Negative Correlation between Cytoglobin Expression and Intracellular ROS Levels in Human Skin Keloid Fibroblasts

Fajri Marindra Siregar, Novi Silvia Hardiany, Sri Widia Azraki Jusman


BACKGROUND: In our previous study, we found higher cytoglobin (Cygb) expression in keloid than normal tissue. Cytoglobin is a new globin family protein which function is still being studied to date. The purpose of this research is to elucidate the function of Cygb in human skin keloid fibroblasts (KFs), especially its role in intracellular reactive oxygen species (ROS) levels.

METHODS: The study was conducted on human skin KFs obtained from primary culture. Inhibition of Cygb expression was achieved by using siRNA targeting Cygb. We compared the relative expression of Cygb between treatment and control group, and its effect on intracellular ROS levels. Gene expression was measured using quantitative real-time polymerase chain reaction (qRT-PCR) while the ROS level counted by dichlorodihydrofluorescein diacetate (DCFHDA) assay.

RESULTS: There was an increase in intracellular ROS levels in the small interfering RNA (siRNA) (+) Cygb group compared to control group (1.673 vs. 1.260; 1.773 vs. 1.393; 1.710 vs. 1.360; respectively). There is a negative correlation between Cygb expression and ROS level (p<0.05; r=-0.651).

CONCLUSION: There is a negative correlation between Cygb expression and intracellular ROS levels, we suggest Cygb acts as a ROS scavenger in human skin KFs.

KEYWORDS: skin keloid fibroblasts, cytoglobin, siRNA, ROS

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Zhang Q, Yamaza T, Kelly AP, Shi S, Wang S, Brown J, et al. Tumorlike stem cells derived from human keloid are governed by the inflammatory niche driven by IL-17/IL-6 axis. PloS One. 2009; 4: e7798, CrossRef.

Wolfram D, Tzankov A, Pülzl P, Piza-Katzer H. Hypertrophic scars and keloids--a review of their pathophysiology, risk factors, and therapeutic management. Dermatol Surg Off Publ Am Soc Dermatol Surg Al. 2009; 35: 171-81, CrossRef.

Gragnani A, Warde M, Furtado F, Ferreira LM. Topical tamoxifen therapy in hypertrophic scars or keloids in burns. Arch Dermatol Res. 2010; 302: 1-4, CrossRef.

Xue M, Jackson CJ. Extracellular matrix reorganization during wound healing and its impact on abnormal scarring. Adv Wound Care. 2015; 4: 119-36, CrossRef.

Zhang Q, Wu Y, Chau CH, Ann DK, Bertolami CN, Le AD. Crosstalk of hypoxia-mediated signaling pathways in upregulating plasminogen activator inhibitor-1 expression in keloid fibroblasts. J Cell Physiol. 2004; 199: 89-97, CrossRef.

Wulandari E. The Role of Cytoglobin in Fibrosis Hypoxia with Keloid as A Model [Dissertation]. Jakarta: Universitas Indonesia; 2016.

De Felice B, Wilson RR, Nacca M. Telomere shortening may be associated with human keloids. BMC Med Genet. 2009; 10: 110, CrossRef.

Shaw RJ, Omar MM, Rokadiya S, Kogera FA, Lowe D, Hall GL, et al. Cytoglobin is upregulated by tumour hypoxia and silenced by promoter hypermethylation in head and neck cancer. Br J Cancer. 2009; 101: 139-44, CrossRef.

Jusman SWA, Iswanti FC, Suyatna FD, Ferdinal F, Wanandi SI, Sadikin M. Cytoglobin expression in oxidative stressed liver during systemic chronic normobaric hypoxia and relation with HIF-1α. Med J Indones. 2014; 23: 133-8, CrossRef.

Singh S, Manda SM, Sikder D, Birrer MJ, Rothermel BA, Garry DJ, et al. Calcineurin activates cytoglobin transcription in hypoxic myocytes. J Biol Chem. 2009; 284: 10409-21, CrossRef.

Mimura I, Nangaku M, Nishi H, Inagi R, Tanaka T, Fujita T. Cytoglobin, a novel globin, plays an antifibrotic role in the kidney. Am J Physiol Renal Physiol. 2010; 299: F1120-33, CrossRef.

Sari DH. Role of Hypoxia Inducible Factor-1alpha (HIF-1α) and Hypoxia Inducible Factor-2alpha (HIF-2α) to Cytoglobin Expression and Fibroblast Proliferation on Keloid [Thesis]. Jakarta: Universitas Indonesia; 2017.

Bholah TC, Neergheen-Bhujun VS, Hodges NJ, Dyall SD, Bahorun T. Cytoglobin as a biomarker in cancer: potential perspective for diagnosis and management. BioMed Res Int. 2015; 2015: 1-6, CrossRef.

Singh S, Canseco DC, Manda SM, Shelton JM, Chirumamilla RR, Goetsch SC, et al. Cytoglobin modulates myogenic progenitor cell viability and muscle regeneration. Proc Natl Acad Sci USA. 2014; 111: E129-38, CrossRef.

Oleksiewicz U, Liloglou T, Field JK, Xinarianos G. Cytoglobin: biochemical, functional and clinical perspective of the newest member of the globin family. Cell Mol Life Sci CMLS. 2011; 68: 3869-83, CrossRef.

Rao DD, Vorhies JS, Senzer N, Nemunaitis J. siRNA vs. shRNA: similarities and differences. Adv Drug Deliv Rev. 2009; 61: 746-59, CrossRef.

Thuy LTT, Matsumoto Y, Thuy TTV, Hai H, Suoh M, Urahara Y, et al. Cytoglobin deficiency promotes liver cancer development from hepatosteatosis through activation of the oxidative stress pathway. Am J Pathol. 2015; 185: 1045-60, CrossRef.

McRonald FE, Risk JM, Hodges NJ. Protection from intracellular oxidative stress by cytoglobin in normal and cancerous oesophageal cells. PloS One. 2012; 7: e30587, CrossRef.

Fordel E, Thijs L, Moens L, Dewilde S. Neuroglobin and cytoglobin expression in mice. Evidence for a correlation with reactive oxygen species scavenging. FEBS J. 2007; 274: 1312-7, CrossRef.


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