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J Biol Chem, Vol. 274, Issue 30, 20874-20878, July 23, 1999
From the Mesangial cells share features with contractile
smooth muscle cells and mechanically support the capillary wall. The
role of vitamin D compounds and the transforming growth factor- 1,25-Dihydroxyvitamin D3
(1,25(OH)2D3)1
was originally identified as a nutritional factor and a fat-soluble
vitamin. Its physiological role is to regulate calcium metabolism. It
was then reevaluated and came to be thought of as a kind of steroid
hormone. The role of 1,25(OH)2D3 has also been
recognized as being much broader and thus much more physiologically
important than originally thought, as reflected by its effects on the
proliferation and differentiation of a variety of cells mainly
including malignant and immune cells.
Since the first report (1) that 1,25(OH)2D3
inhibited proliferation and induced the differentiation of murine
myeloid leukemic M1 cells into monocyte macrophages, many studies have
demonstrated that 1,25(OH)2D3 is a potent
modulator in suppressing proliferation and inducing the differentiation
of numerous normal and tumor cells. However, the effective doses for
these effects often cause hypercalcemia. To prevent this, many
analogs of 1,25(OH)2D3 have been synthesized.
One of them, 22-oxa-1,25-dihydroxyvitamin D3 (22-oxacalcitriol, OCT), has a more potent differentiation-inducing effect (2) than 1,25(OH)2D3 without inducing hypercalcemia.
In the kidney, the anti-proliferative effects of
1,25(OH)2D3 have been demonstrated in cultured
mesangial cells (3). However, its differentiation-inducing effects have
remained unknown. Mesangial cells (MCs) are uniquely differentiated
cells, which acquire a smooth muscle-like phenotype, and are believed
to be derived from mesenchymal cells (4). We have previously reported
that cultured mesangial cells in the early passages (MC-E) (<25)
showed a smooth muscle cell phenotype such as low cell turnover, high
levels of expression for type IV collagen (Col IV) and smooth muscle
In addition, evidence is accumulating that suggests that MCs, during
the process of glomerular injuries (such as inflammation), can
proliferate and undergo a phenotypic modulation in which they markedly
up-regulate their expression of smooth muscle-like proteins (i.e. SMA). Furthermore, MCs can develop fibroblast-like
characteristics by secreting interstitial collagens (i.e.
type I and III collagens) that are not normally present in the
mesangial matrix (6, 7). However, there is little information regarding
the mechanisms that underlie this phenotypic change.
TGF- The loss of responsiveness to the inhibitory effects of TGF- Recent evidence indicates that a loss of RII is often associated
with a failure to respond to autocrine and exogenous TGF- Vitamin D3 Compounds--
1,25(OH)2
D3 was purchased from Philips Duphar Co. (Amsterdam, The
Netherlands), and its synthetic analog OCT was generously supplied by
Chugai Pharmaceutical Co. Ltd. (Tokyo, Japan). Stock solutions were
prepared in absolute ethanol at 10 Cell Culture--
A glomerular mesangial cell line was
established from glomeruli isolated from normal, 4-week-old mice
(C57BL/6JxSJL/J) and was identified according to the method previously
described (13, 14). The mesangial cells were plated on 100-mm plastic
dishes (Nunc) and maintained in B medium (a 3:1 mixture of minimal
essential medium/F12 modified with trace elements) supplemented with 1 mM glutamine, penicillin at 100 units/ml, streptomycin at
100 mg/ml, and 20% fetal calf serum (Irvine Scientific). The cells
were passaged weekly with trypsin-EDTA. The cultured cells fulfilled
the criteria generally accepted for glomerular MCs previously described
(4, 15).
Cell Proliferation--
[3H]Thymidine
incorporation was performed to assess the DNA synthesis rate. The
mesangial cells were plated at 2.5 × 104 cells/well in
24-well plates for 24 h, and the medium was replaced with
serum-free medium containing 0.1% bovine serum albumin and the
incubation continued for another 24 h. Various concentrations of
compounds (1, 25(OH)2D3, OCT, or vehicle) and
[3H]thymidine (1 m Ci/ml, 1 µCi/well, NEN Life Science
Products) were then added, and the cells were incubated for 24 h
in 2.0% fetal calf serum. The cells were washed, precipitated with
10% trichloroacetic acid, and counted in a liquid scintillation
counter (16). The experiments were performed in quadruplicate.
RNase Protection Assay--
Total RNA was isolated from
mesangial cells using the TRIzol reagent (Life Technologies, Inc.) and
was analyzed by a RNase protection assay with the following probes. The
RNA probes were prepared by linearizing the PvuII fragment
of Col IV ( TGF- Mink Lung Epithelial Cell Growth Inhibition Assay--
The cells
were plated onto 100-mm dishes and allowed to reach 70-80%
confluency. The medium was removed and replaced with 5 ml of fresh B
medium. The cells were then treated with
1,25(OH)2D3, OCT, or vehicle only for 24 h. Following this treatment, the conditioned medium was collected, and
the indicated volumes were used to treat mink lung epithelial cells
plated on 96-well plates at a density of 1500 cells/well. In addition,
a standard TGF- DNA Transfection--
The Col IV ( Statistical Analysis--
Data were expressed as mean ± S.E. and analyzed by an unpaired Student's t test.
Vitamin D3 Sensitivity--
The effects of
1,25-dihydroxyvitamin D3 and its analog on the cell
proliferation of MC-E and MC-L cells were investigated by assessing the
[3H]thymidine incorporation following treatment by these
compounds. Both 1,25(OH)2D3 and OCT inhibited
DNA incorporation of the MC-E cells in a dose-dependent
manner. The effect of OCT is more prominent than that of
1,25(OH)2D3 (Fig.
1A). In contrast, these
compounds did not significantly affect the MC-L cells (Fig.
1B).
Expression of SMA, Col IV, and Col I mRNA--
The mRNAs
for SMA, Col IV, and GAPDH were easily detectable in the untreated MC-E
cells (Fig. 2A), whereas Col I
mRNA was poorly detected. After exposure to
1,25(OH)2D3 or OCT, the OCT treatment showed an
approximately 1.5- and 4-fold induction of SMA and Col IV mRNA
respectively, whereas the expression levels of Col I remained
unchanged. 1,25(OH)2D3 treatment did not result in any change. In contrast, treatment with these compounds showed no
change in the SMA and Col IV mRNA expression of the MC-L cells (Fig. 2B); however, the Col I mRNA levels were increased
2.2-fold in the OCT-treated MC-L cells. The GAPDH mRNA levels were
equivalent in all cells.
Transcriptional Activity of SMA and Col IV--
Transcriptional
activity was investigated by transiently transfecting cells with fusion
constructs containing a promoter/enhancer/CAT reporter for each gene.
The OCT-treated cells or control cells were transfected with the target
constructs. The transcriptional activity of the SMA gene promoter was
analyzed by the construct SMP-1, which contained the SMA promoter, the
SV40 enhancer, and the CAT reporter gene. The CAT activity was
increased 3.7-fold in the OCT-treated cells as compared with the
control cells, demonstrating that OCT enhanced the transcriptional
activity of the SMA gene. A p56 construct was used to evaluate the
transcriptional activity of Col IV. OCT induced a 9.4-fold increase in
CAT activity as compared with the control, thus indicating that OCT
significantly stimulated the promoter activity of the Col IV ( TGF- Active TGF- Expression of RII mRNA Levels--
The other possibility for
the increased autocrine TGF- MCs possess a specific smooth muscle phenotype including
contractile properties, an extensive array of microfilaments, and the
production of basement components. In several studies, cultured MCs
exhibited an increasingly dedifferentiated phenotype as they were
passaged (27, 28). These dedifferentiated MCs can produce interstitial
collagen and gain the capability to proliferate. In this study, we
first demonstrated that a vitamin D3 analog could induce a
smooth muscle phenotype in cultured MCs. The induction of smooth muscle
phenotype could be related to TGF- The signal transduction of TGF- Thus, 1,25(OH)2D3 and its analog can inhibit
cell proliferation through the induction of negative autocrine TGF- A dedifferentiated smooth muscle phenotype can be observed in cultured
MCs after multiple passages. These dedifferentiated cells have
decreased RII expression and have lost their responsiveness to TGF- *
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
¶
To whom correspondence should be addressed: Division of
Artificial Kidneys, Faculty of Medicine, Kyoto University, 54 Shogoin-Kawaharacho, Sakyo-ku Kyoto 606-8507, Japan. Tel.:
81-75-751-3642; Fax: 81-75-751-3643; E-mail:
doitoshi@kuhp.kyoto-u.ac.jp.
The abbreviations used are:
1,25(OH)2D3, 1,25-dihydroxyvitamin
D3;
OCT, 22-oxa-1,25-dihydroxyvitamin D3;
TGF-
A Vitamin D Analog Regulates Mesangial Cell Smooth Muscle
Phenotypes in a Transforming Growth Factor-
Type II
Receptor-mediated Manner*
,,,, and Division of Molecular Medicine for Adult and
Geriatric Diseases,
ABSTRACT
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
(TGF-) type II receptor in modulating the smooth muscle phenotype of cultured mesangial cells was examined. Cell proliferation was significantly inhibited by the vitamin D analog
22-oxa-1,25-dihydroxyvitamin D3 (22-oxacalcitriol;
OCT) rather than by 1,25-dihydroxyvitamin D3
(1,25(OH)2D3) in a dose-dependent
manner. OCT-treated early passage mesangial cells (MC-E cells) had
increased expression levels of type IV collagen and smooth muscle 
actin mRNA, but 1,25(OH)2D3-treated MC-E
cells did not. The addition of a TGF-1-neutralizing antibody to the OCT-treated MC-E cells blocked this inhibitory effect
for cell proliferation and attenuated the up-regulated mRNA levels.
However, after exposure to 1,25(OH)2D3 or OCT,
there was no significant difference in the secretion of active TGF-. We next investigated whether TGF- type II receptor (RII) was involved in this regulation. OCT treatment significantly increased the
expression of the RII mRNA in MC-E cells. These results suggest that the vitamin D analog OCT induces smooth muscle phenotypic alterations and that this phenomenon was mediated through the induction
of RII in cultured mesangial cells.
INTRODUCTION
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ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
actin (SMA), together with low expression of type I collagen (Col
I). After multiple passages, mesangial cells in the late passages (MC-L) (>45) lost their smooth muscle phenotype and increased cell
turnover (5).
has widespread effects on extracellular matrix proliferation in
many cultured cell lines and appears to play a role in the pathological
accumulation of the extracellular matrix that accompanies inflammatory
and fibrotic diseases such as glomerulonephritis (8). TGF- may also
have other important functions on glomerular epithelial and mesangial
cells, including the regulation of cell proliferation, hypertrophy, and
survival (apoptosis), as well as the modulation of the local and
systemic immune response. Recent studies have shown that TGF-
overexpression in experimental and human kidney disease leads to
progressive glomerular and tubulointerstitial scarring and renal
failure (9-11).
has
been implicated in the development of a variety of cancers. The signal
transduction of TGF- is regulated via a heteromeric complex of type
I and type II TGF- receptors (RI and RII). An attenuation of TGF-
receptor expression is likely to play a critical role in the escape
from this growth regulation.
. In some
cancer cell lines, RII expression is certainly a critical determinant
for conferring TGF- tumor suppression as well as negative autocrine
TGF- growth functions. A study by Wu et al. (12) in human
breast cancer cells demonstrated that increased RII mRNA and
protein levels following treatment with
1,25(OH)2D3 and its analogs contributed to
TGF- activity. It is generally accepted that
1,25(OH)2D3 works in context with TGF- in
controlling cell proliferation. However, with respect to cell
differentiation, it is still undetermined how
1,25(OH)2D3 and TGF- interact. In this
study, we investigated the mechanisms underlying the differentiation of
the MC phenotype in vitro.
EXPERIMENTAL PROCEDURES
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ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
4 M. Serial
dilutions were then made in absolute ethanol and stored at 
20 °C
protected from light. These diluted solutions were added to the
experimental culture medium at a final ethanol concentration of 1%.
The control cells received 1% ethanol vehicle, which had no effect on
cell proliferation (data not shown).
1) from p1234, the HindIII fragment of SMA
from pSA2HE, the ApaI fragment of Col I (2) from pGM101,
and the EcoRI fragment of GAPDH from pMGAP1 (17-19). In
addition, we constructed a RII riboprobe to measure the RII mRNA
levels. A fragment of mouse RII cDNA (196 base pairs) was obtained
by polymerase chain reaction using the primer pair
5'-GCAAGTTTTGCGATGTGAGA-3' and 5'-GCATCTTCCAGAGTGAAGCC-3', according to
the sequence published by Suzuki et al. (20). The polymerase
chain reaction fragment was confirmed to be the RII cDNA by DNA
sequencing and was cloned into a pCR II-TOPO plasmid (Invitrogen).
After digesting the plasmid with SacI, an antisense riboprobe was synthesized in vitro using T7 RNA polymerase,
which protects a 196-base pair fragment. The RNA probe (4 × 105 cpm) and the test RNA were hybridized overnight at
45 °C. RNase A (40 µg/ml) and RNase T1 (2 µg/ml) were added to
each tube, and the tubes were incubated for 1 h at 30 °C. The
RNase resistant fragments were analyzed by 3.5 or 7% polyacrylamide-8
M urea gel electrophoresis and autoradiography (5, 21). The
protected bands for each RNA probe had the same size as the coding
sequence for the specific mRNA, thus providing evidence for their
specificity, and were evaluated by densitometric analysis.
-neutralizing Antibody Assay--
The cells were
resuspended at a concentration of 5 × 104 cells/ml and
plated onto 24-well plates (500 µl/well) in the presence of either 10 µg/ml TGF-1 neutralizing antibody (R&D Systems) or a control
normal IgG. After 3 h of incubation different concentrations of
vitamin D3 compounds were added as indicated. The cells
were allowed to grow for 72 h without changing the medium followed by the determination of the [3H]thymidine incorporation
as described above.
growth inhibition curve was generated by treating
the cells with various concentrations of TGF-1. The mink lung
epithelial cells were allowed to incubate for 3 days, at which time the
medium was removed and replaced by 100 µl of fresh medium. The
colonies were immediately visualized by staining with
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (Sigma)
for 2 h. The stained cells were solubilized with Me2SO
(Sigma), and the relative cell numbers were determined by the resultant
absorbance at 595 nm.
1)
promoter/enhancer/chloramphenicol acetyltransferase (CAT) construct
(p56) has been previously described (22). The construct SMP-1 contains
1074 base pairs of the proximal 5'-flanking region plus 43 base pairs
of the 5'-untranslated region of the SMA gene cloned into the
SalI-BamHI site of the SV40 enhancer containing
CAT vector pBLCAT3 (23). Next, 4 × 105 cells were plated
on a 60-mm dish and transfected with plasmid DNA using the
LipofectAMINE PLUS Reagent (Life Technologies, Inc.). For each
transfection, 2 µg of CAT reporter plasmid and 0.5 µg of
pSV--galactosidase (Promega) (internal control for transfection efficiency) were used. Forty eight hours after the start of the transfection the cells were harvested and assayed for CAT and -galactosidase activity (24, 25). CAT (Amersham Pharmacia Biotech)
or -galactosidase (Promega) was used as the standard, and pSV0CAT
and pSV2CAT were used for negative (0%) and positive (100%) controls,
respectively (26). A correction factor for differences in the
transfection efficiency was obtained by normalizing the CAT activity to
the -galactosidase activity and then subtracting the background
level, as determined by transfection with pSV0CAT. The final value was
expressed as the relative CAT activity.
RESULTS
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ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
View larger version (23K):
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Fig. 1.
Effects of
1,25(OH)2D3 and OCT on DNA synthesis in MC-E
(A) and MC-L (B) cells. The
cells were plated onto 24-well plates at a density of 2.5 × 104 cells/well. After synchronization in serum-free medium
containing 2.0% fetal calf serum supplemented with
1,25(OH)2D3 and OCT at a final concentration of
10 10 to 106 M or vehicle (1%
ethanol). The [3H]thymidine incorporation into DNA was
measured and expressed as a percentage of control untreated with
vitamin D3 compounds. The data represent the mean ± S.E. of four determinations.
View larger version (65K):
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Fig. 2.
Effects of
1,25(OH)2D3 and OCT on the expression of
specific mRNAs in MC-E and MC-L cells. After treatment with
10 10 to 106 M
1,25(OH)2D3 (lanes 2, 3,
and 4), OCT (lanes 5, 6, and
7), or vehicle alone (lane 1) for 24 h in
the MC-E (A) and MC-L cells (B), the specific
mRNAs were determined by using RNase protection assay. The amount
of total RNA loaded (1 µg/lane) was checked by hybridization with a
GAPDH probe. The data are representative of three independent
experiments. CTL, control; VD,
1,25(OH)2D3.
1)
gene (Fig. 3). These data suggest that
increased mRNA levels for SMA and Col IV (1) induced by OCT were
because of increased promoter activity in their genes.
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Fig. 3.
Effects of OCT on transcriptional
activity. The transient transfection followed by treatment with
10 6 M OCT was performed on the MC-E cells,
and CAT activity was determined 48 h after transfection. The
values are presented as -fold increase calculated with respect to that
obtained in the absence of the OCT (vehicle only), which was defined as
1. The data shown are the mean ± S.E. of triplicate
determinations.
Autocrine Activity--
To elucidate the correlation
between TGF- and vitamin D3 sensitivity, a
TGF--neutralizing antibody assay was performed. At 10 µg/ml, the
TGF--neutralizing antibody reversed the inhibitory effects of
vitamin D3 and its analog OCT; the antibody generated an
approximately 50% increase in DNA synthesis as compared with normal
chicken IgG treatment, in a dose-dependent manner, on MC-E cells (Fig. 4A). In contrast,
the MC-L cells did not respond to the TGF--neutralizing antibody
(Fig. 4B), thus indicating a lack of induction of autocrine
TGF- activity. These results indicate that the proliferation
inhibitory mechanisms of vitamin D3 involve the induction
of TGF- activity in MC-E cells. Moreover, the addition of
TGF--neutralizing antibody attenuated the up-regulated mRNA levels (data not shown).
View larger version (18K):
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Fig. 4.
Effects of a
TGF- -neutralizing antibody on DNA synthesis in
1,25(OH)2D3 or OCT-treated mesangial
cells. The MC-E (A) and MC-L cells (B) were
plated at a density of 2.5 × 104 cells/well. After the
addition of the TGF--neutralizing antibodies and vitamin
D3 compounds, [3H]thymidine incorporations
were measured. The results were represented as a percentage increase in
DNA synthesis relative to their respective control antibody-treated
cells. The data shown are the mean ± S.E. of triplicate
determinations.
Levels in MCs Treated with Conditioned
Medium--
The increased autocrine TGF- activity could have
resulted from the enhanced expression of TGF- and/or its receptors.
To test these possibilities, an inhibition bioassay on mink lung epithelial cells was performed. As a standard, the addition of TGF-1
inhibited the proliferation of MC-E cells in a
dose-dependent fashion (data not shown). Conditioned medium
from 1,25(OH)2D3 (106
M) or OCT (106 M)-treated and
-untreated MC-E cells was added to mink lung epithelial cells. After
exposure to either treated or untreated conditioned medium, no
significant differences in proliferation inhibition were observed in
the mink lung epithelial cells (Fig. 5).
Similarly, no significant differences in proliferation inhibitory
effects were observed in the MC-L cells (data not shown). Moreover, the other conditioned medium, 1010 or 108
M 1,25(OH)2D3 or OCT, resulted in
no significant difference. These results demonstrated that vitamin
D3 compounds did not alter the activation of secreted
growth and/or inhibitory cytokines from MC-E cells, one of which is
likely to be TGF-1. Furthermore, these results also suggest that the
enhanced TGF- autocrine activity by the vitamin D3
compounds did not result from the modulation of ligand activation.
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Fig. 5.
Determination of the levels of active
TGF- in conditioned medium from
1,25(OH)2D3 or OCT-treated and -untreated MC-E
cells. After treatment with 106 M
1,25(OH)2D3 (
), OCT (×), or vehicle alone
(
) in the MC-E cells, conditioned media were collected. Various
volumes indicated were added to mink lung epithelial cells to perform
proliferation inhibition assay. The data are represented as a
percentage inhibition in cell proliferation relative to each value in
the absence of conditioned medium.
activity following treatment with
vitamin D3 compounds was the induction of receptor
expression; therefore, we determined whether
1,25(OH)2D3 and its analog could modulate the
expression of RII mRNA. 1,25(OH)2D3 (106 M) or OCT (106
M) were utilized to determine the kinetic effects on RII
expression. MC-E cells untreated with vitamin D3 compounds
expressed RII mRNA 6-fold higher than MC-L cells. After exposure to
1,25(OH)2D3 and OCT, a 2.1- and 4.5-fold
increase, respectively, in RII mRNA expression levels in MC-E cells
was observed. On the other hand, OCT-treated MC-L cells showed a
1.8-fold increase in their RII mRNA levels, but no significant
modulation was noted in 1,25(OH)2D3-treated MC-L cells (Fig. 6).
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Fig. 6.
Regulation of TGF-
RII mRNA expression by 1,25(OH)2D3
and OCT. After treatment with 106 M
1,25(OH)2D3 (lanes 2 and
5), OCT (lanes 3 and 6), and vehicle
alone (lanes 1 and 4) in the MC-E (lanes
1-3) and MC-L cells (lanes 4-6) for 24 h, the
specific mRNAs were determined by using the RNase protection assay.
The amount of total RNA loaded (1 µg/lane) was checked by
hybridization with a GAPDH probe. The data are representative of two
independent experiments.
DISCUSSION
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
signaling. However, active
TGF- levels were not enhanced by the vitamin D3
compounds in this study. The increased cellular responsiveness to
TGF- was correlated with a significant induction of RII expression by the vitamin D3 analog. This study suggests that the
vitamin D3 analog may be a new modulator for the smooth
muscle phenotype and that the mechanisms underlying this process are
tightly related to the regulation of RII expression.
is mediated via two different types
of serine/threonine kinase receptors, RI and RII. RII is the primary
binding protein for ligands and is the most important for signal
transduction. RII transactivates RI, which transduces various signals
via the Smad family (29-31). Next, we examined the levels of RII
mRNA expression. The low expression of RII mRNA in the MC-L
cells supports the connection that there was no inhibition of cell
proliferation because of the vitamin D3 compounds.
Increased RII mRNA expression was observed after treatment with OCT
rather than 1,25(OH)2D3 in the MC-E cells,
corresponding to the higher ability of OCT to inhibit cell
proliferation. The TGF--neutralizing antibody prevented inhibitory
effects of cell proliferation by vitamin D3 compounds.
These effects seemed to be correlated with the increasing amounts in
RII expression induced by 1,25(OH)2D3 or OCT.
Therefore, these data suggest that TGF- signal transduction significantly prevents these effects of vitamin D3
compounds. Moreover, the stimulatory and inhibitory Smad families may
alter the feedback after OCT treatment.
activity. However, the effective dose of
1,25(OH)2D3, which could induce negative
autocrine TGF- activity would also cause hypercalcemia, thus leading
to unwanted adverse effects. To overcome this problem, 1,25(OH)2D3 analogs such as OCT have been
developed that have increased potency and a reduced hypercalcemic
effect (32). We have demonstrated here that the analog OCT has similar
effects as 1,25(OH)2D3 at lower concentrations,
which make it an attractive and potential anti-proliferative agent
against proliferative glomerulonephritis. The marked induction of
differentiation caused by OCT was similar to the phenomena observed in
leukemia cells, T cells, breast cancer cells, fibroblasts, and
keratinocytes (2, 33-35). To elucidate the mechanisms responsible for
the progression of glomerular injuries, it is indispensable to
appreciate the phenotypic changes as well as the proliferation of
mesangial cells. TGF- has been implicated in the differentiation of
vascular smooth muscle cells and other mesenchyme-derived cells during
development (36). For example, the TGF- treatment of human smooth
muscle cells (37), granulation tissue myofibroblasts (38), and
pericytes (39) has been shown to increase the expression of SMA, a
marker of differentiated smooth muscle cells (40). However, the
molecular basis for this phenotypic modulation and differentiation in
cultured MCs is as yet poorly understood. Furthermore, OCT may offer an
advantage in that the induction of the autocrine negative TGF-
activity occurs through RII and not the TGF- ligand and thus could
avoid damaging adjacent tissues.
,
which may be related to the dedifferentiation process. On the other
hand, RII can be detected on MCs in vivo according to our
recent observations. These findings suggest that RII could be a key
regulator of MC differentiation and that OCT is an important modulator
for RII expression. Further work is in progress to clarify the role of
RII in the differentiation mechanisms of MCs in vivo and
in vitro.
FOOTNOTES
ABBREVIATIONS
, transforming growth factor-;
RI, TGF- type I receptor;
RII, TGF- type II receptor;
SMA, smooth muscle actin;
Col IV, type IV collagen;
Col I, type I collagen;
GAPDH, glyceraldehyde-3-phosphate dehydrogenase;
MC, mesangial cell;
MC-E/L, early/late passage mesangial cells;
CAT, chloramphenicol
acetyltransferase.
REFERENCES
TOP
ABSTRACT
INTRODUCTION
EXPERIMENTAL PROCEDURES
RESULTS
DISCUSSION
REFERENCES
1.
Abe, E.,
Miyaura, C.,
Sakagami, H.,
Takeda, M.,
Konno, K.,
Yamazaki, T.,
Yoshiki, S.,
and Suda, T.
(1981)
Proc. Natl. Acad. Sci. U. S. A.
78,
4990-4994 2.
Abe, J.,
Morikawa, M.,
Miyamoto, K.,
Kaiho, S.,
Fukushima, M.,
Miyaura, C.,
Abe, E.,
Suda, T.,
and Nishii, Y.
(1987)
FEBS Lett.
226,
58-62[CrossRef][Medline]
[Order article via Infotrieve]
3.
Weinreich, T.,
Merke, J.,
Schonermark, M.,
Reichel, H.,
Diebold, M.,
Hansch, G. M.,
and Ritz, E.
(1991)
Am. J. Kidney Dis.
18,
359-366[Medline]
[Order article via Infotrieve]
4.
Striker, G. E.,
and Striker, L. J.
(1985)
Lab. Invest.
53,
122-131[Medline]
[Order article via Infotrieve]
5.
Iehara, N.,
Takeoka, H.,
Tsuji, H.,
Imabayashi, T.,
Foster, D. N.,
Strauch, A. R.,
Yamada, Y.,
Kita, T.,
and Doi, T.
(1996)
Kidney Int.
49,
1330-1341[Medline]
[Order article via Infotrieve]
6.
Floege, J.,
Johnson, R. J.,
Gordon, K.,
Iida, H.,
Pritzl, P.,
Yoshimura, A.,
Campbell, C.,
Alpers, C. E.,
and Couser, W. G.
(1991)
Kidney Int.
40,
477-488[Medline]
[Order article via Infotrieve]
7.
Johnson, R. J.,
Floege, J.,
Yoshimura, A.,
Iida, H.,
Couser, W. G.,
and Alpers, C. E.
(1992)
J. Am. Soc. Nephrol.
2 (suppl.),
190-197
8.
Okuda, S.,
Languino, L. R.,
Ruoslahti, E.,
and Border, W. A.
(1990)
J. Clin. Invest.
86,
453-462
9.
Yamamoto, T.,
Noble, N. A.,
Cohen, A. H.,
Nast, C. C.,
Hishida, A.,
Gold, L. I.,
and Border, W. A.
(1996)
Kidney Int.
49,
461-469[Medline]
[Order article via Infotrieve]
10.
Yamamoto, T.,
Nakamura, T.,
Noble, N. A.,
Ruoslahti, E.,
and Border, W. A.
(1993)
Proc. Natl. Acad. Sci. U. S. A.
90,
1814-1818 11.
Yamamoto, T.,
Noble, N. A.,
Miller, D. E.,
and Border, W. A.
(1994)
Kidney Int.
45,
916-927[Medline]
[Order article via Infotrieve]
12.
Wu, G.,
Fan, R. S.,
Li, W.,
Srinivas, V.,
and Brattain, M. G.
(1998)
J. Biol. Chem.
273,
7749-7756 13.
Doi, T.,
Vlassara, H.,
Kirstein, M.,
Yamada, Y.,
Striker, G. E.,
and Striker, L. J.
(1992)
Proc. Natl. Acad. Sci. U. S. A.
89,
2873-2877 14.
MacKay, K.,
Striker, L. J.,
Elliot, S.,
Pinkert, C. A.,
Brinster, R. L.,
and Striker, G. E.
(1988)
Kidney Int.
33,
677-684[Medline]
[Order article via Infotrieve]
15.
Davies, M.
(1994)
Kidney Int.
45,
320-327[Medline]
[Order article via Infotrieve]
16.
Doi, T.,
Striker, L. J.,
Elliot, S. J.,
Conti, F. G.,
and Striker, G. E.
(1989)
Am. J. Pathol.
134,
395-404[Abstract]
17.
Oberbaumer, I.,
Laurent, M.,
Schwarz, U.,
Sakurai, Y.,
Yamada, Y.,
Vogeli, G.,
Voss, T.,
Siebold, B.,
Glanville, R. W.,
and Kuhn, K.
(1985)
Eur. J. Biochem.
147,
217-224[Medline]
[Order article via Infotrieve]
18.
Schenborn, E. T.,
and Mierendorf, R. C., Jr.
(1985)
Nucleic Acids Res.
13,
6223-6236 19.
Melton, D. A.,
Krieg, P. A.,
Rebagliati, M. R.,
Maniatis, T.,
Zinn, K.,
and Green, M. R.
(1984)
Nucleic Acids Res.
12,
7035-7056 20.
Suzuki, A.,
Shioda, N.,
Maeda, T.,
Tada, M.,
and Ueno, N.
(1994)
FEBS Lett.
355,
19-22[CrossRef][Medline]
[Order article via Infotrieve]
21.
Doi, T.,
Striker, L. J.,
Kimata, K.,
Peten, E. P.,
Yamada, Y.,
and Striker, G. E.
(1991)
J. Exp. Med.
173,
1287-1290 22.
Burbelo, P. D.,
Bruggeman, L. A.,
Gabriel, G. C.,
Klotman, P. E.,
and Yamada, Y.
(1991)
J. Biol. Chem.
266,
22297-22302 23.
Luckow, B.,
and Schutz, G.
(1987)
Nucleic Acids Res.
15,
5490 24.
Neumann, J. R.,
Morency, C. A.,
and Russian, K. O.
(1987)
BioTechniques
5,
1044-1051
25.
Rosenthal, N.
(1987)
Methods Enzymol.
152,
704-720[Medline]
[Order article via Infotrieve]
26.
Gorman, C. M.,
Moffat, L. F.,
and Howard, B. H.
(1982)
Mol. Cell. Biol.
2,
1044-1051 27.
Venkatachalam, M. A.,
and Kreisberg, J. I.
(1985)
Am. J. Physiol.
249,
C48-C55 28.
Floege, J.,
Radeke, H. R.,
and Johnson, R. J.
(1994)
Kidney Int.
45,
360-368[Medline]
[Order article via Infotrieve]
29.
Lin, H. Y.,
Wang, X. F.,
Ng-Eaton, E.,
Weinberg, R. A.,
and Lodish, H. F.
(1992)
Cell
68,
775-785[CrossRef][Medline]
[Order article via Infotrieve]
30.
Franzen, P.,
ten Dijke, P.,
Ichijo, H.,
Yamashita, H.,
Schulz, P.,
Heldin, C. H.,
and Miyazono, K.
(1993)
Cell
75,
681-692[CrossRef][Medline]
[Order article via Infotrieve]
31.
Bassing, C. H.,
Yingling, J. M.,
Howe, D. J.,
Wang, T.,
He, W. W.,
Gustafson, M. L.,
Shah, P.,
Donahoe, P. K.,
and Wang, X. F.
(1994)
Science
263,
87-89 32.
Murayama, E.,
Miyamoto, K.,
Kubodera, N.,
Mori, T.,
and Matsunaga, I.
(1986)
Chem. Pharm. Bull. (Tokyo)
34,
4410-4413[Medline]
[Order article via Infotrieve]
33.
Abe, J.,
Nakamura, K.,
Takita, Y.,
Nakano, T.,
Irie, H.,
and Nishii, Y.
(1990)
J. Nutr. Sci. Vitaminol.
36,
21-31
34.
Abe, J.,
Nakano, T.,
Nishii, Y.,
Matsumoto, T.,
Ogata, E.,
and Ikeda, K.
(1991)
Endocrinology
129,
832-837 35.
Koizumi, H.,
Kaplan, A.,
Shimizu, T.,
and Ohkawara, A.
(1997)
J. Dermatol. Sci.
15,
207-213[CrossRef][Medline]
[Order article via Infotrieve]
36.
Zentella, A.,
and Massague, J.
(1992)
Proc. Natl. Acad. Sci. U. S. A.
89,
5176-5180 37.
Bjorkerud, S.
(1991)
Arterioscler. Thromb.
11,
892-902 38.
Desmouliere, A.,
Geinoz, A.,
Gabbiani, F.,
and Gabbiani, G.
(1993)
J. Cell Biol.
122,
103-111 39.
Verbeek, M. M.,
Otte-Holler, I.,
Wesseling, P.,
Ruiter, D. J.,
and de Waal, R. M.
(1994)
Am. J. Pathol.
144,
372-382[Abstract]
40.
Owens, G. K.
(1995)
Physiol. Rev.
75,
487-517
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