Effect of apelin on mitosis, apoptosis and DNA repair enzyme OGG 1/2 expression in intestinal cell lines IEC-6 and Caco-2

Abstract

Apelin is a regulatory peptide, identified as an endogenous ligand of the Apelin receptor (APJ). Both the apelin and the APJ were detected in brain, lung, heart, mammary gland, kidney, placenta, adipose tissues and the gastrointestinal tract. Apelin is considered an important regulatory gut peptide with a potential physiological role in gastrointestinal cytoprotection, regulation of food intake and drinking behaviour. The aim of the present study was to assess the effect of the apelin on mitosis, apoptosis and the expression of DNA repair enzyme (OGG 1/2), and APJ receptor in intestinal cell lines: rat crypt (IEC-6) and human enterocyte model (Caco-2). The cell cultures were incubated with the apelin-12 (10 -8 M) for 4, 6, 12, 24 and 48 h and the apoptosis (caspase 3), mitosis (Ki-67) and DNA repair enzyme (OGG1/2) markers were studied by Real-Time qRT-PCR and immunofluorescent methods. The results of Real-Time qRT-PCR and immunocytochemical analysis showed that the levels of mRNAs were inversely related to the expression level of corresponding proteins. Immunofluorescent studies revealed inhibitory effect of apelin-12 on apoptosis, mitosis and the expression of OGG1/2 in the intestinal crypt cell line IEC-6. However, in the enterocyte model Caco-2 cells apelin stimulated apoptosis and mitosis, and reduced OGG1/2 expression. These findings suggest that apelin may be involved in the control of epithelial cell turnover in the gastrointestinal tract.

In Vitro Influence of Extracts from Snail Helix aspersa Müller on the Colon Cancer Cell Line Caco-2

“…Human epithelial colorectal adenocarcinoma (Caco-2) cell line (ECCC, 55 passage; Sigma-Aldrich, St. Louis, MO, USA) was grown in 96-well plastic plates (at a density of 1 × 10 4 cells/100 µL) in Minimum Essential Medium (MEM) containing 2 mM l -glutamine (Thermo Fisher Scientific, Waltham, MA, USA), with 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Waltham, MA, USA), 1% non-essential amino acids (NEAA, Thermo Fisher Scientific, Waltham, MA, USA) and 1% antibiotic-antimycotic (Thermo Fisher Scientific, Waltham, MA, USA) [ 79 ]. The cultures were kept at 37 °C in 5% CO 2 and 95% relative humidity (CO 2 incubator INCO 108 med, Memmert GmbH + Co. KG, Schwabach, Germany).…”

Apelin's effects on young rat gastrointestinal tract maturation

“…Those in vitro studies showed that apelin suppressed apoptosis, mitosis and OGG 1,2 expression in the intestinal crypt cell line IEC-6 but stimulated apoptosis and mitosis in the Caco-2 cells line. Apelin also suppressed OGG 1,2 expression in Caco-2 cells, suggesting that apelin may be involved in gastrointestinal tract cell turnover [1]. It was previously found that apelin stimulates [15,17,18] or inhibits [29] cell proliferation, depending on the context.…”

Apelin Effects Migration and Invasion Abilities of Colon Cancer Cells

“…Blebs formation is often connected with apoptosis [ 36 ]. According to many studies apelin may prevent apoptosis [ 37 , 38 ]; therefore, we examined if apelin could prevent staurosporin (STS)-induced apoptosis of colon cancer cells using Annexin V-FITC and propidium iodide assay ( Supplementary Figure 3 ). Our results showed no ability of apelin to prevent STS-induced apoptosis, as well as no pro-apoptotic effect due to the fact that the apoptotic process has not been enhanced by apelin.…”