糖尿病与非糖尿病小鼠子宫内膜蛋白质组学及LDHB的表达和功能的研究

发布时间：2018-05-24 18:34

本文选题：子宫内膜癌 + 2型糖尿病　；参考：《华中科技大学》2014年博士论文

【摘要】：目的：根据糖尿病组(db/db小鼠)和正常对照组(db/m小鼠)子宫内膜蛋白质差异表达谱,寻找可能的联系肥胖、2型糖尿病及子宫内膜癌的纽带分子。进一步,对筛选出的差异蛋白LDHB,了解其在人子宫内膜癌组织中的表达及临床意义,再进行细胞水平的功能研究。方法：采用二维蛋白电泳及蛋白质谱检测糖尿病组及非糖尿病组小鼠子宫内膜的蛋白质差异表达普,并应用western及免疫组织化学方法对选择的三个蛋白进行了鉴定。应用HE染色方法,比较糖尿病小鼠与非糖尿病小鼠子宫内膜的厚度情况。用免疫组织化学的方法了解LDHB在子宫内膜癌中的表达及临床意义。采用瞬时转染shRNA质粒的方法,干扰ishikawa中LDHB的表达,检测其迁移及侵袭能力的改变。结果：糖尿病小鼠与非糖尿病小鼠子宫内膜差异蛋白质谱中发现了58个差异蛋白,其中上调30个,下调28个,很多蛋白与线粒体功能相关。在此基础上,对其中的三个蛋白：LDHB、 CLIC-1和Calbindin鉴定确定三者都在糖尿病小鼠中的表达高于非糖尿病小鼠。然而,奇怪的是,糖尿病小鼠子宫内膜的厚度较对应月份非糖尿病小鼠的子宫内膜厚度薄很多,可能是不孕的因素之一。进一步,我们发现在子宫内膜癌组织中,LDHB的表达相对缺失,基质中几乎无LDHB的表达,LDHB的表达与肌层浸润深度有关(P=0.003)。在细胞水平,干扰LDHB的表达,增加ishikawa细胞的迁移及侵袭能力(P0.05)。结论：糖尿病小鼠子宫内膜可能伴随线粒体功能异常。LDHB在子宫内膜癌中表达相对缺失,干扰ishikawa细胞中LDHB的表达促进肿瘤细胞的迁移和侵袭,子宫内膜癌的肌层浸润与LDHB的降低有关。
[Abstract]:Aim: according to the differential expression profiles of endometrial proteins between diabetic and normal control group (B / r db mice) and normal control group (n = 10), a possible link between obese type 2 diabetes mellitus and endometrial carcinoma was found. Furthermore, the expression and clinical significance of LDHBs in human endometrial carcinoma were investigated, and the function of LDHBs at cell level was studied. Methods: two dimensional protein electrophoresis and protein spectrum were used to detect the differential expression of proteins in endometrium of diabetic and non-diabetic mice. The three proteins were identified by western and immunohistochemistry. The endometrium thickness of diabetic mice and non-diabetic mice was compared by HE staining. To investigate the expression and clinical significance of LDHB in endometrial carcinoma by immunohistochemical method. The transient transfection of shRNA plasmid was used to interfere with the expression of LDHB in ishikawa and to detect the changes of its migration and invasion ability. Results: 58 differentially expressed proteins were found in endometrium of diabetic and non-diabetic mice, including 30 up-regulated proteins and 28 down-regulated proteins. Many proteins were related to mitochondrial function. On this basis, the expression of three of them in diabetic mice was higher than that in non-diabetic mice. Oddly, however, the thickness of the endometrium in diabetic mice is much thinner than that in non-diabetic mice in the corresponding month, which may be one of the factors contributing to infertility. Furthermore, we found that the expression of LDHB was relatively absent in endometrial carcinoma, and almost no expression of LDHB was related to the depth of myometrial invasion. At the cellular level, it interferes with the expression of LDHB and increases the migration and invasion ability of ishikawa cells. Conclusion: the endometrium of diabetic mice may be accompanied by mitochondrial dysfunction. LDHB expression is relatively absent in endometrial carcinoma, which interferes with the expression of LDHB in ishikawa cells and promotes the migration and invasion of tumor cells. The myometrial invasion of endometrial carcinoma is related to the decrease of LDHB.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R737.33;R587.1

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