Phang Wai-Mei, Tan Aik-Aun, Gopinath Subash C B, Hashim Onn H, Kiew Lik Voon, Chen Yeng
1. Department of Oral Biology & Biomedical Sciences, Faculty of Dentistry, University of Malaya, Kuala Lumpur 50603, Malaysia.
2. Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang 11800, Malaysia.
Int J Med Sci. 2016 Apr 26;13(5):330-9. doi: 10.7150/ijms.14341. eCollection 2016.
Breast cancer is one of the most common cancers that affect women globally and accounts for ~23% of all cancers diagnosed in women. Breast cancer is also one of the leading causes of death primarily due to late stage diagnoses and a lack of effective treatments. Therefore, discovering protein expression biomarkers is mandatory for early detection and thus, critical for successful therapy. Two-dimensional electrophoresis (2D-E) coupled with lectin-based analysis followed by mass spectrometry were applied to identify potential biomarkers in the secretions of a murine mammary carcinoma cell line. Comparisons of the protein profiles of the murine 4T1 mammary carcinoma cell line and a normal murine MM3MG mammary cell line indicated that cadherin-1 (CDH), collagenase 3 (MMP-13), Viral envelope protein G7e (VEP), Gag protein (GAG) and Hypothetical protein LOC433182 (LOC) were uniquely expressed by the 4T1 cells, and pigment epithelium-derived factor (PEDF) was exclusively secreted by the MM3MG cells. Further analysis by a lectin-based study revealed that aberrant O-glycosylated CDH, N-glycosylated MMP-13 and LOC were present in the 4T1 medium. These differentially expressed N- and O-linked glycoprotein candidates, which were identified by combining lectin-based analysis with 2D-E, could serve as potential diagnostic and prognostic markers for breast cancer.
乳腺癌是全球影响女性的最常见癌症之一，约占女性确诊癌症总数的23%。乳腺癌也是主要因晚期诊断和缺乏有效治疗而导致死亡的主要原因之一。因此，发现蛋白质表达生物标志物对于早期检测至关重要，进而对于成功治疗也至关重要。二维电泳（2D-E）结合基于凝集素的分析，随后进行质谱分析，被用于鉴定小鼠乳腺癌细胞系分泌物中的潜在生物标志物。对小鼠4T1乳腺癌细胞系和正常小鼠MM3MG乳腺细胞系的蛋白质谱进行比较表明，钙黏蛋白-1（CDH）、胶原酶3（MMP-13）、病毒包膜蛋白G7e（VEP） gag蛋白（GAG）和假设蛋白LOC433182（LOC）在4T1细胞中独特表达，而色素上皮衍生因子（PEDF）仅由MM3MG细胞分泌。基于凝集素的研究进一步分析显示，4T1培养基中存在异常O-糖基化的CDH、N-糖基化的MMP-13和LOC。通过将基于凝集素的分析与2D-E相结合鉴定出的这些差异表达的N-和O-连接糖蛋白候选物，可作为乳腺癌的潜在诊断和预后标志物。
