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Isolation, Culture, and Characterization of Primary Dermal Fibroblasts from Human Keloid Tissue
04:41

Isolation, Culture, and Characterization of Primary Dermal Fibroblasts from Human Keloid Tissue

Published on: July 28, 2023

[Apoptosis of keloid-derived fibroblasts induced by Fas gene transfection]

Yong Luo1, Jian-hua Gao, Fei Zhao

  • 1Department of Plastic Surgery, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.

Di 1 Jun Yi Da Xue Xue Bao = Academic Journal of the First Medical College of PLA
|October 16, 2003
PubMed
Abstract

Insights

Fas gene transfection successfully induced apoptosis in keloid fibroblasts. This suggests that Fas gene mutations may contribute to keloid pathogenesis by blocking apoptosis pathways.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Dermatology

Background:

  • Keloids are abnormal scars resulting from excessive fibroblast proliferation.
  • The apoptosis pathway, particularly involving the Fas receptor, plays a crucial role in regulating cell death.
  • Dysregulation of apoptosis is implicated in the pathogenesis of various fibrotic conditions, including keloids.

Purpose of the Study:

  • To investigate the effect of Fas gene transfection on inducing apoptosis in keloid-derived fibroblasts.
  • To explore the potential role of Fas gene dysfunction in keloid formation.

Main Methods:

  • Molecular cloning was used to construct the pcDNA-3.1-Fas expression plasmid.
  • Lipofection was employed to transfect the recombinant plasmid into keloid-derived fibroblasts.
  • Apoptosis induction was achieved using Fas monoclonal antibody (mAb) and subsequently evaluated via HE staining, DNA agarose gel electrophoresis, and flow cytometry.

Main Results:

  • Successful construction of the pcDNA-3.1-Fas expression plasmid was confirmed.
  • Transfection with the Fas gene and subsequent treatment with Fas mAb induced characteristic features of apoptosis in keloid fibroblasts, including nuclear condensation, DNA laddering, and a high apoptosis rate.
  • Control cells without Fas gene transfection did not exhibit these apoptotic changes.

Conclusions:

  • The results indicate that the Fas protein is functional in inducing apoptosis in keloid fibroblasts.
  • Non-function of the Fas protein and potential Fas gene mutations may underlie the blockage of the upstream apoptosis pathway in keloid-derived fibroblasts.
  • These findings suggest that Fas gene mutations could be a contributing factor to the pathogenesis of keloids.

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