Introduction: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered to be an effective apoptosis inducer due to its selectivity for tumor cells. However, many cancer cells, especially metastatic cancer cells, often exhibit resistance to TRAIL because their apoptotic pathway is impaired or their pro-survival pathway is overactivated. TRAIL resistance is the main obstacle to current TRAIL therapy. Nowadays, ceramide analogs represent a new class of potential anticancer agents. Therefore, we hypothesized that disrupting pro-survival signaling with ceramide analogs would increase TRAIL-mediated apoptosis. Methods: MTT assay and flow cytometry were conducted to evaluate the synergistic effect of ceramide analog 5cc on TRAIL in metastatic colon cancer cells. Western blot was used to detect signaling proteins affected by 5cc. RNA interference was performed to analyze the effects of specific gene on 5cc-enhanced apoptosis. Results: Ceramide analog 5cc markedly enhanced TRAIL-induced apoptosis evidenced by increased propidium iodide/annexin V double-positive cells and PARP cleavage in SW620 and LS411N cells. At the molecular level, 5cc significantly reduced the expression of anti-apoptotic protein X-linked inhibitor of apoptosis protein (XIAP) through the activation of the c-Jun n-terminal kinase (JNK) pathway which is critically involved in sensitizing tumor cells to TRAIL/5cc combination. JNK-silenced cells exhibited a significant reversal of TRAIL/5cc-mediated apoptosis. Conclusion: Our data demonstrated that ceramide analog 5cc overcomes TRAIL resistance by enhancing JNK activation and repressing XIAP expression in metastatic colon cancer cells.

1.
Selvarajoo
K
.
A systems biology approach to overcome trail resistance in cancer treatment
.
Prog Biophys Mol Bio
.
2017
;
128
:
142
54
.
2.
Wang
S
,
El-Deiry
WS
.
Trail and apoptosis induction by tnf-family death receptors
.
Oncogene
.
2003
;
22
(
53
):
8628
33
.
3.
MacFarlane
M
.
Trail-induced signalling and apoptosis
.
Toxicol Lett
.
2003
139
2–3
89
97
.
4.
Forero-Torres
A
,
Infante
JR
,
Waterhouse
D
,
Wong
L
,
Vickers
S
,
Arrowsmith
E
.
Phase 2, multicenter, open-label study of tigatuzumab (cs-1008), a humanized monoclonal antibody targeting death receptor 5, in combination with gemcitabine in chemotherapy-naive patients with unresectable or metastatic pancreatic cancer
.
Cancer Med
.
2013
;
2
(
6
):
925
32
.
5.
Herbst
RS
,
Eckhardt
SG
,
Kurzrock
R
,
Ebbinghaus
S
,
O'Dwyer
PJ
,
Gordon
MS
.
Phase i dose-escalation study of recombinant human apo2l/trail, a dual proapoptotic receptor agonist, in patients with advanced cancer
.
J Clin Oncol
.
2010
;
28
(
17
):
2839
46
.
6.
Stuckey
DW
,
Shah
K
.
Trail on trial: preclinical advances in cancer therapy
.
Trends MOL MED
.
2013
;
19
(
11
):
685
94
.
7.
Maksimovic-Ivanic
D
,
Stosic-Grujicic
S
,
Nicoletti
F
,
Mijatovic
S
.
Resistance to trail and how to surmount it
.
Immunol Res
.
2012
52
1–2
157
68
.
8.
Rahman
M
,
Pumphrey
JG
,
Lipkowitz
S
.
The trail to targeted therapy of breast cancer
.
Adv Cancer Res
.
2009
;
103
:
43
73
.
9.
Cummins
JM
,
Kohli
M
,
Rago
C
,
Kinzler
KW
,
Vogelstein
B
,
Bunz
F
.
X-linked inhibitor of apoptosis protein (xiap) is a nonredundant modulator of tumor necrosis factor-related apoptosis-inducing ligand (trail)-mediated apoptosis in human cancer cells
.
Cancer Res
.
2004
;
64
(
9
):
3006
8
.
10.
Bratton
SB
,
Cohen
GM
.
Death receptors leave a caspase footprint that smacs of xiap
.
Cell Death Differ
.
2003
;
10
(
1
):
4
6
.
11.
Bode
AM
,
Dong
Z
.
The functional contrariety of jnk
.
Mol Carcinog
.
2007
;
46
(
8
):
591
8
.
12.
Gkouveris
I
,
Nikitakis
NG
.
Role of jnk signaling in oral cancer: a mini review
.
Tumour Biol
.
2017
;
39
(
6
):
1010428317711659
.
13.
Oh
HL
,
Seok
JY
,
Kwon
CH
,
Kang
SK
,
Kim
YK
.
Role of mapk in ceramide-induced cell death in primary cultured astrocytes from mouse embryonic brain
.
Neurotoxicology
.
2006
;
27
(
1
):
31
8
.
14.
Brindley
GS
.
An implant for occluding arteries
.
J Physiol
.
1975
246
2
28P
30P
.
15.
Zhang
TH
,
Liu
JF
,
Zhang
Y
,
Li
YL
,
Lu
HT
,
Murata
NM
.
Ceramide induces apoptosis in human lung adenocarcinoma a549 cells through mitogen-activated protein kinases
.
Acta Pharmacol Sin
.
2007
;
28
(
3
):
439
45
.
16.
Qiu
L
,
Zhou
C
,
Sun
Y
,
Di
W
,
Scheffler
E
,
Healey
S
.
Paclitaxel and ceramide synergistically induce cell death with transient activation of egfr and erk pathway in pancreatic cancer cells
.
Oncol Rep
.
2006
;
16
(
4
):
907
13
.
17.
Kong
JY
,
Klassen
SS
,
Rabkin
SW
.
Ceramide activates a mitochondrial p38 mitogen-activated protein kinase: a potential mechanism for loss of mitochondrial transmembrane potential and apoptosis
.
Mol Cell Biochem
.
2005
278
1–2
39
51
.
18.
Stoica
BA
,
Movsesyan
VA
,
Knoblach
SM
,
Faden
AI
.
Ceramide induces neuronal apoptosis through mitogen-activated protein kinases and causes release of multiple mitochondrial proteins
.
Mol Cell Neurosci
.
2005
;
29
(
3
):
355
71
.
19.
Mahalingam
D
,
Keane
M
,
Pirianov
G
,
Mehmet
H
,
Samali
A
,
Szegezdi
E
.
Differential activation of jnk1 isoforms by trail receptors modulate apoptosis of colon cancer cell lines
.
Br J Cancer
.
2009
;
100
(
9
):
1415
24
.
20.
Maundrell
K
,
Antonsson
B
,
Magnenat
E
,
Camps
M
,
Muda
M
,
Chabert
C
.
Bcl-2 undergoes phosphorylation by c-jun n-terminal kinase/stress-activated protein kinases in the presence of the constitutively active gtp-binding protein rac1
.
J Biol Chem
.
1997
;
272
(
40
):
25238
42
.
21.
Chen
L
,
Meng
Y
,
Guo
X
,
Sheng
X
,
Tai
G
,
Zhang
F
.
Gefitinib enhances human colon cancer cells to trail-induced apoptosis of via autophagy- and jnk-mediated death receptors upregulation
.
Apoptosis
.
2016
;
21
(
11
):
1291
301
.
22.
Ogretmen
B
.
Sphingolipid metabolism in cancer signalling and therapy
.
Nat Rev Cancer
.
2018
;
18
(
1
):
33
50
.
23.
Liu
Q
,
Li
X
,
Bao
YS
,
Lu
J
,
Li
H
,
Huang
Z
.
Chemical synthesis and functional characterization of a new class of ceramide analogues as anti-cancer agents
.
Bioorg Med Chem
.
2019
;
27
(
8
):
1489
96
.
24.
Yang
D
,
Wang
S
,
Brooks
C
,
Dong
Z
,
Schoenlein
PV
,
Kumar
V
.
Ifn regulatory factor 8 sensitizes soft tissue sarcoma cells to death receptor-initiated apoptosis via repression of flice-like protein expression
.
Cancer Res
.
2009
;
69
(
3
):
1080
8
.
25.
Li
X
,
Zeng
X
,
Sun
J
,
Li
H
,
Wu
P
,
Fung
KP
.
Imperatorin induces mcl-1 degradation to cooperatively trigger bax translocation and bak activation to suppress drug-resistant human hepatoma
.
Cancer Lett
.
2014
348
1–2
146
55
.
26.
Muhlenbeck
F
,
Haas
E
,
Schwenzer
R
,
Schubert
G
,
Grell
M
,
Smith
C
.
Trail/apo2l activates c-jun nh2-terminal kinase (jnk) via caspase-dependent and caspase-independent pathways
.
J Biol Chem
.
1998
;
273
(
49
):
33091
8
.
27.
Herr
I
,
Wilhelm
D
,
Meyer
E
,
Jeremias
I
,
Angel
P
,
Debatin
KM
.
Jnk/sapk activity contributes to trail-induced apoptosis
.
Cell Death Differ
.
1999
;
6
(
2
):
130
5
.
28.
Corazza
N
,
Jakob
S
,
Schaer
C
,
Frese
S
,
Keogh
A
,
Stroka
D
.
Trail receptor-mediated jnk activation and bim phosphorylation critically regulate fas-mediated liver damage and lethality
.
J Clin Invest
.
2006
;
116
(
9
):
2493
9
.
29.
Kudriaeva
AA
,
Belogurov
AA
.
Proteasome: a nanomachinery of creative destruction
.
Biochemistry
.
2019
84
Suppl 1
S159
S192
.
30.
Guo
N
,
Peng
Z
.
Mg132, a proteasome inhibitor, induces apoptosis in tumor cells
.
Asia-Pac J Clin Onco
.
2013
;
9
(
1
):
6
11
.
31.
Chauhan
D
,
Li
G
,
Hideshima
T
,
Podar
K
,
Mitsiades
C
,
Mitsiades
N
.
Jnk-dependent release of mitochondrial protein, smac, during apoptosis in multiple myeloma (mm) cells
.
J Biol Chem
.
2003
;
278
(
20
):
17593
6
.
32.
Tang
G
,
Minemoto
Y
,
Dibling
B
,
Purcell
NH
,
Li
Z
,
Karin
M
.
Inhibition of jnk activation through nf-kappab target genes
.
Nature
.
2001
;
414
(
6861
):
313
7
.
33.
Ashkenazi
A
,
Pai
RC
,
Fong
S
,
Leung
S
,
Lawrence
DA
,
Marsters
SA
.
Safety and antitumor activity of recombinant soluble apo2 ligand
.
J Clin Invest
.
1999
;
104
(
2
):
155
62
.
34.
Wagner
KW
,
Punnoose
EA
,
Januario
T
,
Lawrence
DA
,
Pitti
RM
,
Lancaster
K
.
Death-receptor o-glycosylation controls tumor-cell sensitivity to the proapoptotic ligand apo2l/trail
.
Nat Med
.
2007
;
13
(
9
):
1070
7
.
35.
Stolfi
C
,
Pallone
F
,
Monteleone
G
.
Molecular targets of trail-sensitizing agents in colorectal cancer
.
Int J Mol Sci
.
2012
;
13
(
7
):
7886
901
.
36.
Trivedi
R
,
Mishra
DP
.
Trailing trail resistance: novel targets for trail sensitization in cancer cells
.
Front Oncol
.
2015
;
5
:
69
.
37.
Senbabaoglu
F
,
Cingoz
A
,
Kaya
E
,
Kazancioglu
S
,
Lack
NA
,
Acilan
C
.
Identification of mitoxantrone as a trail-sensitizing agent for glioblastoma multiforme
.
Cancer Biol Ther
.
2016
;
17
(
5
):
546
57
.
38.
de Wilt
LH
,
Kroon
J
,
Jansen
G
,
de Jong
S
,
Peters
GJ
,
Kruyt
FA
.
Bortezomib and trail: a perfect match for apoptotic elimination of tumour cells
.
Crit Rev Oncol Hemat
.
2013
;
85
(
3
):
363
72
.
39.
Feng
C
,
Han
X
,
Chi
L
,
Sun
J
,
Gong
F
,
Shen
Y
.
Synthesis, characterization, and in vitro evaluation of trail-modified, cabazitaxel -loaded polymeric micelles for achieving synergistic anticancer therapy
.
J Biomat Sci-Polym E
.
2018
;
29
(
14
):
1729
44
.
40.
Gross
S
,
Rahal
R
,
Stransky
N
,
Lengauer
C
,
Hoeflich
KP
.
Targeting cancer with kinase inhibitors
.
J Clin Invest
.
2015
;
125
(
5
):
1780
9
.
41.
Kim
I
,
Byeon
HJ
,
Kim
TH
,
Lee
ES
,
Oh
KT
,
Shin
BS
.
Doxorubicin-loaded porous plga microparticles with surface attached trail for the inhalation treatment of metastatic lung cancer
.
Biomaterials
.
2013
;
34
(
27
):
6444
53
.
42.
Fugio
LB
,
Coeli-Lacchini
FB
,
Leopoldino
AM
.
Sphingolipids and mitochondrial dynamic
.
Cells
.
2020
;
9
(
3
):
581
.
43.
Saddoughi
SA
,
Garrett-Mayer
E
,
Chaudhary
U
,
O'Brien
PE
,
Afrin
LB
,
Day
TA
.
Results of a phase II trial of gemcitabine plus doxorubicin in patients with recurrent head and neck cancers: serum C-ceramide as a novel biomarker for monitoring response
.
Clin Cancer Res
.
2011
;
17
(
18
):
6097
105
.
44.
Deng
X
,
Yin
X
,
Allan
R
,
Lu
DD
,
Maurer
CW
,
Haimovitz-Friedman
A
.
Ceramide biogenesis is required for radiation-induced apoptosis in the germ line of c. Elegans
.
Science
.
2008
;
322
(
5898
):
110
5
.
45.
Li
RZ
,
Wang
XR
,
Wang
J
,
Xie
C
,
Wang
XX
,
Pan
HD
.
The key role of sphingolipid metabolism in cancer: new therapeutic targets, diagnostic and prognostic values, and anti-tumor immunotherapy resistance
.
Front Oncol
.
2022
;
12
:
941643
.
46.
Thapa
B
,
Kc
R
,
Uludağ
H
.
Trail therapy and prospective developments for cancer treatment
.
J Control Release
.
2020
;
326
:
335
49
.
47.
Paschall
AV
,
Zimmerman
MA
,
Torres
CM
,
Yang
D
,
Chen
MR
,
Li
X
.
Ceramide targets xiap and ciap1 to sensitize metastatic colon and breast cancer cells to apoptosis induction to suppress tumor progression
.
Bmc Cancer
.
2014
;
14
:
24
.
48.
Zhang
X
,
Li
B
,
Zhang
Y
,
Liu
J
.
Ceramide induces release of mitochondrial proapoptotic proteins in caspase-dependent and -independent manner in ht-29 cells
.
Sci China C Life Sci
.
2008
;
51
(
1
):
66
71
.
49.
Saraei
R
,
Soleimani
M
,
Movassaghpour Akbari
AA
,
Farshdousti Hagh
M
,
Hassanzadeh
A
,
Solali
S
.
The role of xiap in resistance to tnf-related apoptosis-inducing ligand (trail) in leukemia
.
Biomed Pharmacother
.
2018
;
107
:
1010
9
.
50.
Allensworth
JL
,
Aird
KM
,
Aldrich
AJ
,
Batinic-Haberle
I
,
Devi
GR
.
Xiap inhibition and generation of reactive oxygen species enhances trail sensitivity in inflammatory breast cancer cells
.
Mol Cancer Ther
.
2012
;
11
(
7
):
1518
27
.
51.
Hu
WH
,
Johnson
H
,
Shu
HB
.
Tumor necrosis factor-related apoptosis-inducing ligand receptors signal nf-kappab and jnk activation and apoptosis through distinct pathways
.
J Biol Chem
.
1999
;
274
(
43
):
30603
10
.
52.
Yabu
T
,
Shiba
H
,
Shibasaki
Y
,
Nakanishi
T
,
Imamura
S
,
Touhata
K
.
Stress-induced ceramide generation and apoptosis via the phosphorylation and activation of nsmase1 by jnk signaling
.
Cell Death Differ
.
2015
;
22
(
2
):
258
73
.
53.
Verheij
M
,
Bose
R
,
Lin
XH
,
Yao
B
,
Jarvis
WD
,
Grant
S
.
Requirement for ceramide-initiated sapk/jnk signalling in stress-induced apoptosis
.
Nature
.
1996
;
380
(
6569
):
75
9
.
54.
Bai
L
,
Smith
DC
,
Wang
S
.
Small-molecule smac mimetics as new cancer therapeutics
.
Pharmacol Ther
.
2014
;
144
(
1
):
82
95
.
55.
Tsai
YT
,
Huang
HC
,
Kao
ST
,
Chang
TT
,
Cheng
CY
.
Neuroprotective effects of alpinia oxyphylla miq against mitochondria-related apoptosis by the interactions between upregulated p38 mapk signaling and downregulated jnk signaling in the subacute phase of cerebral ischemia-reperfusion in rats
.
Am J Chin Med
.
2022
;
50
(
8
):
2057
83
.
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