Use of green fluorescent protein-conjugated β-actin as a novel molecular marker for in vitro tumor cell chemotaxis assay

Louis Hodgson, W. Qiu, C. Dong, A. J. Henderson

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

To study the dynamics of actin cytoskeleton rearrangement in living cells, an eukaryotic expression vector expressing a β-actin-GFP fusion protein was generated. The expression construct when transfected into NIH3T3 fibroblast, A2058 human melanoma and 293T human embryonic kidney carcinoma cell lines expressed β-actin-GFP fusion protein, which colocalized with endogenous cellular actin as determined by histoimmunofluorescence staining. The β-actin-GFP was also observed to be reorganized in response to treatments with the chemoattractant type IV collagen. Cells extended pseudopodial protrusions and altered the morphology of their cortical structure in response to type IV collagen stimulation. More importantly, β-actin-GFP accumulated in areas undergoing these dynamic cytoskeleton changes, indicating that β-actin-GFP could participate in actin polymerization. Although ectopic expression of β-actin-GFP lead to minor side effects on cell proliferation, these studies suggest that this strategy provides an alternative to the invasive techniques currently used to study actin dynamics and permits real-time visualization of actin rearrangements in response to environmental cues.

Original languageEnglish (US)
Pages (from-to)1106-1114
Number of pages9
JournalBiotechnology Progress
Volume16
Issue number6
DOIs
StatePublished - 2000
Externally publishedYes

Fingerprint

chemotaxis
Chemotaxis
Green Fluorescent Proteins
green fluorescent protein
actin
Actins
genetic markers
assays
Neoplasms
Collagen Type IV
collagen
neoplasm cells
In Vitro Techniques
chemoattractants
Chemotactic Factors
Eukaryotic Cells
melanoma
cytoskeleton
microfilaments
Cytoskeleton

ASJC Scopus subject areas

  • Food Science
  • Biotechnology
  • Microbiology

Cite this

Use of green fluorescent protein-conjugated β-actin as a novel molecular marker for in vitro tumor cell chemotaxis assay. / Hodgson, Louis; Qiu, W.; Dong, C.; Henderson, A. J.

In: Biotechnology Progress, Vol. 16, No. 6, 2000, p. 1106-1114.

Research output: Contribution to journalArticle

@article{d4e19ffa8a0f46c696b18c9130691c2b,
title = "Use of green fluorescent protein-conjugated β-actin as a novel molecular marker for in vitro tumor cell chemotaxis assay",
abstract = "To study the dynamics of actin cytoskeleton rearrangement in living cells, an eukaryotic expression vector expressing a β-actin-GFP fusion protein was generated. The expression construct when transfected into NIH3T3 fibroblast, A2058 human melanoma and 293T human embryonic kidney carcinoma cell lines expressed β-actin-GFP fusion protein, which colocalized with endogenous cellular actin as determined by histoimmunofluorescence staining. The β-actin-GFP was also observed to be reorganized in response to treatments with the chemoattractant type IV collagen. Cells extended pseudopodial protrusions and altered the morphology of their cortical structure in response to type IV collagen stimulation. More importantly, β-actin-GFP accumulated in areas undergoing these dynamic cytoskeleton changes, indicating that β-actin-GFP could participate in actin polymerization. Although ectopic expression of β-actin-GFP lead to minor side effects on cell proliferation, these studies suggest that this strategy provides an alternative to the invasive techniques currently used to study actin dynamics and permits real-time visualization of actin rearrangements in response to environmental cues.",
author = "Louis Hodgson and W. Qiu and C. Dong and Henderson, {A. J.}",
year = "2000",
doi = "10.1021/bp000093o",
language = "English (US)",
volume = "16",
pages = "1106--1114",
journal = "Biotechnology Progress",
issn = "8756-7938",
publisher = "John Wiley and Sons Ltd",
number = "6",

}

TY - JOUR

T1 - Use of green fluorescent protein-conjugated β-actin as a novel molecular marker for in vitro tumor cell chemotaxis assay

AU - Hodgson, Louis

AU - Qiu, W.

AU - Dong, C.

AU - Henderson, A. J.

PY - 2000

Y1 - 2000

N2 - To study the dynamics of actin cytoskeleton rearrangement in living cells, an eukaryotic expression vector expressing a β-actin-GFP fusion protein was generated. The expression construct when transfected into NIH3T3 fibroblast, A2058 human melanoma and 293T human embryonic kidney carcinoma cell lines expressed β-actin-GFP fusion protein, which colocalized with endogenous cellular actin as determined by histoimmunofluorescence staining. The β-actin-GFP was also observed to be reorganized in response to treatments with the chemoattractant type IV collagen. Cells extended pseudopodial protrusions and altered the morphology of their cortical structure in response to type IV collagen stimulation. More importantly, β-actin-GFP accumulated in areas undergoing these dynamic cytoskeleton changes, indicating that β-actin-GFP could participate in actin polymerization. Although ectopic expression of β-actin-GFP lead to minor side effects on cell proliferation, these studies suggest that this strategy provides an alternative to the invasive techniques currently used to study actin dynamics and permits real-time visualization of actin rearrangements in response to environmental cues.

AB - To study the dynamics of actin cytoskeleton rearrangement in living cells, an eukaryotic expression vector expressing a β-actin-GFP fusion protein was generated. The expression construct when transfected into NIH3T3 fibroblast, A2058 human melanoma and 293T human embryonic kidney carcinoma cell lines expressed β-actin-GFP fusion protein, which colocalized with endogenous cellular actin as determined by histoimmunofluorescence staining. The β-actin-GFP was also observed to be reorganized in response to treatments with the chemoattractant type IV collagen. Cells extended pseudopodial protrusions and altered the morphology of their cortical structure in response to type IV collagen stimulation. More importantly, β-actin-GFP accumulated in areas undergoing these dynamic cytoskeleton changes, indicating that β-actin-GFP could participate in actin polymerization. Although ectopic expression of β-actin-GFP lead to minor side effects on cell proliferation, these studies suggest that this strategy provides an alternative to the invasive techniques currently used to study actin dynamics and permits real-time visualization of actin rearrangements in response to environmental cues.

UR - http://www.scopus.com/inward/record.url?scp=0034523232&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034523232&partnerID=8YFLogxK

U2 - 10.1021/bp000093o

DO - 10.1021/bp000093o

M3 - Article

C2 - 11101341

AN - SCOPUS:0034523232

VL - 16

SP - 1106

EP - 1114

JO - Biotechnology Progress

JF - Biotechnology Progress

SN - 8756-7938

IS - 6

ER -