Kumarasamy Gaayathri, Ismail Mohd Nazri, Tuan Sharif Sharifah Emilia, Desire Christopher, Mittal Parul, Hoffmann Peter, Kaur Gurjeet
Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Minden 11800, Pulau Pinang, Malaysia.
Analytical Biochemistry Research Centre (ABrC), Universiti Sains Malaysia, Bayan Lepas 11900, Pulau Pinang, Malaysia.
Curr Issues Mol Biol. 2023 Apr 20;45(4):3603-3627. doi: 10.3390/cimb45040235.
Nearly 90% of cervical cancers are linked to human papillomavirus (HPV). Uncovering the protein signatures in each histological phase of cervical oncogenesis provides a path to biomarker discovery. The proteomes extracted from formalin-fixed paraffin-embedded tissues of the normal cervix, HPV16/18-associated squamous intraepithelial lesion (SIL), and squamous cell carcinoma (SCC) were compared using liquid chromatography-mass spectrometry (LC-MS). A total of 3597 proteins were identified, with 589, 550, and 1570 proteins unique to the normal cervix, SIL, and SCC groups, respectively, while 332 proteins overlapped between the three groups. In the transition from normal cervix to SIL, all 39 differentially expressed proteins were downregulated, while all 51 proteins discovered were upregulated in SIL to SCC. The binding process was the top molecular function, while chromatin silencing in the SIL vs. normal group, and nucleosome assembly in SCC vs. SIL groups was the top biological process. The PI3 kinase pathway appears crucial in initiating neoplastic transformation, while viral carcinogenesis and necroptosis are important for cell proliferation, migration, and metastasis in cervical cancer development. Annexin A2 and cornulin were selected for validation based on LC-MS results. The former was downregulated in the SIL vs. normal cervix and upregulated in the progression from SIL to SCC. In contrast, cornulin exhibited the highest expression in the normal cervix and lowest in SCC. Although other proteins, such as histones, collagen, and vimentin, were differentially expressed, their ubiquitous expression in most cells precluded further analysis. Immunohistochemical analysis of tissue microarrays found no significant difference in Annexin A2 expression between the groups. Conversely, cornulin exhibited the strongest expression in the normal cervix and lowest in SCC, supporting its role as a tumor suppressor and potential biomarker for disease progression.
近90%的宫颈癌与人乳头瘤病毒（HPV）有关。揭示宫颈癌发生各组织学阶段的蛋白质特征为生物标志物的发现提供了一条途径。使用液相色谱-质谱联用（LC-MS）比较了从正常宫颈、HPV16/18相关的鳞状上皮内病变（SIL）和鳞状细胞癌（SCC）的福尔马林固定石蜡包埋组织中提取的蛋白质组。共鉴定出3597种蛋白质，其中分别有589、550和1570种蛋白质是正常宫颈、SIL和SCC组所特有的，而三组之间有332种蛋白质重叠。在从正常宫颈到SIL的转变过程中，所有39种差异表达的蛋白质均下调，而在SIL到SCC阶段发现的所有51种蛋白质均上调。结合过程是首要的分子功能，而SIL与正常组之间的染色质沉默以及SCC与SIL组之间的核小体组装是首要的生物学过程。PI3激酶途径在启动肿瘤转化过程中似乎至关重要，而病毒致癌作用和坏死性凋亡在宫颈癌发展过程中的细胞增殖、迁移和转移中起重要作用。基于LC-MS结果选择了膜联蛋白A2和兜甲蛋白进行验证。前者在SIL与正常宫颈相比时下调，在从SIL进展到SCC时上调。相反，兜甲蛋白在正常宫颈中表达最高，在SCC中最低。尽管其他蛋白质，如组蛋白、胶原蛋白和波形蛋白也有差异表达，但它们在大多数细胞中的普遍表达妨碍了进一步分析。组织芯片的免疫组织化学分析发现，各组之间膜联蛋白A2的表达没有显著差异。相反，兜甲蛋白在正常宫颈中表达最强，在SCC中最低，支持其作为肿瘤抑制因子和疾病进展潜在生物标志物的作用。
