Immunohistochemical and electron microscopic morphometric image analysis of hepatocellular carcinoma in association of HCV infection

ABSTRACT

Early detection of hepatocellular carcinoma (HCC) is crucial for successful therapy. The present work examined the value of ultrastructural morphometric image analysis of hepatocyte nuclei in patients with chronic hepatitis C virus (HCV) versus HCC cases with chronic HCV and the corresponding surgical tumor-free safe margins (TFMs), to highlight any early predictive signs of neoplastic cellular transformation. This work also performed an immunohistochemical assessment of cytokeratin 19 (CK19) and Ki-67-positive cells to visualize any associated proliferative activity in the examined groups.

The results showed significant decrease in the hepatocyte nuclear surface areas in the HCC and TFMs versus those in the HCV cases. The hepatocyte nucleolar surface area was significantly increased in the HCC cases versus that in the HCV cases. This increase was associated with a significant increase in Ki-67-positive cells in the HCC cases compared to those in the other groups. Conversely, the mean number of CK 19-positive cells was significantly reduced in the HCC cases compared to the cell numbers in TFMs and HCV cases with severe hepatic fibrosis. Liver progenitor cells (LPCs) were discerned in the reactive ductules and canaliculo-ductular junctions that characterized TFMs. LPCs were sporadically distributed in the liver lobules and reactive bile ductules in the HCC samples.

In conclusion, CK 19 represents an important marker for distinguishing between dysplastic and malignant liver nodules. Electron microscopic morphometric image analysis may be considered as adjunct factor for assessing hepatocyte malignant transformation. Wider scale studies are needed to authenticate these results.

KEYWORDS:

• HCC
• chronic HCV
• Ki-67
• liver progenitor cells
• cytokeratin 19
• ultrastructural morphometric image analysis

Acknowledgments

The authors appreciate the role of the Theodor Bilharz Research Institute in funding this work through project 111T. The authors thank Dr. Hoda Abotaleb, researcher of Biomedical Statistic at the Theodor Bilhar Research Institute, Egypt, for the statistical analysis of the obtained data.

Declaration of interest

The authors who have participated in this work declared that they do not have any conflict of interest with respect to this manuscript.

Additional information

Notes on contributors

Soheir S Mansy

Sarah Hassan collected the specimens needed for the study, performed the preliminary microscopic examinations, morphometric image analysis, and result analysis, and contributed to manuscript discussion and writing. Soheir Mansy obtained funding, developed the study concept and design, contributed to the microscopic examination, interpretation and analysis of enrolled samples, discussion of the obtained results, and manuscript editing. Ahmed S. AbdelFattah provided the study with the core liver biopsies and patient clinical data. Sahar Abdelhamid Tabak, Ahmed Mahmoud Abd El-Aziz supervision and critical review. Olfat Hamam supervised the automatic technique of immunohistochemistry. Mohammed I Seleem and Amr Abdelaa delivered the resected HCCs enrolled in the study and the patient clinical data.