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Tumor Biology |
and HIF-1 Target Genes during Multi-Stage Epidermal Carcinogenesis and Wound Healing1
Cutaneous Oncology Program, University of California-San Francisco Comprehensive Cancer Center [D. A. E., J. W. S., J. M. A.], and Department of Surgery, University of California-San Francisco School of Medicine [J. M. A.], San Francisco, California 94143-1674, and the Department of Biology, University of California-San Diego [H. E. R., R. J.], San Diego, California 92093-0366
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ABSTRACT |
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 Top ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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, and three targets of the heterodimeric transcription factor
HIF-1, glucose transporter (GLUT)-1, phosphoglycerate kinase (PGK)-1,
and vascular endothelial growth factor (VEGF) in skin. Neither
HIF-1, GLUT-1, PGK-1, nor VEGF mRNA was detectable in unwounded
nontransgenic skin. In epidermal carcinogenesis, HIF-1, GLUT-1,
PGK-1, and VEGF mRNAs were just detectable in early-stage hyperplasia,
markedly increased in high-grade epidermal chest dysplasias, and
further increased in invasive squamous carcinomas. In neoplastic skin,
HIF-1, GLUT-1, and PGK-1 mRNAs localized in the basal and immediate
suprabasal epidermal layers, whereas VEGF mRNA was predominantly
expressed in the more superior spinous and granular epidermal layers.
Immediately after wounding, HIF-1,
GLUT-1, and PGK-1 mRNAs were detectable in basal
keratinocytes at the wound edge. Expression of all three genes
increased to maximum levels in reepithelializing basal keratinocytes
and then diminished to near undetectable levels after wound
epithelialization. Although VEGF mRNA similarly increased and decreased
during wound healing, its expression pattern was more punctate; the
most intense hybridization signals were detected in the upper spinous
and granular layers of reepithelializing keratinocytes and in dermal
cells morphologically similar to macrophages. These data suggest
stage-specific and spatio-temporal control of HIF-1 and HIF-1 target
gene expression in both multistage epithelial carcinogenesis and wound
healing. |
INTRODUCTION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Regions of hypoxia are common in both healing wounds and growing masses
of malignant tissue (4)
. In both cancers and healing
wounds, hypoxia alters overall cellular behavior, as a consequence of,
or in addition to, activating specific genetic pathways. In cancers,
hypoxic regions are resistant to chemo- or radiation therapy, yet also
harbor zones of enhanced apoptosis (4)
. In wounds, low
oxygen appears to modulate the pace of healing and wound resolution and
specifically to increase collagen synthesis and accelerate wound
reepithelialization (5)
. This latter effect may be
attributable to enhancement of keratinocyte motility and migration by
hypoxia (6)
. At the level of gene expression and
transcriptional activation, hypoxia alters the activity of selected
transcription factors including
HIF3
-1, c-jun via ref-1 and/or c-jun NH2-terminal
kinase, and p53 (7, 8, 9, 10, 11)
. HIF-1 is a heterodimeric
transcription factor composed of HIF-1 and HIF-1ß
(12)
. HIF-1 protein is ubiquitinated and degraded in
normoxia (13
, 14)
but stabilized in hypoxia, whereas
HIF-1ß or aryl hydrocarbon nuclear translocator protein is
constitutively expressed in both normoxia and hypoxia. HIF-1
transactivates a repertoire of genes facilitating metabolic and
vascular adaptation to hypoxia (15)
. These genes include
several glycolytic enzymes, GLUT1 and 3, VEGF, inducible nitric oxide
synthase, and erythropoietin. Gene knockouts of either HIF-1 component
demonstrate embryonic lethality at days 9–11 attributable to
inhibition of vasculogenesis, mesenchymal cell death, and cardiac
defects (16, 17, 18)
. Several studies document hypoxic
induction of HIF-1 protein and up-regulation of HIF-1 target genes
in established cancer cell lines. Immunohistochemical analysis
demonstrates the presence of HIF-1 protein in variety of human cancers
and in selected premalignant pathologies such as benign prostatic
hyperplasia and colonic adenoma (19
, 20)
. However, to
date, no study has systematically investigated HIF-1 expression at
each stage of carcinogenic progression. Similarly, despite evidence of
hypoxia, increased angiogenesis, and VEGF up-regulation, there are no
data regarding HIF-1 expression during wound healing.
K14-HPV16 transgenic mice develop spontaneous multistage epidermal
carcinogenesis beginning with hyperplasia at 1–2 months of age,
dysplasia at 4 months of age, followed by premalignant
hyperproliferative follicular/interfollicular chest dysplasias or
dysplastic sessile papillomas at 8 months of age, each of which lead to
invasive squamous cancers (21)
. Previously, we
demonstrated VEGF mRNA induction in keratinocytes, particularly in
late-stage premalignant precursors in K14-HPV16 transgenic mice
(22)
. Here, we show induction of HIF-1 mRNA at a
threshold level in hyperplastic skin of transgenic mice, which is then
markedly abundant in premalignant hyperproliferative
follicular/interfollicular chest dysplasias (21)
and
further increased in cancers. Three targets of the HIF-1 transcription
factor, GLUT-1, PGK-1, and VEGF are coordinately up-regulated at
similar stages of premalignant neoplastic progression. However,
although the expression pattern of GLUT-1 and PGK-1 parallel that of
HIF-1 mRNA in neoplastic basal keratinocytes, VEGF mRNA is expressed
in more superior upper spinous and granular epidermal cell layers.
HIF-1, GLUT-1, and PGK-1 mRNAs are also up-regulated in epidermal
keratinocytes in a spatio-temporal expression pattern during wound
healing. These genes are undetectable in nontransgenic skin, are
expressed at high levels in basal keratinocytes that are
reepithelializing the open wound, and then decrease to nearly
undetectable levels when the wound is completely epithelialized and
dermal scar formation is under way. Similar to carcinogenesis, the
expression pattern of VEGF mRNA in reepithelializing keratinocytes
appeared to be different compared with HIF-1, GLUT-1, and PGK-1.
Collectively, these data suggest coordinate gain of HIF-1 expression
and HIF-1 function in proliferating basal squamous epithelial cells at
specific stages of carcinogenesis and wound healing. Moreover, this
study suggests that regulation of HIF-1 gene expression
in cancer progression and wound healing may occur at the level of mRNA
in addition to that of protein (8
, 23)
.
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MATERIALS AND METHODS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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and HIF-1 target gene expression during multi-stage
carcinogenesis was determined using K14-HPV16 transgenic mice of the
1203#1 line backcrossed (n = 28) into the
FVB/n inbred strain (21
, 24) . These mice develop
spontaneous, invasive, epidermal squamous carcinomas predominantly from
8 to 12 months of age, most frequently in the ear or on the chest and
lower neck (21
, 25)
. Wound healing experiments were
performed using 8–12-week-old nontransgenic male and female FVB/n
mice. All procedures and protocols were reviewed and approved by the
University of California Committee on Animal Research.
Wound Healing.
Mice were anesthetized with 2.5% Avertin, and their backs were shaved.
Incisions 
8 mm long were made through the skin, leaving the
underlying fasciae intact, and spread apart to prevent healing by
primary closure. Mice were housed individually and serially sacrificed
under Avertin anesthesia on days 1, 3, 5, 7, 9, and 11 after wounding
by bilateral thoracotomy, followed by intraventricular perfusion with
ice-cold PBS and 3.75% paraformaldehyde. An ellipse of skin 1
cm2 surrounding the wound was removed and
postfixed overnight in 3.75% paraformaldehyde at 4°C. Wounds were
bisected perpendicular to the axis of the incision and embedded in
paraffin.
Histopathology.
Routine histopathology was performed using H&E staining of 5-µm
sections.
Tissue DNA Synthesis.
BrdUrd (100 mg/kg) was injected i.p., 2 h prior to sacrifice, and
immunoperoxidase staining to detect BrdUrd incorporation into DNA was
performed on paraffin-embedded tissue sections as described previously
(21
, 24)
.
In Situ Hybridization.
Duplicate tissue sections from three different mice at each stage
of carcinogenesis or wounding were hybridized to
35S-labeled riboprobes as described previously
(26)
. The 356-bp HIF1 riboprobe DNA template
corresponded to nucleotides 513–869 of the mouse gene and was cloned
into pGEM-T easy (Pharmacia, Madison, WI). An antisense riboprobe was
generated by template linearization with SacII and in
vitro transcription with SP6 polymerase. The GLUT-1 riboprobe
template was 520 bp corresponding to nucleotides 275–794 of the mouse
gene cloned into pBSSK (Stratagene, La Jolla, CA). An antisense
riboprobe was generated by template linearization with PstI
and T3 polymerase transcription. The PGK-1 riboprobe template probe was
485 bp, from 91 to 575 of the mouse gene, cloned into pGEM-T easy. An
antisense riboprobe was generated by template linearization with
PstI and transcription with T7 polymerase. An antisense VEGF
riboprobe was generated as described previously (22)
.
Sense riboprobes were generated from each template and used as negative
controls. mRNA hybridization signals were determined from a qualitative
estimate of silver grain density that was reproducible in duplicate
sections from three different mice.
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RESULTS |
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mRNA and HIF-1 Target Genes during
Multistage Epidermal Carcinogenesis. and the HIF-1 target
gene GLUT-1 during multistage carcinogenesis, we used
35S mRNA in situ hybridization to
tissue sections from control nontransgenic and transgenic mice with
intermediate (16 weeks of age) and late-stage (32 weeks of age)
precursor lesions and invasive malignancies (Fig. 1)
. Precursor lesions from the 16-week-old transgenic mice were
predominately hyperplasias and intermediate-grade dysplasias, whereas
those of the 32-week-old transgenic mice were hyperproliferative
follicular/interfollicular dysplasias (Ref. 21
; Fig. 1
,
top row). Chest malignancies were moderately to poorly
differentiated squamous carcinomas (Fig. 1
, top row),
whereas ear cancers were well-differentiated sessile papillary
malignancies (data not shown). Neither HIF-1 nor GLUT-1 mRNA was
detectable in control, nontransgenic skin (Fig. 1
, left
panels). Hyperplastic chest skin displays a 2-fold increase in all
epidermal cell layers and a moderate infiltration of the dermis with
mononuclear cells (Fig. 1)
. Both HIF-1 and GLUT-1 mRNAs were just
detectable in the basal and immediate suprabasal epidermal cell layers
in these lesions (Fig. 1)
. Hyperproliferative
follicular/interfollicular chest dysplasias consist of marked
enlargement of the hair follicle ORS, suprabasal extension of
basal-type keratinocytes, and replacement of the uppermost stratum
corneum with nucleated parakeratotic cells (Fig. 1
; Ref.
21
). HIF-1 and GLUT-1 mRNAs were markedly increased in
these lesions (Fig. 1)
. Moreover, nearly all of the hybridization
signals for both mRNAs were localized to the basal layers of the
dysplastic interfollicular epidermis and the hair follicle ORS (Fig. 1)
. Hyperproliferative follicular/interfollicular chest dysplasias
immediately adjacent to chronic superficial wounds displayed the
highest level of signal for both HIF-1 and GLUT-1 mRNA (data not
shown). No hybridization of either HIF-1 or GLUT-1 mRNA was detected
in the dermis (Fig. 1)
. In moderate-poorly differentiated chest cancers
(Fig. 1
, right panels), hybridization signals for HIF-1
and GLUT-1 mRNAs further increased compared with hyperproliferative
follicular/interfollicular chest dysplasias, and was similarly
localized to epithelial squamous carcinoma cells compared with stromal
cells (Fig. 1)
. Both HIF-1 and GLUT-1 mRNA levels were increased in
dysplastic ear papillomas and papillary ear cancers; however,
hybridization signals were much fewer compared with the chest lesions,
suggesting that the carcinogenesis at this site was associated with a
lower induction of these mRNAs (data not shown).
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mRNA (Fig. 2)
. Both VEGF and HIF-1 mRNAs were initially, and barely, detectable
in the hyperplastic stage of carcinogenesis (Fig. 1
and Ref.
22
). Similarly, the hybridization signals for both VEGF
and HIF-1 mRNAs were further increased in premalignant
hyperproliferative follicular/interfollicular chest dysplasias (Ref.
21
; Fig. 2
). However, the distribution of VEGF and
HIF-1 mRNAs was markedly different in these precursor lesions.
HIF-1 expression was localized to the basal and immediate suprabasal
epidermal layers, whereas VEGF mRNA was predominantly detectable in the
upper spinous and lower granular epidermal layers (Fig. 2, A and B)
. In cancers, both HIF-1 and VEGF hybridization
signals further increased compared with hyperproliferative
follicular/interfollicular chest dysplasias; however, the expression
patterns of the mRNAs appeared to have subtle differences. Although
both HIF-1 and VEGF mRNAs were diffusely present throughout the
squamous epithelial component of the cancers, VEGF expression was
punctate and markedly increased (Fig. 2, C and D)
in what, on higher magnification, were individual malignant squamous
cells (data not shown).
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. Epidermal thickness at the wound edge further increased by day 3
(data not shown). Depending on the age of the mouse, by days 5–7, the
wound was nearly covered by a sheet of reepithelializing epidermal
cells migrating inward from both edges (Fig. 3)
. This epidermal sheet
was six to eight cell layers thick, whereas epidermis at a distance
from the wound edge was four cell layers in thickness (data not shown).
By days 11–13 after wounding, the epidermis remained thickened;
however, contracture markedly decreased the overall wound area with
epidermis of normal thickness drawn closer to the wound edge (Fig. 3)
.
Fibroblasts, destined to create the wound scar, were also prominent in
the underlying dermis (Fig. 3)
.
|
. On days 5–7, BrdUrd incorporation was seen in the basal
keratinocytes of the reepithelializing epidermal sheet and in scattered
cells in the dermis (Fig. 3)
. Elevation in BrdUrd incorporation was
restricted to the wound area and was similar to normal skin only
200–300 µm from the wound edge at all stages of wound healing (data
not shown). At resolution of wound reepithelialization by days 11–13,
BrdUrd incorporation was similar to unwounded skin, with only an
occasional positive basal keratinocyte (Fig. 3)
.
Spatio-Temporal Changes in HIF-1, GLUT-1, and PGK-1 mRNAs during
Wound Healing.
On day 1 after wounding, HIF-1 mRNA was barely detectable in
epidermal keratinocytes at the wound edge (Fig. 4)
. By day 5, HIF-1 message was abundant in the sheet of keratinocytes
reepithelializing the wound, in the same basal cell population, in
adjacent sections, as was incorporating BrdUrd (compare Figs. 3
and 4
).
By day 11 after wounding, HIF-1 message was undetectable in
epidermis (Fig. 4)
. At no time during wound healing was HIF-1 mRNA
detectable in dermis. Similar to DNA synthesis (see previous section),
HIF-1 expression also fell off precipitously such that no message
was detectable 200–300 µm from the wound edge (data not shown).
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subunit during each stage of wound healing (Fig. 4)
. GLUT-1
and PGK-1 mRNAs were detectable in basal and immediate suprabasal
keratinocytes on day 1 after skin incision at the wound edge. During
wound reepithelialization, there was a marked increase in GLUT-1 and
PGK-1 message, again predominantly localized to the basal and immediate
suprabasal layers. By day 11, GLUT-1 and PGK-1 mRNAs were barely
detectable in occasional basal keratinocytes. Similar to HIF-1,
GLUT-1 and PGK-1 mRNAs were not detectable in the dermis at any stage
of wound healing.
Comparison of HIF-1 and VEGF expression patterns during wound
healing also revealed similarities and differences, although the
differences were less striking than in carcinomas (Fig. 2)
. During
wound reepithelialization, VEGF mRNA displayed both a basal and a
suprabasal/granular cell layer component in the reepithelializing sheet
of keratinocytes at day 5 after wounding (Fig. 2F)
. Similar
to carcinomas, the VEGF expression pattern was punctate with marked
increases in VEGF mRNA in individual keratinocytes (Fig. 2F)
. In contrast, HIF-1 expression was diffusely
distributed throughout the basal and immediate suprabasal layers of the
reepithelializing keratinocyte sheet (Fig. 2E)
.
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DISCUSSION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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mRNA during epidermal carcinogenesis in K14-HPV16 transgenic mice and
wound healing in nontransgenic mice. Similar to a previous report
(20)
, HIF-1 mRNA is induced in the proliferative
compartment of the epidermis in both healing wounds and during
carcinogenesis. The proximity of these epidermal basal cells to the
underlying dermal capillaries (22)
suggests that this
compartment may not necessarily be hypoxic, and that increases in
HIF-1 mRNA may be induced by signals activating proliferation, such
as growth factors. Our data suggest that spatio-temporal gain of HIF-1
function may contribute to malignant conversion of late-stage
premalignant epithelial dysplasias and to wound reepithelialization.
Previous reports suggest that the predominant regulation of HIF-1
expression is via inhibition of protein ubiquitination and breakdown by
hypoxia (13)
. Ubiquitination of HIF-1 protein is
coordinated by a 200-amino acid "oxygen dependent degradation
domain" (13)
. Both von Hippel-Lindau protein and p53
have been shown to bind HIF-1 and may contribute to normoxic protein
instability by recruiting ubiquitin ligases such as CUL-1 or mdm-2,
respectively (27
, 28)
. p53 may also inhibit
HIF-1-mediated target gene transactivation (29)
.
Modulation of HIF-1 function by p53 is a link to HPV, because the E6
oncoprotein of high-risk viruses, such as type 16, bind to p53 with
high affinity and catalyze its ubiquitination and destruction
(30)
. As such, HPV16 E6 expression has been shown to
increase HIF-1 protein expression, DNA binding, transcriptional
activity, and inhibit HIF-1 ubiquitination and destruction
(28)
. In contrast, other work demonstrates that hypoxia
inhibits E6/E6-associated protein-mediated p53 destruction, suggesting
that augmentation of HIF-1 protein levels in hypoxic
HPV16-expressing cells may occur by mechanisms independent of loss of
p53 function (11)
. A major difference between these
experiments is that the level of hypoxia was greater in the Alarcon
et al. (0.02% O2; Ref.
11
) compared with the Ravi et al. (1.0%
O2; Ref. 28
) study, with the
possibility that an additional "stress" response coordinated by
c-jun, c-jun NH2-terminal kinase, or ref-1, may
have been induced by more profound hypoxia (10
, 31 , 32)
.
Regardless of these discrepancies, increases in HPV E6
oncogene expression may explain the up-regulation of HIF-1 target genes
in late-stage precursor lesions in K14-HPV16 transgenic mice.
Previously, we demonstrated increased basal keratinocyte E6
and E7 transgene expression during multistage epidermal
carcinogenesis in this model (25)
. Thus, incremental E6
expression would be consistent with the marked up-regulation of GLUT-1
and PGK-1 in late-stage premalignant precursor lesions and in invasive
epidermal cancers by stabilization of the HIF-1 component of HIF-1.
HIF-1 and HPV16 may also cooperate at the level of cellular
metabolism. Alterations in the activity and oligomerization of the m2
isoform of pyruvate kinase (m2PK) by the HPV16 E7 oncoprotein is a
potential link between enhanced aerobic glycolysis, HIF-1 function,
and HPV carcinogenesis. Binding of HPV16 E7 to m2PK favors formation of
m2PK dimers rather than tetramers (33)
. Dimerization of
m2PK decreases its affinity for pyruvate, shifting enzyme activity
toward lactate production via aerobic glycolysis (33)
.
Although pyruvate kinase is not a direct HIF-1 target gene,
transactivation of lactate dehydrogenase would further facilitate
lactate production and aerobic glycolysis in cells expressing HPV E7.
As such, coordinate functions of both HPV E7 and HIF-1/HIF-1 could
enhance the metabolic adaptation of HPV expressing neoplastic or
malignant cells to focal regions of hypoxia within growing cancers.
There have been few reports focusing on HIF-1 expression at the
level of mRNA. In most instances, HIF-1 mRNA levels are found to be
similar in transformed cells compared with nontransformed cells.
However, retroviral oncogenes, v-src in particular, have
been shown to increase HIF-1 mRNA in cells after transfection
(34)
. Our study clearly demonstrates incremental increases
in epidermal basal cell mRNA during both epidermal carcinogenesis and
wound healing. Although this increase in mRNA may occur by both
transcriptional as well as posttranscriptional mechanisms, examination
of the HIF-1 enhancer-promoter DNA sequence provides a
provocative link to epidermal carcinogenesis and HPV disease in
particular. There are at least two Sp-1 sites 5' to the start site and
an AP-1 site in the 5' untranslated region (35
, 36)
. The
presence of the AP-1 site may be significant. Enhanced AP-1 expression
and transcriptional activity has been demonstrated in multi-stage skin
carcinogenesis in mice transgenic with bovine papillomaviral oncogenes
(37)
. Moreover, loss of c-fos function (38)
or dominant-negative inhibition of c-jun (39)
each prevent
epidermal carcinogenesis induced by chemical carcinogens, further
highlighting the role of AP-1-mediated signaling in skin
carcinogenesis. As such, it will be of interest to determine whether
loss or inhibition of AP-1 function in these models is also associated
with less HIF-1 expression.
In addition to hypoxia, another level of HIF-1 regulation is growth
factor signaling. Insulin-like growth factor-1, insulin-like growth
factor-2, epidermal growth factor, and basic fibroblast growth factor
have all been shown to increase HIF-1 expression
(40, 41, 42)
. Moreover, growth factor and hypoxic control of
HIF-1 protein stability appear to be independent and parallel
pathways (41)
. We have demonstrated constitutive basic
fibroblast growth factor expression and up-regulation of acidic
fibroblast growth factor in the basal cell layer during epidermal
carcinogenesis in K14-HPV16 transgenic mice (26)
.
Moreover, we have unpublished data showing marked elevation of basal
keratinocyte transforming growth factor- expression in the
dysplastic stage and in invasive epidermal cancers in this model. In
our current study, up-regulation of HIF-1 mRNA in the basal cell
epidermal cell layer in both multi-stage carcinogenesis and wound
healing was correlated with increased proliferation during both
neoplastic progression in K14-HPV16 transgenic mice (24
, 25)
and wound healing in nontransgenic mice. Additionally, the
highest level of HIF-1 mRNA hybridization signals were in
premalignant precursor lesions and cancers adjacent or in the midst of
chronic superficial ulcers, which would be expected to contain both
stromal and epidermal cells with increased wound-associated growth
factor expression (1)
. Thus, our work, similar to previous
reports, (20)
appears to link HIF-1 expression, and
possibly HIF-1 transcriptional activity, to epithelial cell
proliferation and elevated growth factor expression.
Several studies, including our own, highlight angiogenesis, and in
particular VEGF up-regulation, as frequent and important components of
both multistage epidermal carcinogenesis and wound healing (22
, 43
, 44
, 45)
. In the current study, the stage specificity of
up-regulation of HIF-1 and VEGF mRNAs in K14-HPV16 transgenic mice
during carcinogenesis and wound healing were similar, although the
distribution and patterns of expression of the respective mRNAs were
different. Because HIF-1 is a nuclear transcription factor, detection
of VEGF and HIF-1 mRNAs in both similar and distinct epidermal cell
layers is consistent with both HIF-1-dependent and -independent
pathways of regulation of VEGF expression during carcinogenesis and
wound healing. In particular, macrophage VEGF expression
(46)
during wound healing is likely to be HIF-1
independent, because we did not detect macrophage HIF-1 mRNA at any
stage of wound healing.
In summary, we have demonstrated increased basal epidermal keratinocyte
HIF-1 mRNA expression in the skin of K14-HPV16 transgenic mice
during multi-stage carcinogenesis and during wound healing in
nontransgenic mice. Coordinate increase of HIF-1 target gene expression
in the same cell population, and at similar stages of either
carcinogenesis or wound healing, is consistent with increased HIF-1
function with the caveat that independent and parallel mechanisms for
up-regulation of these target genes may also exist. Ultimately,
engineering targeted gain or loss of keratinocyte HIF-1 function
will establish the necessity and role of this molecule in epithelial
carcinogenesis and wound healing.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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1 Supported by Grant NCI R01-CA71398 from the
National Cancer Institute. 
2 To whom requests for reprints should be
addressed, at Department of Surgery, University of California-San
Francisco Comprehensive Cancer Center, 2340 Sutter Street, Box 1674,
San Francisco, CA 94143-1674; Phone: (415) 885-3617; Fax:
(415) 885-7617; E-mail: jarbeit{at}cc.uscf.edu
3 The abbreviations used are: HIF, hypoxia
inducible factor; GLUT, glucose transporter; PGK, phosphoglycerate
kinase; VEGF, vascular endothelial growth factor; CUL, cullen; HPV,
human papillomavirus; ORS, outer root sheath of hair follicle; K14,
keratin 14; BrdUrd, 5-bromo-2-deoxyuridine.
Received 4/ 1/00. Accepted 8/29/00.
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REFERENCES |
|---|
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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subunit. J. Biol. Chem., 271: 32253-32259, 1996. is mediated by an O2-dependent degradation domain via the ubiquitin-proteasome pathway. Proc. Natl. Acad. Sci. USA, 95: 7987-7992, 1998. by the ubiquitin-proteasome pathway. J. Biol. Chem., 274: 6519-6525, 1999.-null embryos is not associated with VEGF deficiency but with mesenchymal cell death. Dev. Biol., 209: 254-267, 1999.[Medline]
. Genes Dev., 12: 149-162, 1998. is required for solid tumor formation and embryonic vascularization. EMBO J., 17: 3005-3015, 1998.[Medline]
in rat and human prostate cancer. Cancer Res., 58: 5280-5284, 1998. in common human cancers and their metastases. Cancer Res., 59: 5830-5835, 1999.. Genes Dev., 14: 34-44, 2000. locus. Gene Exp., 6: 287-299, 1997.
and insulin-like growth factor 2. Cancer Res., 59: 3915-3918, 1999.This article has been cited by other articles:
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|
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|
 |
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|
|
|
|
 |
|
 G. Fitsialos, I. Bourget, S. Augier, A. Ginouves, R. Rezzonico, T. Odorisio, F. Cianfarani, T. Virolle, J. Pouyssegur, G. Meneguzzi, et al. HIF1 transcription factor regulates laminin-332 expression and keratinocyte migration J. Cell Sci., September 15, 2008; 121(18): 2992 - 3001. [Abstract] [Full Text] [PDF]
|
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|
 |
|
 S. Roy, S. Khanna, C. Rink, S. Biswas, and C. K. Sen Characterization of the acute temporal changes in excisional murine cutaneous wound inflammation by screening of the wound-edge transcriptome Physiol Genomics, July 1, 2008; 34(2): 162 - 184. [Abstract] [Full Text] [PDF]
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 |
|
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|
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|
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|
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 H. Tanaka, M. Yamamoto, N. Hashimoto, M. Miyakoshi, S. Tamakawa, M. Yoshie, Y. Tokusashi, K. Yokoyama, Y. Yaginuma, and K. Ogawa Hypoxia-Independent Overexpression of Hypoxia-Inducible Factor 1{alpha} as an Early Change in Mouse Hepatocarcinogenesis Cancer Res., December 1, 2006; 66(23): 11263 - 11270. [Abstract] [Full Text] [PDF]
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 Q. Ke and M. Costa Hypoxia-Inducible Factor-1 (HIF-1) Mol. Pharmacol., November 1, 2006; 70(5): 1469 - 1480. [Abstract] [Full Text] [PDF]
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E. Gottfried, L. A. Kunz-Schughart, S. Ebner, W. Mueller-Klieser, S. Hoves, R. Andreesen, A. Mackensen, and M. Kreutz Tumor-derived lactic acid modulates dendritic cell activation and antigen expression Blood, March 1, 2006; 107(5): 2013 - 2021. [Abstract] [Full Text] [PDF]
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 S. M. O'Reilly, M. O. Leonard, N. Kieran, K. M. Comerford, E. Cummins, M. Pouliot, S. B. Lee, and C. T. Taylor Hypoxia induces epithelial amphiregulin gene expression in a CREB-dependent manner Am J Physiol Cell Physiol, February 1, 2006; 290(2): C592 - C600. [Abstract] [Full Text] [PDF]
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 Z. Yun, Q. Lin, and A. J. Giaccia Adaptive Myogenesis under Hypoxia Mol. Cell. Biol., April 15, 2005; 25(8): 3040 - 3055. [Abstract] [Full Text] [PDF]
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 Y. W. Park, M. N. Younes, S. A. Jasser, O. G. Yigitbasi, G. Zhou, C. D. Bucana, B. N. Bekele, and J. N. Myers AEE788, a Dual Tyrosine Kinase Receptor Inhibitor, Induces Endothelial Cell Apoptosis in Human Cutaneous Squamous Cell Carcinoma Xenografts in Nude Mice Clin. Cancer Res., March 1, 2005; 11(5): 1963 - 1973. [Abstract] [Full Text] [PDF]
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 A. Hoeben, B. Landuyt, M. S. Highley, H. Wildiers, A. T. Van Oosterom, and E. A. De Bruijn Vascular Endothelial Growth Factor and Angiogenesis Pharmacol. Rev., December 1, 2004; 56(4): 549 - 580. [Abstract] [Full Text] [PDF]
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 O. Stoeltzing, M. F. McCarty, J. S. Wey, F. Fan, W. Liu, A. Belcheva, C. D. Bucana, G. L. Semenza, and L. M. Ellis Role of Hypoxia-Inducible Factor 1{alpha} in Gastric Cancer Cell Growth, Angiogenesis, and Vessel Maturation J Natl Cancer Inst, June 16, 2004; 96(12): 946 - 956. [Abstract] [Full Text] [PDF]
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 Y.-J. Jung, J. S. Isaacs, S. Lee, J. Trepel, and L. Neckers Microtubule Disruption Utilizes an NFkappa B-dependent Pathway to Stabilize HIF-1alpha Protein J. Biol. Chem., February 21, 2003; 278(9): 7445 - 7452. [Abstract] [Full Text] [PDF]
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T. Daikoku, H. Matsumoto, R. A. Gupta, S. K. Das, M. Gassmann, R. N. DuBois, and S. K. Dey Expression of Hypoxia-inducible Factors in the Peri-implantation Mouse Uterus Is Regulated in a Cell-specific and Ovarian Steroid Hormone-dependent Manner. EVIDENCE FOR DIFFERENTIAL FUNCTION OF HIFs DURING EARLY PREGNANCY J. Biol. Chem., February 21, 2003; 278(9): 7683 - 7691. [Abstract] [Full Text] [PDF]
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 D. Baatar, M. K. Jones, K. Tsugawa, R. Pai, W. S. Moon, G. Y. Koh, I. Kim, S. Kitano, and A. S. Tarnawski Esophageal Ulceration Triggers Expression of Hypoxia-Inducible Factor-1{alpha} and Activates Vascular Endothelial Growth Factor Gene : Implications for Angiogenesis and Ulcer Healing Am. J. Pathol., October 1, 2002; 161(4): 1449 - 1457. [Abstract] [Full Text] [PDF]
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 C. Pellizzaro, D. Coradini, and M. G. Daidone Modulation of angiogenesis-related proteins synthesis by sodium butyrate in colon cancer cell line HT29 Carcinogenesis, May 1, 2002; 23(5): 735 - 740. [Abstract] [Full Text] [PDF]
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 J.M. ARBEIT Quiescent Hypervascularity Mediated by Gain of HIF-1{alpha} Function Cold Spring Harb Symp Quant Biol, January 1, 2002; 67(0): 133 - 142. [Abstract] [PDF]
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J. E. Albina, B. Mastrofrancesco, J. A. Vessella, C. A. Louis, W. L. Henry Jr., and J. S. Reichner HIF-1 expression in healing wounds: HIF-1alpha induction in primary inflammatory cells by TNF-alpha Am J Physiol Cell Physiol, December 1, 2001; 281(6): C1971 - C1977. [Abstract] [Full Text] [PDF]
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 E. Schipani, H. E. Ryan, S. Didrickson, T. Kobayashi, M. Knight, and R. S. Johnson Hypoxia in cartilage: HIF-1alpha is essential for chondrocyte growth arrest and survival Genes & Dev., November 1, 2001; 15(21): 2865 - 2876. [Abstract] [Full Text] [PDF]
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 M. Crowther, N. J. Brown, E. T. Bishop, and C. E. Lewis Microenvironmental influence on macrophage regulation of angiogenesis in wounds and malignant tumors J. Leukoc. Biol., October 1, 2001; 70(4): 478 - 490. [Abstract] [Full Text] [PDF]
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D. A. Elson, G. Thurston, L. E. Huang, D. G. Ginzinger, D. M. McDonald, R. S. Johnson, and J. M. Arbeit Induction of hypervascularity without leakage or inflammation in transgenic mice overexpressing hypoxia-inducible factor-1{alpha} Genes & Dev., October 1, 2001; 15(19): 2520 - 2532. [Abstract] [Full Text] [PDF]
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