Wasniewski Tomasz, Kiezun Jacek, Krazinski Bartlomiej E, Kowalczyk Anna E, Szostak Blazej, Wierzbicki Piotr M, Kiewisz Jolanta
Department of Obstetrics, Perinatology and Gynecology, School of Medicine, Collegium Medicum, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.
Department of Human Histology and Embryology, School of Medicine, Collegium Medicum, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.
Folia Histochem Cytobiol. 2019;57(2):84-93. doi: 10.5603/FHC.a2019.0010. Epub 2019 Jun 14.
WNT5A (Wnt family member 5A) belongs to the WNT family of secreted signaling glycoproteins that play essential role in developmental, physiological and pathological processes. WNT5A was shown to take part in carcinogenesis process playing both oncogenic and suppressor functions in various types of human malignancies. This study aimed to assess the expression of the WNT5A gene at the mRNA and protein levels in the specimens derived from endometrial cancer (EC) or unchanged control endometrium. The associations between the WNT5A expression levels and clinicopathological characteristics and survival of EC patients were evaluated.
Total RNA was isolated in order to assess the relative amounts of WNT5A mRNA by quantitative polymerase chain reaction (QPCR) in samples of unchanged endometrial control (n = 8) and tumor samples of EC patients (n = 28). Immunohistochemistry (IHC) was used to determine the presence of WNT5A protein in the sections of formalin-fixed, paraffin-embedded tissue specimens derived from unchanged endome-trial controls (n = 6) and EC tumors (n = 19). Significance of differences in WNT5A expression levels between the studied groups of EC patients and correlations between the WNT5A and demographic data, pathological features, hematological parameters and overall survival of the patients were evaluated by statistical analysis.
The level of WNT5A mRNA was decreased in EC in comparison to unchanged endometrium. WNT5A expression was associated with primary tumor invasion status exhibiting reduced level of transcripts in EC that involved organs beyond the uterus when compared to the uterus-confined cancers. WNT5A immunoreactivity was visualized in the cytoplasm and nuclei of EC cells as well as in the luminal and glandular epithelial cells of unchanged endometrium. WNT5A mRNA expression levels correlated negatively with cytoplasmic, and positively with nuclear immunoreactivity of the WNT5A protein in the EC cells. In addition, the relationships between blood leucocyte count (in particular granulocytes and lymphocytes) of patients with EC and their WNT5A mRNA and protein expression levels were established. A positive correlation between the nuclear immunoexpression of WNT5A protein in the cancer cells in cell nuclei and mean platelet volume in blood was also found.
The results of the first study of WNT5A expression at the transcript and protein levels indicate that it could be considered as a potential marker of molecular changes that take place during EC development.
WNT5A(Wnt家族成员5A)属于分泌型信号糖蛋白的WNT家族,在发育、生理和病理过程中发挥着重要作用。WNT5A已被证明参与致癌过程,在各种类型的人类恶性肿瘤中发挥致癌和抑制功能。本研究旨在评估WNT5A基因在子宫内膜癌(EC)或未改变的对照子宫内膜标本中的mRNA和蛋白质水平表达。评估了WNT5A表达水平与EC患者临床病理特征及生存之间的关联。
提取总RNA,通过定量聚合酶链反应(QPCR)评估未改变的子宫内膜对照样本(n = 8)和EC患者肿瘤样本(n = 28)中WNT5A mRNA的相对含量。免疫组织化学(IHC)用于确定WNT5A蛋白在福尔马林固定、石蜡包埋的组织标本切片中的存在情况,这些标本来自未改变的子宫内膜对照(n = 6)和EC肿瘤(n = 19)。通过统计分析评估EC患者研究组中WNT5A表达水平差异的显著性以及WNT5A与人口统计学数据、病理特征、血液学参数和患者总生存之间的相关性。
与未改变的子宫内膜相比,EC中WNT5A mRNA水平降低。WNT5A表达与原发性肿瘤侵袭状态相关,与局限于子宫的癌症相比,累及子宫以外器官的EC中WNT5A转录本水平降低。在EC细胞的细胞质和细胞核以及未改变的子宫内膜的腔上皮和腺上皮细胞中均可观察到WNT5A免疫反应性。EC细胞中WNT5A mRNA表达水平与细胞质呈负相关,与WNT5A蛋白的核免疫反应性呈正相关。此外,还建立了EC患者的血液白细胞计数(特别是粒细胞和淋巴细胞)与其WNT5A mRNA和蛋白表达水平之间的关系。还发现癌细胞细胞核中WNT5A蛋白的核免疫表达与血液中平均血小板体积呈正相关。
首次对WNT5A转录本和蛋白水平表达的研究结果表明,它可被视为EC发生过程中分子变化的潜在标志物。