Membrane proteomics of cervical cancer cell lines reveal insights on the process of cervical carcinogenesis

Abstract

The available therapeutic approaches for cervical cancer can seriously affect the fertility potential of patient; thus, there is a pressing requirement for less toxic and targeted therapies. The membrane proteome is a potential source of therapeutic targets; however, despite the significance of membrane proteins in cancer, proteomic analysis has been a challenging task due to their unique biochemical properties. The aim of the present study was to develop an efficient membrane protein enrichment protocol, and to the best of our knowledge, to compare for the first time the expression pattern of membrane proteins of one normal cell line, HCK1T, and three cervical cancer cell lines, C33A, a human papilloma virus (HPV)-negative cell line, and two HPV-positive cell lines, SiHa (HPV16+) and HeLa (HPV18+). The study aimed to identify the proteins that are involved in cervical carcinogenesis and may constitute novel drug targets. Membrane protein isolation, liquid chromatography coupled with tandem mass spectrometry proteomics and bioinformatics analysis were performed in the membrane fraction of the informative cervical cell lines following a novel enrichment protocol. The percentages of membrane and transmembrane proteins in the enrichment protocol were higher compared with those of the corresponding data derived from total cell extract analysis. Differentially expressed proteins were detected by the comparison of the cervical cancer cell lines with the normal cell line. These proteins constitute molecular features of cancer pathology and participate in biological pathways relevant to malignancy, including 'HIPPO signaling', 'PI3K/Akt signaling', 'cell cycle: G2/M DNA damage checkpoint regulation' and 'EIF2 signaling'. These unique membrane protein identifications offer insights on a previously inaccessible region of the cervical cancer proteome, and may represent putative diagnostic and prognostic markers, and eventually therapeutic targets.

High resolution analysis of the intracellular proteome of cervical cancer cell lines unveils novel regulators of cervical carcinogenesis

“…A need for deeper understanding of the underlying mechanisms of cervical cancer pathology is critical. Towards this goal, our group has already investigated the secretome (9) and the membrane proteomes (10) of three cervical cancer cell lines in comparison to normal cervical keratinocytes with liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). These studies highlighted the inhibition of matrix metalloproteases (9) and the deregulation of HIPPO, PI3K/Akt, and EIF2 signaling pathways (10) in cervical cancer as well as many differentially expressed proteins that are potentially crucial for cervical malignancy.…”

“…Towards this goal, our group has already investigated the secretome (9) and the membrane proteomes (10) of three cervical cancer cell lines in comparison to normal cervical keratinocytes with liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). These studies highlighted the inhibition of matrix metalloproteases (9) and the deregulation of HIPPO, PI3K/Akt, and EIF2 signaling pathways (10) in cervical cancer as well as many differentially expressed proteins that are potentially crucial for cervical malignancy. Along the same lines and in an attempt to elucidate the molecular mechanisms underlining cervical carcinogenesis, we analyzed the intracellular proteome of three cervical cancer cell lines against normal cervical keratinocytes employing high resolution LC-MS/MS.…”

Understanding Cervical Cancer through Proteomics

“…The Mann-Whitney statistical analysis used to compare the results with the non-cancer cell line HCK1T allowed them to identify 263 unique transmembrane proteins in C33, 262 unique transmembrane proteins in HeLa, and 152 unique transmembrane proteins in SiHa. Among the identified transmembrane proteins, TMX2, FAM120A, CLPTM1, CKAP5, and NCSTN were the most prominent proteins differentially expressed in CC cell lines [105].…”

“…Another strategy for studying CC based on proteomics is the study of cell membranes as possible therapeutic targets. In 2018, Pappa et al [105] published a work on the isolation and enrichment of membrane proteins of three different cell lines, HeLa, SiHa, and C33A CC. They performed a proteomic characterization of these cell lines by LC-MS/MS and a bioinformatics analysis using Proteome Discoverer 1.4, SEQUEST, and UniProt.…”

Proteomic analysis of human cervical adenocarcinoma mucus to identify potential protein biomarkers

“…Proteomics analysis is a powerful tool for monitoring the change of protein levels to discover new biomarkers in many cancers, such as colorectal cancer, pancreatic cancer, and neuroendocrine cervical cancer ( Lin et al, 2014 ; Pan, Brentnall & Chen, 2015 ; Chauvin & Boisvert, 2018 ). Unique membrane proteins have been identified using proteomics analysis of the cervical cancer cell lines ( Pappa et al, 2018 ). Lee et al (2011) studied the proteome of cervical mucus plugs and suggested its role for maintaining pregnancy and parturition.…”