3.

International Anatolian Scientific Research Congress 2022

Kayseri , Turkiye

Abnormal observations of hepatocyte behaviours during liver

regeneration: Perspectives from Proliferating Cell Nuclear Antigen
(PCNA)
Presented by Muhammed Fawaz Abdullah

Muhammed Fawaz Abdullah1a, Mediha Canbek2b, Ayse Ozmen Yaylaci3c, Neda Hamid1d
1
Department of Biology, Graduate School of Natural and Applied Sciences, Eskisehir Osmangazi University,
26480 Eskisehir, Turkey
Department of Biology, Faculty of Arts and Science, Eskisehir Osmangazi University, 26480 Eskisehir, Turkey
2

3
Department of Biology, Faculty of Arts and Science, Hitit University, Corum, Turkey
a: orcid.org/0000-0002-7443-3031; b: orcid.org/0000-0003-1095-2382; c: orcid.org/0000-0001-9658-3316; d:
orcid.org/0000-0001-7602-2351
 LIVER REGENERATION

 Highly-regulated and well-orchestrated process of

restoring the loss of a functional liver mass after partial
hepatectomy or toxin-induced liver damage (Fausto et
al., 2006).

 A rapid and well-synchronized passage of the quiescent

G0 phase hepatocytes into the cell cycle (G1-S-G2-M).

 Epithelial tissues (intestines and skin) with high

cell turnover renew using tissue stem cells. The
liver due to its low cell, renews using all mature
hepatocytes (Gilgenkrantz and Collin, 2011;
Michalopoulos, 2017).

 Rodent model of two-thirds partial hepatectomy

(PHx) (Higgins & Anderson, 1931)

Gilgenkrantz & Collin de l'Hortet, 2018

 METABOLIC ZONATION OF LIVER REGENERATIVE LOBE

Periportal zone 1 Centrilobular zone 3

Intermediate zone 2
Periportal gluconeogenesis glycolysis Central
Tbx3, Axin2
Vein & lipogenesis Vein
&Wnt/b-catenin
Beta-oxidation detoxification

Metabolically, the lobule is the functional unit of the liver

(Gilgenkrantz & Collin de l'Hortet, 2018)
Cells involved in regeneration (Schematic
Representation).

Axin 2-positive hepatocytes

CV (pale green) ---> mediolobular (green-blue) or periportal
(pale blue)
 mild acute damage - nondamaged hepatocytes enter into
the cell cycle.
 Mild chronic damage - hybrid hepatocytes portal vein (pale
blue)
 Severe chronic liver damage - ductular reaction

Zone 1 - periportal region

Zone 2 - intermediate region
zone 3 - centrilobular region
CV - central vein
PV - portal vein.

(Gilgenkrantz & Collin de l'Hortet, 2018)

 PCNA (cyclin)

36 KDa nuclear protein, very conserved protein and known to act as an auxiliary protein for DNA polymerase delta
(Bravo et al., 1987), playing a key roles in dual regulation of DNA synthesis and cell proliferation (Takasaki et al., 1981)


PCNA as a central protein in both DNA replication and repair, encircles double-stranded DNA as a trimer, forming a

sliding clamp that tethers proteins such as polymerases to DNA (Ellison & Stillman, 2003).


PCNA is essential not only for DNA replication but also for several forms of DNA repair, including nucleotide excision
repair (NER), the major pathway by which cells remove DNA damage introduced by UV light and a variety of chemical
carcinogens (Hoogervorst et. al., 2005).
 Identified as the polymerase-associated protein and synthesized in early G1 and S phases of the cell cycle, PCNA

expression usually peaks at growth phase 1-synthesis phase (G1/S) before dropping to very low levels during mitosis,

undetectable by the immunohistochemical method (Essers et. al., 2005).

Early S phase - (very granular distribution & absent in nucleoli) vs

Late S phase - (prominent in nucleoli)

Cell fixation; organic solvents (PCNA associated with Nuclear region) vs

Aldehydes (more diffuse but intense and occurs throughout the cell cycle)

2 basic forms (PCNA protein): organic fixation-sensitive soluble form (non-replicative activity) vs insoluble with (DNA
synthesis-associated activity)

Thus, PCNA serves as an endogenous histologic marker for the G1/S phases of the cell cycle.
Cytokinesis-Block Micronucleus Assay Adapted for Analyzing Genomic Instability of Human Mesenchymal Stem Cells, 2014
(Déborah Afonso et. al. 2014)
• Binucleated cells analyzed by the cytokinesis-block micronucleus
(CBMN) assay; micronucleus (MN), nucleoplasmic bridges (NPBs),
and nuclear buds (NBUDs)were observed in the three primary hMSC
cultures and cell lines A549 and XP4PA.
• (A, E, I, M, Q) - The normal binucleated cells observed in A549,
XP4PA, hMSC1, hMSC2, and hMSC3, respectively.
• (B, F, J, N, R) - Binucleated cells with NPBs observed in A549,
XP4PA, hMSC1, hMSC2, and hMSC3, respectively.
• (C, G, K, O, S) - Binucleated cells with NBUDs observed in A549,
XP4PA, hMSC1, hMSC2, andhMSC3, respectively.
• (D, H, L, P, T) - Binucleated cells with MN observed in A549, XP4PA,
hMSC1, hMSC2, and hMSC3, respectively

A549 – Human lung cancer,

XP4PA – Human Fetus
Xeroderma pigmentosum
 Comparative analysis of anaphase bridge in 2 different cells

↖︎

A B

Anaphase bridge in hepatocyte at PH 48 hrs Anaphase bridge in liver of the subject mother mouse exposed
to DES (Göze & Çolak,1996)
Nucleoplasmic-like bridge during uncompleted cytokinesis
(PH48 hrs)

↖︎
Multi-nucleated giant hepatocyte (MNGHs) Prophase of MNGHs
PH48 hrs PH48 hrs

↖︎
↖ Extra Polyploid and ‘‘kiss & Run’’ behaviours of Hepatocytes

↖︎
Nuclear Fusion: positive (Red cell) + negative (blue cell) ↖︎Nuclear Fusion (Positive cells) & Anaphase
PH48 hrs PH72 hrs

↖︎

↖︎
 REFERENCES
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