摘要
目的比较正常肾上腺组织、肾上腺皮质腺瘤组织和肾上腺皮质癌组织的蛋白质组学差异,探讨肾上腺皮质肿瘤的鉴别诊断和肾上腺皮质癌的发病、转移机制以及治疗靶点。方法分别提取9例正常肾上腺皮质组织、9例肾上腺皮质腺瘤标本以及9例肾上腺皮质癌标本总蛋白,蛋白经纯化后用固相p H梯度双向凝胶电泳分离肾上腺皮质癌和肾上腺皮质腺瘤组织蛋白质,银染显色,通过PDQUEST 2DE软件分析获取双向凝胶电泳图谱,找出差异表达的蛋白质点。切取差异蛋白质点进行MALDI TOF/TOF质谱鉴定,获取肽质指纹图谱并查询Swiss Prot数据库寻找匹配蛋白质。结果所获得的双向电泳银染图谱重复性较好,正常肾上腺皮质组织蛋白质点约为(2 247±87),肾上腺皮质腺瘤组织蛋白质点数约为(2 375±142),肾上腺皮质癌组织蛋白质点数约为(2 143±107)。在三组组织中,发现表达递减的蛋白有ATP合酶α亚单位(ATPA)、线粒体1Q补体结合蛋白(C1QBP)、组织蛋白酶D(CATD);而14-3-3ζ/δ在腺癌组中表达最低,在腺瘤组中表达最高;PI-PLC X蛋白结构域3在三组间依次递增(P<0.01)。结论正常肾上腺组织、肾上腺皮质腺瘤和肾上腺皮质癌差异表达的蛋白与能量代谢、组织结构以及一些作用蛋白酶有关,表达差异的蛋白在肾上腺皮质肿瘤中参与肿瘤的形成及侵袭作用,其可能可作为潜在的鉴别肾上腺皮质肿瘤良恶性的分子标志,成为肿瘤的生物治疗靶点。
Objective To compare the proteomics discrepancies among normal adrenal tissue, adrenal adenomas, adrenal cortical carcinoma, thereby to discover the differential diagnosis of adrenal cortical tumors and the onset of adrenal cortical carcinoma, and to investigate the transfer mechanism and therapeutic targets of adrenal cortical carcinoma. Methods Total protein was extracted from nine normal adrenal tissue, nine adrenocortical adenoma tissue samples and nine adrenocortical carcinoma samples, respectively. Immobilized pH gradient two-dimensional polyacrylamide gel eleetrophoresis was used to separate the total protein of adrenocortical adenoma tissue samples and adrenocortical carcinoma samples after protein was purified, imageMaster 2DE software was used to find out differential protein after silver staining. Then, MALDI TOF/TOF mass spectrometric identification of differential protein spots was conducted. Peptide mass fingerprints and query were achieved to find out matching proteins in the SwissProt database. Results Dielectrophoresis silver staining map had good repeatability, protein spots of normal adrenal tissue, adrenocorti- cal adenoma tissue and adrenal cortical carcinoma tissue were （2 247 ±87）, （2 375 ±142） and （2 143 ±107）, respectively. The expression of ATP synthase （ATPA）, mitochondria 1 Q complement binding protein （C1 QBP）, cathepsin D （CATD）in three groups were diminishing, while the expression of PI-PLC X protein was increased, the cancer group had the lowest expression of 14 -3 -3ζ , and the adenoma group had the highest expression. Conclusions The differentially expressed proteins among the three groups are associate with energy metabolism, organizational structure and some of the effects of protease, they are involved in tumor formation and invasion of tumors in the adrenal cortex, and may be some potential molecular markers to identify benign and malignant adrenal tumor, and become therapeutic target of cancer.
出处
《内科》
2015年第6期759-764,共6页
Internal Medicine
基金
国家自然科学基金(81060220)
广西中医药科技专项立项课题(GZKZ110-109)
