Detailed molecular fingerprinting of four new anaplastic thyroid carcinoma cell lines and their use for verification of RhoB as a molecular therapeutic target.
BACKGROUND
Anaplastic thyroid carcinoma (ATC) is a highly aggressive carcinoma in need of therapeutic options. One critical component of drug discovery is the availability of well-characterized cell lines for identification of molecular mechanisms related to tumor biology and drug responsiveness. Up to 42% of human thyroid cancer cell lines are redundant or not of correct tissue origin, and a comprehensive analysis is currently nonexistent. Mechanistically, RhoB has been identified as a novel molecular target for ATC therapy.


OBJECTIVE
The aim was to develop four ATC cell lines detailing genetic, molecular, and phenotypic characteristics and to test five classes of drugs on the cell lines to determine whether they inhibited cell proliferation in a RhoB-dependent fashion.


METHODS
Four cell lines were derived from ATC tumors. Short tandem DNA repeat and mutational status of the originating tumors and cell lines were performed along with molecular and phenotypic characterizations. Compounds were tested for growth inhibition and ability to up-regulate RhoB.


RESULTS
Cell line authenticity was confirmed by DNA short tandem repeat analysis. Each proved unique regarding expression of thyroid markers, oncogene status, amplified and deleted genes, and proliferative growth rates. FTI-277, GGTI-286, lovastatin, romidepsin, and UCN-01 up-regulated RhoB and inhibited cell proliferation in a dose-responsive fashion with only romidepsin and FTI-277 being RhoB dependent.


CONCLUSIONS
Molecular descriptions of thyroid lines were matched to the originating tumors, setting a new standard for cell line characterization. Furthermore, suppressed RhoB is implicated as a molecular target for therapy against ATC because five classes of drugs up-regulate RhoB and inhibit growth dose-responsively.
Marlow
Laura A
LA
Mayo Clinic, Department of Cancer Biology, 4500 San Pablo Road, Jacksonville, Florida 32224, USA.
D'Innocenzi
Jaclyn
J
Zhang
Yilin
Y
Rohl
Stephen D
SD
Cooper
Simon J
SJ
Sebo
Thomas
T
Grant
Clive
C
McIver
Bryan
B
Kasperbauer
Jan L
JL
Wadsworth
J Trad
JT
Casler
John D
JD
Kennedy
Pamela W
PW
Highsmith
W Edward
WE
Clark
Orlo
O
Milosevic
Dragana
D
Netzel
Brian
B
Cradic
Kendall
K
Arora
Shilpi
S
Beaudry
Christian
C
Grebe
Stefan K
SK
Silverberg
Marc L
ML
Azorsa
David O
DO
Smallridge
Robert C
RC
Copland
John A
JA
D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D016172:DNA Fingerprinting; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005434:Flow Cytometry; D005819:Genetic Markers; D006801:Humans; D018895:Microsatellite Repeats / Q000235:genetics*; D009154:Mutation; D065646:Thyroid Carcinoma, Anaplastic; D013964:Thyroid Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D014162:Transfection; D020743:rhoB GTP-Binding Protein / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*
D000970:Antineoplastic Agents; D004273:DNA, Neoplasm; D005819:Genetic Markers; D020743:rhoB GTP-Binding Protein; D004121:Dimethyl Sulfoxide
J Clin Endocrinol Metab. 2003 Jul;88(7):3021-6
12843138
J Clin Endocrinol Metab. 2009 Dec;94(12):5001-9
19850689
Endocr Relat Cancer. 2005 Sep;12(3):615-29
16172195
Cancer Cell Int. 2003 Jul 11;3(1):10
12890289
Thyroid. 2001 Aug;11(8):725-31
11525264
Endocr Relat Cancer. 2009 Mar;16(1):17-44
18987168
Cancer Res. 2009 Feb 15;69(4):1536-44
19208833
Cancer Res. 2005 Nov 15;65(22):10199-207
16288007
Nat Rev Cancer. 2010 Jun;10(6):441-8
20448633
Cancer Chemother Pharmacol. 2006 Oct;58(4):460-70
16435154
J Clin Endocrinol Metab. 2008 Nov;93(11):4331-41
18713817
Methods. 2001 Dec;25(4):402-8
11846609
Nat Rev Drug Discov. 2009 Jul;8(7):547-66
19568282
Endocrinology. 2003 Sep;144(9):3852-9
12933658
Nat Rev Cancer. 2001 Nov;1(2):162-8
11905808
Cancer Res. 2004 May 15;64(10):3629-37
15150122
Cancer Cell. 2007 Feb;11(2):175-89
17292828