Huang Haiyun, Hao Zhixiang, Long Lingyan, Yin Zeyuan, Wu Chenyu, Zhou Xueyan, Zhang Bei
Department of Obstetrics and Gynaecology, Xuzhou Central Hospital, Affiliated Xuzhou Clinical College of Xuzhou Medical University, Xuzhou, Jiangsu 221004, P.R. China.
Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, Jiangsu 221004, P.R. China.
Oncol Lett. 2021 May;21(5):408. doi: 10.3892/ol.2021.12669. Epub 2021 Mar 22.
The present study aimed to determine the differential expression profiles of proteins in endometrial carcinoma and to screen the proteins associated with the occurrence and development of endometrial cancer (EC). In total, 15 samples of human EC and paracancerous tissues were selected for proteomic analysis using a label-free quantification method based on liquid chromatography-tandem mass spectrometry. The differential proteins were analysed using bioinformatics and verified using reverse transcription-quantitative PCR (RT-qPCR) and western blotting. Finally, the expression of differential proteins in 75 endometrial carcinoma samples and 30 normal endometrial tissue samples were detected using immunohistochemical staining, and the associations between differential protein expression and clinicopathological features were analysed. In total, 579 up-regulated proteins and 346 down-regulated proteins were identified between the two groups and seven proteins with the most significant differences were selected; these proteins included interferon-induced protein with tetratricopeptide repeats 3, poly(ADP-ribose) polymerase family member 9, solute carrier family 34 member 2, cytochrome b5 reductase 1, protein tyrosine phosphatase non-receptor type 1, dermatopontin (DPT) and secretory leukocyte peptidase inhibitor. RT-qPCR and western blotting showed that DPT expression was down-regulated (P<0.001), which was consistent with the mass spectrometry results. The immunohistochemical staining results showed that the positive expression of DPT in EC and normal endometrial tissues was statistically significant (P<0.001). The positive expression of DPT was significantly decreased in poorly differentiated, late stage, lymph node metastasis and myometrial invasion depth ≥1/2 samples (P<0.05). DPT expression was significantly lower in EC, which might play role in the pathogenesis of EC.
本研究旨在确定子宫内膜癌中蛋白质的差异表达谱，并筛选与子宫内膜癌（EC）发生发展相关的蛋白质。总共选取了15例人EC组织和癌旁组织样本，采用基于液相色谱 - 串联质谱的无标记定量方法进行蛋白质组学分析。使用生物信息学分析差异蛋白，并通过逆转录定量PCR（RT-qPCR）和蛋白质印迹法进行验证。最后，采用免疫组织化学染色检测75例子宫内膜癌样本和30例正常子宫内膜组织样本中差异蛋白的表达，并分析差异蛋白表达与临床病理特征之间的关联。两组之间共鉴定出579个上调蛋白和346个下调蛋白，并选取了7个差异最显著的蛋白；这些蛋白包括含四肽重复序列的干扰素诱导蛋白3、聚（ADP - 核糖）聚合酶家族成员9、溶质载体家族34成员2、细胞色素b5还原酶1、非受体型蛋白酪氨酸磷酸酶1、皮肤桥蛋白（DPT）和分泌型白细胞蛋白酶抑制剂。RT-qPCR和蛋白质印迹法显示DPT表达下调（P<0.001），这与质谱结果一致。免疫组织化学染色结果显示，DPT在EC组织和正常子宫内膜组织中的阳性表达具有统计学意义（P<0.001）。在低分化、晚期、有淋巴结转移和肌层浸润深度≥1/2的样本中，DPT的阳性表达显著降低（P<0.05）。DPT在EC中的表达显著降低，可能在EC的发病机制中起作用。
