP-2 and MMP-9 in the occurrence and development of endometriosis, and to analyze the expression of nm23-H1 and MMP-2 in the early-stage test. MMP-9 is related to the occurrence and development of endometriosis, and whether nm3-H1 can influence the development of endometriosis through the RAS signal pathway. Methods: A total of 38 cases of normal endometrium and 45 cases of ectopic endometrium were divided into two groups. One of them was used to extract total RNA from the tissue by Trizol one-step method and reverse transcription into cDNA. The expression of MMP-2 was detected by real-time fluorescence quantitative PCR. The expression of MMP-9 gene in normal and ectopic endometrium was detected by Western Blot method, and the expression of MMP-2 and MMP-9 in normal and abnormal endometrial tissues was detected. Results: (1) The mRNA expression of MMP-2 gene in the tissue of endometriosis was 4.01-2.89, and the expression of MMP-2 mRNA in the normal endometrium was 1. 40-1.13, and the expression of MMP-2 mRNA in the two groups could be found. The expression of ectopic endometrium was significantly higher than that of normal endometrium (t =-3.66, P = 0.000, P0.05). (2) The expression of MMP-9 in the tissue of endometriosis was 3.04-2.77, and the expression of MMP-9 mRNA in the normal endometrium was 0.99-0.86. The expression of MMP-9 mRNA in the two groups was found to be higher than that of the normal endometrium. The difference was significant (t =-3.158, P = 0.000, P0.05). (3) The correlation between nm23-H1 and MMP-2, nm23-H1 and MMP-9 was analyzed in the early stage. The results showed that there was negative correlation between nm23-H1 and MMP2 and MMP-9 in endometriosis. The results showed that the expression of MMP-2 in the ectopic endometrium of endometriosis was 1.64-0.62, and the expression of MMP-2 in the normal endometrium was 0.61-0.31. The expression of MMP-2 in the tissue of the ectopic endometrium was significantly higher than that of the normal endometrium. The difference between the two groups was statistically significant (t =-0.75, P = 0.027, P0.05). Conclusion: The expression of MMP-2 and MMP-9 in ectopic endometrium is up-regulated, suggesting that MMP-2 and MMP-9 play a role in the pathogenesis and development of endometriosis. The expression of nm23-H1 gene in endometriosis was negatively correlated with the expression of MMP-2 and MMP-9, but the correlation between them was not strong, and the expression of MMP-2 and MMP-9 in the tissue of ectopic endometrium was further studied. The study of the correlation between nm23-H1 and MMP-2 and MMP-9 in endometriosis can help to promote the further understanding of endometriosis and provide a new target for the diagnosis, treatment and prognosis of endometriosis.
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R711.71

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