Markers of Angiogenesis, Lymphangiogenesis, and Epithelial–Mesenchymal Transition (Plasticity) in CIN and Early Invasive Carcinoma of the Cervix: Exploring Putative Molecular Mechanisms Involved in Early Tumor Invasion

Abstract

The establishment of a proangiogenic phenotype and epithelial-to-mesenchymal transition (EMT) are considered as critical events that promote the induction of invasive growth in epithelial tumors, and stimulation of lymphangiogenesis is believed to confer the capacity for early dissemination to cancer cells. Recent research has revealed substantial interdependence between these processes at the molecular level as they rely on common signaling networks. Of great interest are the molecular mechanisms of (lymph-)angiogenesis and EMT associated with the earliest stages of transition from intraepithelial development to invasive growth, as they could provide the source of potentially valuable tools for targeting tumor metastasis. However, in the case of early-stage cervical cancer, the players of (lymph-)angiogenesis and EMT processes still remain substantially uncharacterized. In this study, we used RNA sequencing to compare transcriptomes of HPV(+) preinvasive neoplastic lesions and early-stage invasive carcinoma of the cervix and to identify (lymph-)angiogenesis- and EMT-related genes and pathways that may underlie early acquisition of invasive phenotype and metastatic properties by cervical cancer cells. Second, we applied flow cytometric analysis to evaluate the expression of three key lymphangiogenesis/EMT markers (VEGFR3, MET, and SLUG) in epithelial cells derived from enzymatically treated tissue specimens. Overall, among 201 differentially expressed genes, a considerable number of (lymph-)angiogenesis and EMT regulatory factors were identified, including genes encoding cytokines, growth factor receptors, transcription factors, and adhesion molecules. Pathway analysis confirmed enrichment for angiogenesis, epithelial differentiation, and cell guidance pathways at transition from intraepithelial neoplasia to invasive carcinoma and suggested immune-regulatory/inflammatory pathways to be implicated in initiation of invasive growth of cervical cancer. Flow cytometry showed cell phenotype-specific expression pattern for VEGFR3, MET, and SLUG and revealed correlation with the amount of tumor-infiltrating lymphocytes at the early stages of cervical cancer progression. Taken together, these results extend our understanding of driving forces of angiogenesis and metastasis in HPV-associated cervical cancer and may be useful for developing new treatments.

Biodegradable Scaffolds for Vascular Regeneration Based on Electrospun Poly(L-Lactide-co-Glycolide)/Poly(Isosorbide Sebacate) Fibers

“…Genotyping of HUVEC cells collected after 48 h of incubations on PISEB, PLGA and PLGA/PISEB showed a potentially pro-angiogenic expression profile. Genes of neo-angiogenesis were observed to be in the cross-talk and correlations, and a feedback loop for upregulated MMPs, together with downregulated TIMP inhibitors, resulted in tissues with invasive and migratory cell phenotypes [ 41 , 42 ]. Under those conditions, cells cultured on the surface of PLGA/PISEB changed morphology into that typical for tube-formation specimens during neo-angiogenesis process [ 30 ].…”

“…After cells were trypsinized and collected, total RNA was isolated using the phenol-chloroform method of extraction, by the Total RNA Isolation kit (A@A Biotechnology, Gdańsk, Poland). The efficiency of RNA isolation was assessed spectrophotometrically, and amplification of genes was performed using commercially-available kits (Real-Time 2xPCR Master Mix SYBR A; A@A Biotechnology) and set of primers for IL8, VEGF, MMP2, MMP9, TIMP1 and TIMP2 (Genomed, Warsaw, Poland) from references [ 41 , 42 , 52 ]. The quantitative PCR reaction, preceded by reverse transcription (NG dART RT kit, EURx, Gdańsk, Poland), was performed using a CFX96 Touch™ Real-Time PCR Detection System thermocycler (Bio-Rad, Hercules, CA, USA).…”

Retinal aging transcriptome and cellular landscape in association with the progression of age-related macular degeneration

“…This result is consistent with a study that showed increased angiogenesis during fracture healing in a MATN1 deficient mouse, and purified MATN1 proteins enabled inhibition of angiogenesis both in vitro and in vivo 28 . Of note, VSTM4, FBLN5, MPZL3 and SCPEP1, have been reported to promote sprouting angiogenesis by endothelial cells 29,30,31,32 , while their role in retinal angiogenesis remains unstudied.…”

Long noncoding RNA TUG1 upregulates VEGFA to enhance malignant behaviors in stomach adenocarcinoma by sponging miR‐29c‐3p

“…Therefore, inhibiting angiogenesis has become one of the effective strategies in cancer therapy 15 . Numerous signaling molecules, such as VEGFA and TGF‐β, have been proven to modulate angiogenesis during cancer progression over the past decades 16,17 . Many studies reported that TUG1 is involved in angiogenesis during the progression of various diseases 18,19 .…”

“… 15 Numerous signaling molecules, such as VEGFA and TGF‐β, have been proven to modulate angiogenesis during cancer progression over the past decades. 16 , 17 Many studies reported that TUG1 is involved in angiogenesis during the progression of various diseases. 18 , 19 One miRNA in particular, miR‐29c‐3p, could be directly targeted by TUG1 to stimulate angiogenesis in endothelial progenitor cells and diabetic mouse ischemic limb.…”