Effect of Pectin on the Expression of Proteins Associated with Mitochondrial Biogenesis and Cell Senescence in HT29-Human Colorectal Adenocarcinoma Cells

Abstract

Mitochondria dynamic is regulated by different proteins, maintaining a balance between fission and fusion. An imbalance towards mitochondrial fission has been associated with tumor cell proliferation. The aim of this study was to analyze whether pectin modifies the viability of human colon cancer cells and the expression of proteins involved in mitochondrial fusion and fission. The human colon carcinoma cell line HT29 cells was growth in 10% fetal bovine serum in the absence and presence of pectin. Pectin reduced HT29 cell viability in a concentration-dependent manner, reaching a plateau at 150~300 μmol/L pectin. The presence of 200 μmol/L pectin reduced the expression of dynamin-related protein-1 and increased expression of the mitochondrial fusion-associated proteins mitofusin-1 and 2. Expression of cyclin B1, a protein involved in G2/M transition, was found decreased in pectin-incubated HT29 cells. Moreover, expression of p53 protein, the amount of p53 in the nucleous and β-galactosidase activity, which are all biomarkers for cellular senescence, were significantly higher in pectin-incubated HT29 cells than in HT29 cells incubated without pectin. Expression of the protein B-cell lymphoma 2 (Bcl-2) homologous antagonist/killer was increased in response to incubation with pectin. However, incubation with pectin did not affect expression of Bcl-2-associated X protein or Bcl-2, or the caspase-3 activity. Overall, we concluded that pectin reduces the viability of human HT29 colon cancer cells, which is accompanied with a shift in the expression of proteins associated with mitochondrial dynamics towards mitochondrial fusion. Moreover, incubation with pectin favors cellular senescence over apoptosis in HT29 cells.

Cancer Stem Cells—Origins and Biomarkers: Perspectives for Targeted Personalized Therapies

“…Senescent cells are characterized by biochemical and morphological changes such as flattening and/or nuclear enlargement (508). There are several classical biomarkers of cellular senescence and they comprise: senescence-associated beta-galactosidase (SA-ß-gal) activity, expression of p53 protein, the amount of p53 in the nucleus, increase in expression of p14 (Arf), p16 (Ink4a) and p21 (Waf1), SASP, and often senescenceassociated heterochromatic foci (SAHF) (12,505,507,(509)(510)(511)(512)(513)(514)(515). Furthermore, senescent cells display low Ki67 levels and show levels of heterochromatin protein 1 (HP1) gamma ( 516), as well as di-or tri-methylated lysine 9 of histone H3 (H3K9Me2/3) and histone H2A variant (macroH2A) (505,517,518).…”

The status and trends of mitochondrial dynamics research: A global bibliometric and visualized analysis

“…Abnormal mitochondrial morphology was likewise found in senescent cells with altered expression profiles of molecules such as MFN1, MFN2, and OPA1 (Crane et al 2010 ; Marzetti et al 2017 ). Futhermore, the relevance of mitochondrial dynamics to ischemic diseases (Yu et al 2020 ; Maneechote et al 2019 ; Guan et al 2019 ), renal diseases (Zhong et al 2021 ; Chen et al 2020b ), immune-related diseases (Mills et al 2017 ; Banoth and Cassel 2018 ) and cancer (Porporato et al 2018 ; Zamorano-León et al 2019 ), among others, has been studied.In addition, the mechanism of the treatment through mitochondrial pathway for related diseases may become a popular topic in the future.…”

Mitochondrial fusion–fission dynamics and its involvement in colorectal cancer

“…Atractylenolide I, the major bioactive component in Atractylodes macrocephala , blocks DRP1‐mediated mitochondrial fission, inhibits the activation of NLRP3 inflammasome in colitis‐associated cancer (CAC), and disrupts mitochondrial membrane integrity, thereby inducing apoptosis [ 56 ]. Pectin, a polysaccharide, has been found to decrease DRP1 expression and increase mitochondrial fusion‐ and cellular senescence‐associated protein expression (e.g., β‐galactosidase and p53), thereby reducing cell viability and promoting cellular senescence [ 57 ]. Some commonly used drugs for the treatment of chronic diseases have also been reported to inhibit CRC development, possibly by modulating mitochondrial dynamics.…”