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Characterizing the protein expression of S1P receptors in T-cell acute lymphoblastic leukemia (T-ALL) cell lines

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Characterizing the protein expression of S1P receptors in T-cell acute lymphoblastic leukemia (T-ALL) cell lines

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Content CHARACTERIZING THE PROTEIN EXPRESSION OF S1P RECEPTORS IN
T-CELL ACUTE LYMPHOBLASTIC LEUKEMIA (T-ALL) CELL LINES
by
Yue Zhang
A Thesis Presented to the
FACULTY OF THE GRADUATE SCHOOL
UNIVERSITY OF SOUTHERN CALIFORNIA
In Partial Fulfillment of the
Requirements for the Degree
MASTER OF SCIENCE
(CRANIOFACIAL MOLECULAR BIOLOGY)
December 2008
Copyright 2008 Yue Zhang
ii
Table of Contents
List of Figures iii
Abstract iv
Introduction 1
Specific Aim 8
Experimental Design 9
Cell lines 10
Materials and Methods 12
Results 15
Discussion 21
References 24
iii
List of Figures
Figure 1: An overview of S1P signaling 1
Figure 2: Expression of EDG-1 receptor (42kd) in T-ALL cell lines 16
Figure 3: Expression of EDG-5 receptor (39kd) in T-ALL cell lines 17
Figure 4: Expression of EDG-6 receptor (42kd) in T-ALL cell lines 18
Figure 5: Testing EDG-3 (42kd) antibody with negative 19
and positive controls
Figure 6:Testing EDG-8 (46kd) antibody with negative 20
and positive controls
iv
Abstract
The implication of S1P in cancer has attracted researchers from a wide spectrum of fields,
but the relationship between S1P and leukemia has not been well studied, with limited
data indicating a possible anti-apoptotic and pro-survival role of S1P that may lead to
drug resistance. The current thesis aims at a characterization of the expression profile of
S1P receptors in T-cell ALL leukemia using western blot analysis. It is hypothesized that
one or more S1P receptors are expressed in a panel of T-ALL cell lines so that
S1P-induced signals are transduced by these receptors. We expect that western blot
analysis can detect the expression of these receptors. This project will lay the groundwork
for the future research on S1P signaling pathways in T-ALL cell lines and may
potentially provide more target molecules for drug development against T-cell ALL.
1
Introduction
Sphingosine-1-phosphate (S1P) is a product generated by the activation of sphingosine
kinase, which phosphorylates sphingsoine as the substrate. As a bioactive lipid
molecule, S1P has been demonstrated to exert a variety of effects on cell proliferation,
differentiation, migration and apoptosis, etc (Taha et al. 2006). Primarily secreted by
platelets, S1P exists extensively in human serum and reaches micromolar concentrations
(Murata et al. 2000;Yang et al. 1999),. Other sources of S1P also include erythrocytes,
neutrophils and mononuclear cells (Yang et al. 1999).
Figure 1: An overview of S1P signaling. Adapted from (Alvarez et al. 2007).
2
Leukemia is a cancer of blood cells characterized by multiplication of immature blood
cells and the subsequent loss of function. Acute lymphoblastic leukemia (ALL) features
over-produced immature white blood cells crowding out normal cells in the bone
marrow and metastasizing to other organs. It is the most common type of leukemia in
childhood in the U.S with a peak incidence at 4-5 years of age. Although nearly 90% of
low- and average-risk pediatric ALL can be cured, relapse has been a major challenge
due to drug resistance.
The mechanism of rapid multiplication of ALL cells has been a great interest of
researchers. In recent years S1P has been studied in various types of leukemia regarding
its pro-proliferative and cytoprotective effects. For example, in acute myeloid leukemia
(AML) cell line HL-60 S1P has been shown to elicit calcium influx(Okajima et al. 1996),
mediate cell differentiation(Sato et al. 1998), and antagonize apoptosis by inhibiting
cytochrome c release(Cuvillier and Levade 2001). In chronic myeloid leukemia (CML),
S1P mediates Abl-induced upregulation of Mcl-1(Li et al. 2007) and regulates resistance
to imatinib-induced apoptosis(Baran et al. 2007). Although it has been speculated that
S1P may exert similar effects of pro-proliferation and pro-survival in ALL cells, no
study has been reported regarding these effects and the corresponding mechanisms.
3
Extracellularly, S1P signals through a family of G-protein coupled receptors (GPCR),
known as Endothelial Differentiation Genes (EDG). Among the eight members of the
EDG family, five are S1P receptors, EDG1/S1P1, EDG5/S1P2, EDG3/S1P3,
EDG6/S1P4, EDG8/S1P5, and three lysophosphatidic acid (LPA) receptors,
EDG2/LPA1, EDG4/LPA2, EDG7/LPA3.
EDG1 has been demonstrated to play important roles in regulating cell
proliferation/survival, migration and cytoskeletal organization. Various cell lines, such
as human umbilical vascular endothelial cells (HUVEC), vascular smooth muscle cell
(VSMC) and Chinese hamster ovary (CHO) cells have shown pro-proliferative and
migratory responses to extracellular S1P (Hobson et al. 2001;Kon et al. 1999;Okamoto
et al. 1998;Okamoto et al. 2000;Zondag et al. 1998). Antisense knockdown of EDG1
impaired the survival and migration of HUVEC as well as its morphogenetic
differentiation (Lee et al. 1999;Paik et al. 2001), and EDG1 is also indispensable for
S1P-induced cortical actin and VE-cadherin localization at the intercellular junctions (
Lee et al. 1999). Other lines of research have demonstrated that S1P promotes
endothelial cell barrier integrity (Garcia et al. 2001) and increases membrane ruffling
(Okamoto et al. 2000). Recent research has also shown that EDG1 is important in
regulating the trafficking patterns of lymphocyte populations, such as controlling egress
4
of B and T cells from secondary lymphoid organs (Lo et al. 2005;Matloubian et al.
2004).
The signaling pathways mediated by EDG1 are dependent on its coupling to the
heterotrimeric G-protein, G
i
(Ancellin and Hla 1999;Windh et al. 1999) and small
GTPases such as rho and rac (Lee et al. 1998;Paik et al. 2001). For example, ERK1/2
activation mediated by EDG1 has been shown to be G
i
–dependent in various cell lines
(Lee et al. 1996;Okamoto et al. 1999). In addition, it has also been observed that
cytosolic calcium increases induced by S1P are regulated in part by EDG1 via G
i
(An et
al. 1999;Okamoto et al. 1999;Van, Jr. et al. 1998).
Although expressed as widely as EDG1, EDG5 seems to functionally differ from EDG1.
CHO cells forced to overexpress EDG5 do not exhibit enhanced migration in
comparison to those overexpressed with EDG1 (Kon et al. 1999). Another report
showed that EDG5 inhibited IGF-induced chemotaxis due to a proposed mechanism of
impairing the tivation of rac and p65 PAK (Okamoto et al. 2000). In terms of cell
proliferation, there has been research showing either pro-proliferative or
anti-proliferative effects of EDG5 in different systems (An et al. 2000;Van, Jr. et al.
1999). In mouse vascular endothelial cells, EDG5
5
is demonstrated to negatively regulate endothelial morphogenesis and
angiogenesis(Inoki et al. 2006).
In contrast to EDG1 that couples to only G
i ,
EDG5 mediates its signaling pathways
through G
i
, G
q
, G
13
(An et al. 1999;Ancellin and Hla 1999;Windh et al. 1999) as well
as small GTPase rho (Gonda et al. 1999). Various heterotrimeric G-proteins that EDG5
couples to might explain the activation of some signaling pathways such as
PLC-IP3-Ca
2+
(An et al. 1999;Gonda et al. 1999;Kupperman et al. 2000) and adenylyl
cyclase (Gonda et al. 1999;Kon et al. 1999),. MAP-kinases (ERK, JNK and p38) are
another group of signaling molecules downstream of EDG5. Although activation of
these kinases have been observed in different cell lines (An et al. 2000;Gonda et al.
1999), how JNK and p38, which may exhibit both pro-proliferative and pro-apoptotic
effects, interact with ERK remains unclear.
EDG3 shares similarities with EDG1 in terms of its wide expression in various tissues as
well as its cellular functions. For example, EDG3 promotes migration in CHO cells and
HUVEC (Kon et al. 1999;Okamoto et al. 2000), regulates cytoskeletal reorganization
and morphogenetic differentiation (Lee et al. 1999). But proliferation appears to be a
function that is not uniquely dependent on EDG3, since its overexpression stimulates
6
cell proliferation in hepatoma cells (An et al. 2000), while down-regulation of the
receptor in HUVEC does not impact S1P-induced cell survival (Lee et al. 1999).
Being able to couple to multiple heterotrimeric G-proteins, EDG3 can efficiently
activate PLC-IP3-Ca
2+
signaling pathways (An et al. 1999;Ishii et al. 2001;Sato et al.
1999). Regulation by EDG3 of adenylyl cyclase and MAP-kinases, such as ERK1/2 and
JNK, have also been widely studied, although reports on adenylyl cyclase showed
conflicting results (An et al. 1997;An et al. 2000;Kupperman et al. 2000);. Small
GTPase rac has also been shown to be activated in CHO cells and HUVEC (Okamoto et
al. 2000) (Paik et al. 2001), but its activation by EDG3 in mouse embryonic fibroblasts
is more controversial, as two studies have yielded conflicting results (Ishii et al.
2001;Liu et al. 2000).
EDG6 has a more restricted expression in the tissues and cells of hematopoietic systems
(Graler et al. 1998;Ishii et al. 2001). Research of recent years has revealed the cellular
functions of the receptor, including promoting cell migration when over-expressed in
CHO cells through activation of Cdc42 in a pertussis toxin-dependent manner (Kohno et
al. 2003) and stimulating T cell proliferation and cytokine secretion without signaling
migration (Wang et al. 2005). By coupling to G
i
and G
12/13
heterotrimeric G-proteins,
7
EDG6 also regulates cell shape and motility in CHO cells (Graler et al. 2003). Other
signaling pathways mediated by S1P4 include MAPK (ERK1/2) (Van, Jr. et al. 2000)
and PLC-Ca pathways(Yamazaki et al. 2000).
As the most recent member of the EDG family, EDG8 has not been well understood due
to lack of sufficient data. The mRNA expression of S1P5 in rats seems to be limited to
brain tissues, together with some conflicting reports on its expression in spleen (Ishii et
al. 2001;Yamazaki et al. 2000). In humans, the mRNA expression of EDG8 is wider,
including brain, heart and leukocytes, etc. (Niedernberg et al. 2002). Its cellular function
is relatively unique, too, due to its negative effect on cell proliferation, which is pertussis
toxin insensitive but orthovanadate sensitive (Yamazaki et al.2000). The heterotrimeric
G-proteins EDG8 couples to are G
i
and G
12
(Malek et al. 2001;Yamazaki et al. 2000)
and the downstream signaling effects include inhibiting adenylyl cyclase, activating JNK
while suppress ERK1/2 (Malek et al. 2001).
8
Specific Aim: Characterize the expression profile of S1P receptors in four T-ALL
cell lines.
We hypothesize that one or more S1P receptors are expressed at the protein level in
T-ALL cells. In particular, EDG-6 is likely to be expressed, since previous work has
demonstrated its predominant expression in hematopoietic systems and its effects on
T-cell trafficking and proliferation. (Graler et al. 1998a;Ishii et al. 2001e). Knowing the
expression profile of a number of T-ALL cells will allow future work to identify the
molecular pathways through which S1P can alter cell behavior.
9
Experimental Design
Western blot analysis will be used to determine the expression of individual S1P
receptors (EDG-1, EDG-3, EDG-5, EDG-6 and EDG-8). Both positive and negative
controls will serve as assistance for a more accurate detection. Human prostate protein
Medley (pools of total proteins isolated from human prostate) will be chosen as the
positive control for all of the five receptors, since western blotting data have shown that
all S1P receptors are expressed in the protein Medley(Duong et al. 2004). A gastric
cancer cell line, KATO III will be the negative control for EDG-1, EDG-3, EDG-6 and
EDG-8, and HL-60 (a chronic myeloid leukemia cell line) for EDG-5, because these two
cell lines do not express the corresponding receptors at the mRNA level (Sato et al.
1998b;Yamashita et al. 2006).
10
Cell Lines
Four T-ALL cell lines will be used for the experiments. MOLT-4 was established from
19-year-old male patient in relapse and has been used as a classic T-ALL cell line
(Minowada, et al., 1972). COG-LL-317, COG-LL-329 and COG-LL-332 were recently
established at Children's Hospital Los Angeles in Dr. Reynolds' laboratory.
COG-LL-317 was established from a 2-year-old male patient in the second relapse and
COG-LL-332 was established at relapse from the same patient (ten-year-old male) from
which COG-LL-329 was established at diagnosis (Sheard et al., 2007).
MOLT-4 is obtained from American Type Culture Collection (ATCC, Manassas, VA)
and cultured in RPMI-1640 (Mediatech, Herndon, VA) supplemented with 10%
heat-inactivated fetal bovine serum (FBS). COG-LL-317, COG-LL-329 and
COG-LL-332 are cultured in IMDM (Lonza, Walkersville, MD) supplemented with
20% FBS, 3mM L-Glutamine and 5ug/mL ITS (Insulin-transferrin-sodium selenite)
(Lonza, Walkersville, MD). All the cells are incubated in 37
o
C incubator with 5% O
2
,
mimicking the oxygen concentration in bone marrow.
For cell lines serving as negative controls, KATO III and HL-60 are both obtained from
ATCC (Manassas, VA). KATO III is cultured in IMDM media supplemented with 20%
11
FBS, as recommended by ATCC, in 37
o
C incubator with 20% O
2,
and HL-60 is cultured
RPMI-1640 media supplemented with 10% FBS in 37
o
C incubator with 5% O
2.
12
Materials and Methods
The cells are incubated in fresh media for 48 hours. One million cells are lysed
(Matsushita, et al., 2004) by lysis buffer (20 mM Tris pH 7.5, 150 mM NaCl, 1%
Nonidet P-40, 0.5% Sodium Deoxycholate, 1 mM EDTA and 0.1% SDS) and set on ice
for 10 min. Protease inhibitor cocktail (PMSF 1 mM, aprotinin 0.15 units/ml, Leupeptin,
10 uM, Pepstatin, 1 ug/ml and sodium fluoride 1 mM) is freshly added to the lysis
buffer. The cell lysates are centrifuged at 15,000 х g at 4
o
C for 20 min. A standard curve
is generated by diluting 1 mg/mL bovine serum albumin (BSA) to 0.5 mg/mL, 0.25
mg.mL, 0.125 mg/mL, and the protein concentration is measured using BCA reagents.
Cell lysates containing 30ug of protein are loaded onto a 15-well 4-12% tris-glycine gel,
which then runs in running buffer (25 mM Tris, 192 mM glycine and 0.1% SDS) at a
voltage of 120 V for 90 min.
The protein samples in the gel are transferred to PVDF membranes by electrophoresis
(100 V, 60 min, 4
0
C) in transfer buffer (25 mM Tris and 192 mM glycine) containing
20% methanol.
13
The membrane is blocked overnight at 4
0
C for non-specific binding in blocking buffer
which is 5% milk dissolved in TBS buffer (100 mM Tris and 150 mM NaCl) containing
0.1% Tween 20.
The membrane is incubated with primary antibody diluted 1:1000 at room temperature
for one hour, and washed three times in TBS buffer under constant motion, with each
wash lasting for 15 min. The antibodies against EDG-1, EDG-5, EDG-6 and EDG-8
are rabbit polyclonal antibodies and those against EDG-3 and beta-actin are goat
polyclonal antibodies
The membrane is then incubated with secondary antibody (HRP conjugated goat
anti-rabbit, or donkey anti-goat antibody) at room temperature for 1 hour and washed for
three times in TBS buffer, each wash lasts 15 min.
Femto supersensitive chemiluminescent substrate is applied to the membrane and the
film is developed in the dark room. Images are then documented by a scanner.
IMDM media (Lonza, Cat#:12-726F)
RPMI-1640 (Mediatech, Cat#: 10-040-CV)
ITS (Lonza, Cat#: 17-838-Z)
Lysis buffer (Invitrogen, Cat#: FNN0021)
14
Protease inhibitor (Sigma, Cat#: P8340)
BCA reagents (Pierce, Cat#: 23223 & 23224)
Tris-glycine gel (Invitrogen, Cat#:EC6508BOX)
Running buffer (Bio-rad, Cat#: 161-0072)
Transfer buffer (Bio-rad, Cat#: 161-0071)
Tween 20 (Sigma, 63158)
Femto Supersensitive Chemiluminescent (Pierce, Cat#:34080)
PVDF membrane (Rio-Rad, Cat#: 162-0176)
Autoradiography film(Denville, Cat#: E3012)
Human prostate protein Medley (Clontech, Cat#: 635336)
EDG-1 antibody (Santa Cruz, sc-25489)
EDG-3 antibody (Santa Cruz, sc-16076)
EDG-5 antibody (Santa Cruz, sc-25491)
EDG-6 antibody (Exalpha, x1631p)
EDG-8 antibody (Santa Cruz, sc-25493)
Beta actin antibody (Santa Cruz, sc-1615)
HRP conjugated goat anti-rabbit antibody (Santa Cruz, sc-3837)
HRP conjugated donkey anti-goat antibody (Santa Cruz, sc-3851)
15
Results
The expression of EDG-1(Figure 2), EDG-5 (Figure 3) and EDG-6 (Figure 4) has been
successfully detected by western blotting and confirmed by repetition. The experimental
conditions of the three antibodies are similar, regarding the blocking time (overnight,
4
0
C), primary antibody dilution (1:1000), secondary antibody dilution (1:10000) and
antibody incubation time (1 hour). The three S1P/EDG receptors are relatively evenly
expressed in T-ALL cell lines, MOLT-4, CHLA-317, CHLA-329 and CHLA-332.
16
Figure 2: Expression of EDG-1 receptor (42kd) in T-ALL cell lines
The membrane was blocked overnight at 4
0
C, and incubated with primary antibody
(1:1000 dilution) at room temperature for 1 hour. Secondary antibody was diluted
1:10000 and applied to the membrane for 1 hour.
17
Figure 3: Expression of EDG-5 receptor (39kd) in T-ALL cell lines
The membrane was blocked overnight at 4
0
C, and incubated with primary antibody
(1:1000 dilution) at room temperature for 1 hour. Secondary antibody was diluted
1:10000 and applied to the membrane for 1 hour.
18
Figure 4: Expression of EDG-6 receptor (42kd) in T-ALL cell lines
The membrane was blocked overnight at 4
0
C, and incubated with primary antibody
(1:1000 dilution) at room temperature for 1 hour. Secondary antibody was diluted
1:10000 and applied to the membrane for 1 hour.
19
EDG-3 antibody exhibited significantly reduced binding specificity, and the blot
obtained under aforementioned conditions had heavy background. Prolonged blocking
(48 hours) and adjustment of primary (1:5000) and secondary antibody dilution
(1:20000) did reduce the background, but no conclusive evidence could be obtained
from the blot (See Figure 5).
Figure 5: Testing EDG-3 (42kd) antibody with negative and positive controls.
NS: Negative Control Sample PS: Positive Control Sample
Two sets of controls were loaded on the membrane. No leukemia cell line samples were
loaded.
A: Blocking overnight. Primary antibody (anti-EDG-3) dilution 1:1000, incubated for 1
hour, secondary antibody dilution 1:10000.
B: Blocking for 48 hours. Primary antibody (anti-EDG-3) dilution 1:5000, incubated for
1 hour, secondary antibody dilution 1:20000.
C: Beta-actin. Blocking overnight. Primary antibody (anti-Beta-actin) dilution 1:1000,
incubated for 1 hour, secondary antibody dilution 1:10000.
Exposure time: 5 seconds.
20
EDG-8 antibody exhibited reduced binding affinity, with no bands being detected at
1:1000 primary antibody dilution (Other conditions remained the same as
aforementioned). Increased incubation time (overnight at 4
0
C) did not result in
observable bands, and increase of primary antibody concentration (1:500, 1:200 and
1:100) seemed to have led to unspecific binding, especially at the concentration of
1:100, but no bands of expected size were detected (See Figure 6).
Figure 6: Testing EDG-8 (46kd) antibody with negative and positive controls
NS: Negative Control Sample PS: Positive Control Sample
A: Primary antibody (anti-EDG-8) dilution 1:1000, incubating overnight at 4
0
C
B: Primary antibody (anti-EDG-8) dilution 1:500, incubating overnight.
C: Primary antibody (anti-EDG-8) dilution 1:200, incubating overnight.
D: Primary antibody (anti-EDG-8) dilution 1:100, incubating overnight.
Exposure time: 10 seconds.
21
Discussion
In summary, the current project has shown that three S1P receptors, EDG-1, EDG-5 and
EDG-6 are expressed in four T-ALL cell lines, while being unable to provide sufficient
evidence for the expression of EDG-3 and EDG-8. Attempts have been made to
manipulate certain key factors in experimental conditions, such as prolonging blocking
time from overnight to 48 hours, varying primary antibody concentration and incubation
time. Unfortunately these efforts have not enabled us to improve the quality of blots
remarkably. There exists the possibility of improving the blots by a more refined
adjustment of parameters, e.g., varying secondary antibody concentration / incubation
time, changing the total amount of sample loading, or using different type of membrane
(e.g., nitrocellulose membrane). But purchasing new antibodies from other
manufacturers would be more likely to put us in a better position to detect the expression
of EDG-3 and EDG-8.
EDG-1 has been shown to be important in cell proliferation, migration and cytoskeletal
organization in various cell lines (See introduction). However, these discoveries were
achieved in cell lines that are not leukemia-related, and thus may not be able to provide
illuminating information regarding the role of EDG-1 in leukemia. A more recent report
22
demonstrated that EDG-1 promotes lymphocyte egress from lymphoid organs by
overriding retention signals (Pham et al., 2008), which might have provided a link
between EDG-1 and leukemia, i.e., the expression of the S1P receptor EDG-1 could be
facilitating the egress of immature lymphocytes through constantly binding to S1P, a
molecule abundant in plasma.
EDG-5 has been widely studied but seems to exhibit a more controversial array of
effects in different cell lines. Conflicting reports on its role in regulating cell
proliferation, migration and angiogenesis have been published (Kon, et al., 1999; An et
al., 2000; Inoke et al., 2006). No research has been specifically designed to study the
function of EDG-5 in leukemia cell lines, and our current detection of EDG-5 expression
in T-ALL cell lines may provide initiatives for such studies in the future.
Since PCR data have long been acquired about the restricted expression of EDG-6 in
hematopoietic systems (Graler et al. 1998;Ishii et al. 2001), we anticipated an expression
of this receptor in T-ALL cell lines, and our western blot data confirmed our hypothesis.
Based on previous studies on the stimulatory role of EDG-6 on T cell proliferation and
cytokine secretion (Wang, et al., 2005), we further hypothesize that EDG-6 functions
similarly in T-ALL and promotes cell growth.
23
In light of previous data on the function of EDG-3 in HL-60 cells (Sato, et al., 1998) and
the mRNA expression of EDG-8 in human leukocytes, we also anticipated to detect both
EDG-3 and EDG-8 in T-ALL cells. However, our antibodies appeared ineffective during
the time of our experiments, and as of now, we are unable to provide convincing data for
their expression or lack of expression.
Based on the current study, future research could be directed to further profiling EDG
receptor expression in T-ALL, using a bigger panel of cell lines and more effective
antibodies. More importantly, we should make endeavor to establish a more in-depth
understanding of the signaling pathways mediated by different EDG receptors, as well as
the interactions and functions of these possible pathways. Combined, these studies are
expected to not only expand our general knowledge of leukemia, but also provide more
candidate targets for the development of new treatment of leukemia. One of the
foreseeable candidates, for example, would be an EDG-6 antagonist, which
competitively binds to EDG-6, while not inducing the regular signaling pathways
mediated by EDG-6. Some preliminary data from our lab have also shown that
sphingosine kinase 1 may be involved in EDG-6 dependent leukemia cell proliferation.
Therefore, sphingosine kinase inhibitors (e.g., dimethylsphingosine or safingol) could be
used individually or in combination with EDG-6 down-regulating agents (antisense or
siRNA) as a new potential candidate for drug testing.
24
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Abstract (if available)

Abstract The implication of S1P in cancer has attracted researchers from a wide spectrum of fields, but the relationship between S1P and leukemia has not been well studied, with limited data indicating a possible anti-apoptotic and pro-survival role of S1P that may lead to drug resistance. The current thesis aims at a characterization of the expression profile of S1P receptors in T-cell ALL leukemia using western blot analysis. It is hypothesized that one or more S1P receptors are expressed in a panel of T-ALL cell lines so that S1P-induced signals are transduced by these receptors. We expect that western blot analysis can detect the expression of these receptors. This project will lay the groundwork for the future research on S1P signaling pathways in T-ALL cell lines and may potentially provide more target molecules for drug development against T-cell ALL.

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Asset Metadata

Creator Zhang, Yue (author)

Core Title Characterizing the protein expression of S1P receptors in T-cell acute lymphoblastic leukemia (T-ALL) cell lines

School School of Dentistry

Degree Master of Science

Degree Program Craniofacial Biology

Publication Date 12/10/2008

Defense Date 10/31/2008

Publisher University of Southern California (original), University of Southern California. Libraries (digital)

Tag acute lymphoblastic leukemia,OAI-PMH Harvest,sphingosine 1 phosphate receptor,western blots

Language English

Contributor Electronically uploaded by the author (provenance)

Advisor Paine, Michael L. (committee chair), Maurer, Barry (committee member), Mosteller, Raymond (committee member), Zeichner-David, Margarita (committee member)

Creator Email alan_chen_1977@hotmail.com,yuez@usc.edu

Permanent Link (DOI) https://doi.org/10.25549/usctheses-m1905

Unique identifier UC178545

Identifier etd-Zhang-2530 (filename),usctheses-m40 (legacy collection record id),usctheses-c127-141625 (legacy record id),usctheses-m1905 (legacy record id)

Legacy Identifier etd-Zhang-2530.pdf

Dmrecord 141625

Document Type Thesis

Rights Zhang, Yue

Type texts

Source University of Southern California (contributing entity), University of Southern California Dissertations and Theses (collection)

Repository Name Libraries, University of Southern California

Repository Location Los Angeles, California

Repository Email cisadmin@lib.usc.edu