Accepted Manuscript Title: AKT inhibition synergistically enhances growth-inhibitory effects of gefitinib and increases apoptosis in non-small cell lung cancer cell lines Author: M. Puglisi P. Thavasu A. Stewart J.S. de Bono M.E.R. O’Brien S. Popat J. Bhosle U. Banerji PII: DOI: Reference:
S0169-5002(14)00217-7 http://dx.doi.org/doi:10.1016/j.lungcan.2014.05.008 LUNG 4605
To appear in:
Lung Cancer
Received date: Revised date: Accepted date:
27-2-2014 7-5-2014 8-5-2014
Please cite this article as: Puglisi M, Thavasu P, Stewart A, de Bono JS, O’Brien MER, Popat S, Bhosle J, Banerji U, AKT inhibition synergistically enhances growth-inhibitory effects of gefitinib and increases apoptosis in non-small cell lung cancer cell lines, Lung Cancer (2014), http://dx.doi.org/10.1016/j.lungcan.2014.05.008 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
1
AKT inhibition synergistically enhances growth-
2
inhibitory effects of gefitinib and increases apoptosis
3
in non-small cell lung cancer cell lines
6
M Puglisi, 1 P Thavasu, 1 A Stewart 1JS de Bono, 2 MER O’Brien,
2
S Popat, 2 J Bhosle, 1 U Banerji
cr
1-2
5
7
1
8
Therapeutics & Clinical Studies, The Institute of Cancer
9
Research & The Royal Marsden NHS Foundation Trust, London, UK
11
2
12
Trust, London, UK
The Lung Unit, The Royal Marsden NHS Foundation
M
10
an
us
The Drug Development Unit, Divisions of Cancer
ip t
4
d
13
Address for correspondence:
15
Dr Udai Banerji
16
Division of Cancer Therapeutics & Division of Clinical Studies
17
The Institute of Cancer Research & The Royal Marsden
18
The Drug Development Unit, Sycamore House
19
Downs Road, Sutton, UK, SM2 5PT
20
tel: +44 20 8661 3984/fax: +44 (0) 20 8641 7979
21
[email protected]
Ac ce p
te
14
22 23
Word count: 1815
24 25
1 Page 1 of 33
26 27 28
ip t
29 30
cr
31
us
32 33
an
34 35
M
36 37
40
Objectives
te
Abstract
Ac ce p
39
d
38
41
EGFR inhibitors are ineffective against most EGFR wild-type
42
non-small cell lung cancer, for which novel treatment strategies
43
are needed. AKT signalling is essential for mediating EGFR
44
survival signals in NSCLC. We evaluated the combination of
45
gefitinib and two different AKT inhibitors, the allosteric
46
inhibitor AKTi-1/2 and the ATP-competitive pan-AKT
47
inhibitor AZD5363, in EGFR-mutant (HCC-827 and PC-9) and
2 Page 2 of 33
-wild-type (NCI-H522, NCI-H1651), non-small cell lung
49
cancer cell lines.
50
Materials and Methods
51
Drug interaction was studied in 2 EGFR mutant and 2 EGFR
52
wild-type non-small cell lung cancer cell lines by calculating
53
Combination Index (CI) using median effect analysis. The
54
effects on p-EGFR, p-ERK, p-AKT, p-S6 and apoptosis were
55
analysed by western blot.
56
Results
57
The combination of gefitinib and AKTi-1/2 or AZD5363
58
showed synergistic growth inhibition in all cell lines. CI values
59
for the combination of gefitinib and
60
(p=0.0048), 0.56 (p=0.036), 0.75 (p=0.13) and 0.64 (p=0.0003)
61
in NCI-H522, NCI-H1651, HCC-827 and PC-9 cell lines,
62
respectively; CI values of 0.45 (p=0.0087) and 0.22 (p
S0169-5002(14)00217-7 http://dx.doi.org/doi:10.1016/j.lungcan.2014.05.008 LUNG 4605
To appear in:
Lung Cancer
Received date: Revised date: Accepted date:
27-2-2014 7-5-2014 8-5-2014
Please cite this article as: Puglisi M, Thavasu P, Stewart A, de Bono JS, O’Brien MER, Popat S, Bhosle J, Banerji U, AKT inhibition synergistically enhances growth-inhibitory effects of gefitinib and increases apoptosis in non-small cell lung cancer cell lines, Lung Cancer (2014), http://dx.doi.org/10.1016/j.lungcan.2014.05.008 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
1
AKT inhibition synergistically enhances growth-
2
inhibitory effects of gefitinib and increases apoptosis
3
in non-small cell lung cancer cell lines
6
M Puglisi, 1 P Thavasu, 1 A Stewart 1JS de Bono, 2 MER O’Brien,
2
S Popat, 2 J Bhosle, 1 U Banerji
cr
1-2
5
7
1
8
Therapeutics & Clinical Studies, The Institute of Cancer
9
Research & The Royal Marsden NHS Foundation Trust, London, UK
11
2
12
Trust, London, UK
The Lung Unit, The Royal Marsden NHS Foundation
M
10
an
us
The Drug Development Unit, Divisions of Cancer
ip t
4
d
13
Address for correspondence:
15
Dr Udai Banerji
16
Division of Cancer Therapeutics & Division of Clinical Studies
17
The Institute of Cancer Research & The Royal Marsden
18
The Drug Development Unit, Sycamore House
19
Downs Road, Sutton, UK, SM2 5PT
20
tel: +44 20 8661 3984/fax: +44 (0) 20 8641 7979
21
[email protected]
Ac ce p
te
14
22 23
Word count: 1815
24 25
1 Page 1 of 33
26 27 28
ip t
29 30
cr
31
us
32 33
an
34 35
M
36 37
40
Objectives
te
Abstract
Ac ce p
39
d
38
41
EGFR inhibitors are ineffective against most EGFR wild-type
42
non-small cell lung cancer, for which novel treatment strategies
43
are needed. AKT signalling is essential for mediating EGFR
44
survival signals in NSCLC. We evaluated the combination of
45
gefitinib and two different AKT inhibitors, the allosteric
46
inhibitor AKTi-1/2 and the ATP-competitive pan-AKT
47
inhibitor AZD5363, in EGFR-mutant (HCC-827 and PC-9) and
2 Page 2 of 33
-wild-type (NCI-H522, NCI-H1651), non-small cell lung
49
cancer cell lines.
50
Materials and Methods
51
Drug interaction was studied in 2 EGFR mutant and 2 EGFR
52
wild-type non-small cell lung cancer cell lines by calculating
53
Combination Index (CI) using median effect analysis. The
54
effects on p-EGFR, p-ERK, p-AKT, p-S6 and apoptosis were
55
analysed by western blot.
56
Results
57
The combination of gefitinib and AKTi-1/2 or AZD5363
58
showed synergistic growth inhibition in all cell lines. CI values
59
for the combination of gefitinib and
60
(p=0.0048), 0.56 (p=0.036), 0.75 (p=0.13) and 0.64 (p=0.0003)
61
in NCI-H522, NCI-H1651, HCC-827 and PC-9 cell lines,
62
respectively; CI values of 0.45 (p=0.0087) and 0.22 (p
