Advances in Bioscience and Biotechnology, 2014, 5, 73-80 ABB
http://dx.doi.org/10.4236/abb.2014.51011 Published Online January 2014 (http://www.scirp.org/journal/abb/ )
OPEN ACCESS
Cell-type specific and non-redundant anti-proliferative
effects of shRNA-mediated Galpha12- and Galpha13
knockdown in lung cancer cell lines
Thomas R. H. Büch
1,2*#
, Marius Grzelinski
1,2,3#
, Olaf Pinkenburg
1,2,4
, Thomas Gudermann
5,6
,
Achim Aigner
1,2*
1
Rudolf-Boehm-Institute for Pharmacology and Toxicology, University of Leipzig, Leipzig, Germany
2
Biochemical Pharmacological Institute/bpc, Philipps-University Marburg, Marburg, Germany
3
Present Address: Targos Molecular Pathology GmbH, Kassel, Germany
4
Present Address: Institute for Immunology, Philipps-University Marburg, Marburg, Germany
5
Walther Straub Institute of Pharmacology and Toxicology, University of Munich (LMU), Munich, Germany
6
Comprehensive Pneumology Center Munich (CPC-M), German Center for Lung Research, Munich, Germany
Email:
*
achim.aigner@medizin.uni-leipzig.de
Received 13 November 2013; revised 24 December 2013; accepted 13 January 2014
Copyright © 2014 Thomas R. H. Büch et al. This is an open access article distributed under the Creative Commons Attribution Li-
cense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. In
accordance of the Creative Commons Attribution License all Copyrights © 2014 are reserved for SCIRP and the owner of the intel-
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ABSTRACT
In small cell lung cancer cells, various autocrine sti-
muli lead to the parallel activation of Gq/11 and G
12/13
proteins. The contribution of the Gq/11-PLC-β cas-
cade to the mitogenic effects in SCLC cells is well es-
tablished, but the relevance of G
12/13
signaling is less
explored. While in prostate and breast cancer, G
12/13
activation has been shown previously to promote in-
vasiveness without being involved in cellular prolife-
ration, previous data from our group indicate anti-
proliferative effects of G
12/13
knockdown in small cell
lung cancer (SCLC) cells. To further investigate the
role of G
12/13
-dependent signaling in lung tumor cells,
we employed shRNA-mediated targeting of Gα
12
,
Gα
13
, or both, in SCLC and NSCLC cell lines. Lenti-
viral expression of shRNAs resulted in specific Gα
12
and Gα
13
knockdown. Of note, upon single knock-
down of one family member, no counter-upregulation
of the other one was observed. Interestingly, inhibi-
tion of proliferation was cell line dependent. In cell
lines where knock-down led to antiproliferation, sin-
gle knockdown of either Gα
12
or Gα
13
was sufficient
to impair proliferation and double knockdown of
Gα
12
and Gα
13
tended not to further increase an-
ti-proliferative effects. Likewise, when single knock-
down was insufficient for an inhibition of prolifera-
tion, no effects were observed in double knockdowns.
Taken together, these findings indicate that both Gα
12
and Gα
13
affect cellular proliferation individually and
interference with one family member is sufficient for
anti-tumor effects.
KEYWORDS
Galpha12; Galpha13; G
12/13
; Lung Cancer; SCLC;
NSCLC; Gene Knockdown
1. INTRODUCTION
Lung cancer represents the leading cause of cancer-re-
lated death in the industrialized countries. Cancers of the
lung are subdivided into different histological subtypes,
e.g., adenocarcinoma, squamous cell carcinoma, large
cell carcinoma and small cell carcinoma (SCLC). The
former three subtypes are often subsumed as non-small
cell carcinoma (NSCLC). Lung cancer of the SCLC type
shows a very aggressive behavior with early manifesta-
tion of metastases. Thus, local therapeutic options like
surgery are usually inapplicable in this tumor entity at
the time of diagnosis. While current chemotherapy regi-
mens in SCLC tumors clearly show clinical benefit, the
overall survival after 5 years is still only 7% - 12% [1].
In NSCLC, although the potential to develop metastases
is generally somewhat lower as compared to SCLC, dis-
semination is frequent at the time of diagnosis, necessi-
tating systemic chemotherapy. This clearly emphasizes
the need for novel treatment strategies based on the de-
*
Corresponding author.
#
These authors contributed equally to the work.