Ascites from Ovarian Cancer Induces Novel Fucosylated Proteins

Abstract

Ovarian cancer is considered to be the most lethal type of gynecological cancer. During the advanced stages of ovarian cancer, an accumulation of ascites is observed. Fucosylation has been classified as an abnormal post-translational modification that is present in many diseases, including ovarian cancer. Ovarian cancer cells that are cultured with ascites stimulation change their morphology; concomitantly, the fucosylation process is altered. However, it is not known which fucosylated proteins are modified. The goal of this work was to identify the differentially fucosylated proteins that are expressed by ovarian cancer cell lines that are cultured with ovarian cancer patients' ascites. Aleuria aurantia lectin was used to detect fucosylation, and some changes were observed, especially in the cell membrane. Affinity chromatography and mass spectrometry (MALDI-TOF) were used to identify 6 fucosylated proteins. Four proteins (Intermediate filament family orphan 1 [IFFO1], PHD finger protein 20-like protein 1 [PHF20L1], immunoglobulin gamma 1 heavy chain variable region partial [IGHV1-2], and Zinc finger protein 224 [ZNF224]) were obtained from cell cultures stimulated with ascites, and the other two proteins (Peregrin [BRPF1] and Dystrobrevin alpha [DTNA]) were obtained under normal culture conditions. The fucosylated state of some of these proteins was further analyzed. The experimental results show that the ascites of ovarian cancer patients modulated the fucosylation process. The PHD finger protein 20-like protein 1, Zinc finger protein 224 and Peregrin proteins colocalize with fucosylation at different levels.

Overexpression of Fut 2, 4, and 8, and nuclear localization of Fut 4 in ovarian cancer cell lines induced by ascitic fluids from epithelial ovarian cancer patients

“…In the case of OVCAR-3 and CAOV-3, the effect of the five AFs did not generate structural changes in the remodeling of the cytoskeleton or in the morphology of these cells; however, the cells maintained high levels of fucosylation around the cell membrane, as we previously reported for SKOV-3 and OVCAR-3 cells (Alberto-Aguilar et al, 2019). The rounded cellular morphology of OVCAR-3 and CAOV-3 suggests that their migration could be anchorageindependent, since the cells present in ovarian tumor spheroids have a high potential for proliferation and migration (Jianqun et al, 2014;Wang et al, 2012).…”

“…Cells (SKOV‐3, OVCAR‐3, and CAOV‐3) were processed as described (Alberto‐Aguilar et al, 2019), for the analysis of total fucosylation using a biotinylated Aleuria aurantia lectin (Vector Laboratories, B‐1395, 1:50 dilution), and a streptavidin‐FITC conjugate (Life Technologies, 4343‐11, 1:100 dilution). For the detection of FUTs, the primary antibodies were incubated in a 1:50 dilution in PBS overnight at 4°C: anti‐Fut 2 (sc‐21964), anti‐Fut 4 (sc‐292247), and anti‐Fut 8 (sc‐271244) all obtained from Santa Cruz Biotechnology Inc.…”

“…Cells (SKOV-3, OVCAR-3, and CAOV-3) were processed as described (Alberto-Aguilar et al, 2019), for the analysis of total fucosylation…”

ZEB2 facilitates peritoneal metastasis by regulating the invasiveness and tumorigenesis of cancer stem-like cells in high-grade serous ovarian cancers

“…Malignant ascites is an important clinical outcome of HGSOC invasion [ 28 – 30 ] and promotes the dissemination of cancer cells from the primary tumour mass to the peritoneal cavity. However, intraperitoneal metastasis is not a simple tumour cell shedding and dissemination.…”

Identification of bromodomain-containing proteins prognostic value and expression significance based on a genomic landscape analysis of ovarian serous cystadenocarcinoma

“…Alberto-Aguilar DR et al. found that the ascites of OSC patients could modulate the fucosylation of BRPF1 to promote OSC development ( 93 ). These results suggested that BRPF1, as a highly conserved oncogene, might play equally important oncogenic roles in different cancers.…”