Hsp70 Knockdown by siRNA Decreased Collagen Production in Keloid Fibroblasts

There are currently no consistently effective treatments for the excessive collagen produced by keloid fibroblasts. Previously, we reported that heat shock protein 70 (Hsp70) is up-regulated in keloid fibroblasts and keloid tissue. We, therefore, investigated whether Hsp70 is related to excessive co...

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Published inYonsei medical journal Vol. 56; no. 6; pp. 1619 - 1626
Main Authors Shin, Jung U, Lee, Won Jai, Tran, Thanh-Nga, Jung, Inhee, Lee, Ju Hee
Format Journal Article
LanguageEnglish
Published Korea (South) Yonsei University College of Medicine 01.11.2015
연세대학교의과대학
Subjects
Adolescent
Adult
Blotting, Western
Collagen - drug effects
Collagen - metabolism
Female
Fibroblasts - metabolism
Gene Expression Regulation
HSP70 Heat-Shock Proteins - genetics
HSP70 Heat-Shock Proteins - metabolism
HSP70 Heat-Shock Proteins - pharmacology
Humans
Keloid - drug therapy
Keloid - genetics
Keloid - metabolism
Male
Original
Real-Time Polymerase Chain Reaction
RNA, Messenger - genetics
RNA, Small Interfering - genetics
Transfection
Up-Regulation
의학일반
Heat shock proteins
keloids
Hsp70
Online AccessGet full text

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Abstract There are currently no consistently effective treatments for the excessive collagen produced by keloid fibroblasts. Previously, we reported that heat shock protein 70 (Hsp70) is up-regulated in keloid fibroblasts and keloid tissue. We, therefore, investigated whether Hsp70 is related to excessive collagen production in keloid fibroblasts. We inhibited Hsp70 in keloid fibroblasts by RNA interference and examined the resulting collagen expression. Thus, we selected small interfering RNAs (siRNAs) specific for human Hsp70, transfected them into keloid fibroblasts, and evaluated the resulting phenotypes and protein production using real-time polymerase chain reaction (PCR), Western blot, and a collagen assay. The siRNAs dramatically suppressed Hsp70 mRNA expression, resulting in a decrease in collagen production in the keloid fibroblasts compared with controls. The siRNAs did not influence the viability of the keloid fibroblasts. Hsp70 overexpression likely plays an important role in the excessive collagen production by keloid fibroblasts. RNA interference has therapeutic potential for the treatment of keloids.
AbstractList PURPOSEThere are currently no consistently effective treatments for the excessive collagen produced by keloid fibroblasts. Previously, we reported that heat shock protein 70 (Hsp70) is up-regulated in keloid fibroblasts and keloid tissue. We, therefore, investigated whether Hsp70 is related to excessive collagen production in keloid fibroblasts.MATERIALS AND METHODSWe inhibited Hsp70 in keloid fibroblasts by RNA interference and examined the resulting collagen expression. Thus, we selected small interfering RNAs (siRNAs) specific for human Hsp70, transfected them into keloid fibroblasts, and evaluated the resulting phenotypes and protein production using real-time polymerase chain reaction (PCR), Western blot, and a collagen assay.RESULTSThe siRNAs dramatically suppressed Hsp70 mRNA expression, resulting in a decrease in collagen production in the keloid fibroblasts compared with controls. The siRNAs did not influence the viability of the keloid fibroblasts.CONCLUSIONHsp70 overexpression likely plays an important role in the excessive collagen production by keloid fibroblasts. RNA interference has therapeutic potential for the treatment of keloids.
Purpose: There are currently no consistently effective treatments for the excessive collagen produced by keloid fibroblasts. Previously,we reported that heat shock protein 70 (Hsp70) is up-regulated in keloid fibroblasts and keloid tissue. We, therefore, investigatedwhether Hsp70 is related to excessive collagen production in keloid fibroblasts. Materials and Methods: We inhibited Hsp70 in keloid fibroblasts by RNA interference and examined the resulting collagen expression. Thus, we selected small interfering RNAs (siRNAs) specific for human Hsp70, transfected them into keloid fibroblasts, and evaluated the resulting phenotypes and protein production using real-time polymerase chain reaction (PCR), Western blot, and a collagen assay. Results: The siRNAs dramatically suppressed Hsp70 mRNA expression, resulting in a decrease in collagen production in the keloidfibroblasts compared with controls. The siRNAs did not influence the viability of the keloid fibroblasts. Conclusion: Hsp70 overexpression likely plays an important role in the excessive collagen production by keloid fibroblasts. RNA interference has therapeutic potential for the treatment of keloids. KCI Citation Count: 12
There are currently no consistently effective treatments for the excessive collagen produced by keloid fibroblasts. Previously, we reported that heat shock protein 70 (Hsp70) is up-regulated in keloid fibroblasts and keloid tissue. We, therefore, investigated whether Hsp70 is related to excessive collagen production in keloid fibroblasts. We inhibited Hsp70 in keloid fibroblasts by RNA interference and examined the resulting collagen expression. Thus, we selected small interfering RNAs (siRNAs) specific for human Hsp70, transfected them into keloid fibroblasts, and evaluated the resulting phenotypes and protein production using real-time polymerase chain reaction (PCR), Western blot, and a collagen assay. The siRNAs dramatically suppressed Hsp70 mRNA expression, resulting in a decrease in collagen production in the keloid fibroblasts compared with controls. The siRNAs did not influence the viability of the keloid fibroblasts. Hsp70 overexpression likely plays an important role in the excessive collagen production by keloid fibroblasts. RNA interference has therapeutic potential for the treatment of keloids.
Author Shin, Jung U
Jung, Inhee
Tran, Thanh-Nga
Lee, Ju Hee
Lee, Won Jai
AuthorAffiliation 2 Department of Plastic and Reconstructive Surgery, Severance Hospital, Institute for Human Tissue Restoration, Yonsei University College of Medicine, Seoul, Korea
1 Department of Dermatology, Severance Hospital, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Korea
3 Department of Dermatology and Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
AuthorAffiliation_xml – name: 1 Department of Dermatology, Severance Hospital, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Korea
– name: 3 Department of Dermatology and Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
– name: 2 Department of Plastic and Reconstructive Surgery, Severance Hospital, Institute for Human Tissue Restoration, Yonsei University College of Medicine, Seoul, Korea
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  givenname: Thanh-Nga
  surname: Tran
  fullname: Tran, Thanh-Nga
  organization: Department of Dermatology and Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
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  surname: Jung
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  organization: Department of Dermatology, Severance Hospital, Cutaneous Biology Research Institute, Yonsei University College of Medicine, Seoul, Korea. juhee@yuhs.ac
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Issue 6
Keywords Heat shock proteins
keloids
Hsp70
Language English
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Snippet There are currently no consistently effective treatments for the excessive collagen produced by keloid fibroblasts. Previously, we reported that heat shock...
PURPOSEThere are currently no consistently effective treatments for the excessive collagen produced by keloid fibroblasts. Previously, we reported that heat...
Purpose: There are currently no consistently effective treatments for the excessive collagen produced by keloid fibroblasts. Previously,we reported that heat...
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StartPage 1619
SubjectTerms Adolescent
Adult
Blotting, Western
Collagen - drug effects
Collagen - metabolism
Female
Fibroblasts - metabolism
Gene Expression Regulation
HSP70 Heat-Shock Proteins - genetics
HSP70 Heat-Shock Proteins - metabolism
HSP70 Heat-Shock Proteins - pharmacology
Humans
Keloid - drug therapy
Keloid - genetics
Keloid - metabolism
Male
Original
Real-Time Polymerase Chain Reaction
RNA, Messenger - genetics
RNA, Small Interfering - genetics
Transfection
Up-Regulation
의학일반
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Title Hsp70 Knockdown by siRNA Decreased Collagen Production in Keloid Fibroblasts
URI https://www.ncbi.nlm.nih.gov/pubmed/26446645
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