Adenoviral transduction of TESTIN gene into breast and uterine cancer cell lines promotes apoptosis and tumor reduction in vivo.

PURPOSE The human TESTIN (TES) gene is a putative tumor suppressor gene in the fragile chromosomal region FRA7G at 7q31.1/2 that was reported to be altered in leukemia and lymphoma cell lines. In this report, we investigated the effect of TES gene expression in vivo to evaluate a possible role of TES gene in human cancer. EXPERIMENTAL DESIGN We have analyzed the expression of TES gene in a panel of 25 breast tumors and 17 cell lines of breast, colon, and uterine cancers. Furthermore, to evaluate the potential of TES gene therapy, we studied the effects of adenoviral TES transduction (Ad-TES) in cell lines with undetectable TES expression (T47D and MES-SA) as well as in MCF-7 cell line where TES expression is normal. RESULTS Twenty-five percent of primary breast tumor samples as well as the breast cancer cell line T47D and the uterine sarcoma cell line MES-SA were negative or displayed low levels of TES. After TES restoration by Ad-TES transduction, T47D and MES-SA cell lines underwent apoptosis. Furthermore, TES expression significantly reduced the tumorigenic potential of both T47D and MES-SA in nude mice, whereas the untreated cells and Ad-GFP-infected cells showed tumor growth in vivo. The TES-positive cell line control (MCF-7) was not affected by TES expression and did not show a reduction of tumorigenicity in nude mice after infection with Ad-TES. CONCLUSIONS Ad-TES expression inhibit the growth of breast and uterine cancer cells lacking of TES expression through caspase-dependent and caspase-independent apoptosis, respectively, suggesting that Ad-TES infection should be explored as a therapeutic strategy.

[1]  M. Lisanti,et al.  Caveolin-1, a putative tumour suppressor gene. , 2001, Biochemical Society transactions.

[2]  P. Vandenabeele,et al.  Mitochondrial intermembrane proteins in cell death. , 2003, Biochemical and biophysical research communications.

[3]  Alan G. Porter,et al.  Caspase-3 Is Required for DNA Fragmentation and Morphological Changes Associated with Apoptosis* , 1998, The Journal of Biological Chemistry.

[4]  Haojie Huang,et al.  Fish mapping of YAC clones at human chromosomal band 7q31.2: Identification of YACS spanning FRA7G within the common region of LOH in breast and prostate cancer , 1998, Genes, chromosomes & cancer.

[5]  P. Briand,et al.  Relationship between tumorigenicity, in vitro invasiveness, and plasminogen activator production of human breast cell lines. , 1990, European journal of cancer.

[6]  H. Iwase,et al.  [Breast cancer]. , 2006, Nihon rinsho. Japanese journal of clinical medicine.

[7]  J. Engelman,et al.  Genes encoding human caveolin‐1 and ‐2 are co‐localized to the D7S522 locus (7q31.1), a known fragile site (FRA7G) that is frequently deleted in human cancers , 1998, FEBS letters.

[8]  X. Sastre,et al.  Uterine sarcomas: the Curie Institut experience. Prognosis factors and adjuvant treatments. , 1999, Gynecologic oncology.

[9]  T. Le Adjuvant pelvic radiotherapy for uterine carcinosarcoma in a high risk population. , 2001, European journal of surgical oncology : the journal of the European Society of Surgical Oncology and the British Association of Surgical Oncology.

[10]  T. Macalma,et al.  Molecular Characterization of Human Zyxin* , 1996, The Journal of Biological Chemistry.

[11]  S. Scherer,et al.  Detailed deletion mapping with a refined physical map of 7q31 localizes a putative tumor suppressor gene for breast cancer in the region of MET. , 1996, Oncogene.

[12]  E. Tobias,et al.  The TES gene at 7q31.1 is methylated in tumours and encodes a novel growth-suppressing LIM domain protein , 2001, Oncogene.

[13]  Chaohua Yan,et al.  Apoptosis in the absence of caspase 3 , 2001, Oncogene.

[14]  K. Mimori,et al.  Characterization of the human TESTIN gene localized in the FRA7G region at 7q31.2. , 2000, Genomics.

[15]  G. Kroemer,et al.  Apoptosis-inducing factor (AIF): caspase-independent after all , 2004, Cell Death and Differentiation.

[16]  R. Scopes,et al.  Analysis of Proteins , 1998 .

[17]  R.K. Scopes,et al.  Analysis of Proteins , 1998 .

[18]  R. Aqeilan,et al.  Interleukin 2‐Bax: a novel prototype of human chimeric proteins for targeted therapy , 1999, FEBS letters.

[19]  D W SMITHERS,et al.  Clinical Cancer Research , 1941, Lancet.

[20]  D. Grossman,et al.  Rapid induction of mitochondrial events and caspase-independent apoptosis in Survivin-targeted melanoma cells , 2004, Oncogene.

[21]  Hyun-Ok Yang,et al.  Mechanism of taxol‐induced apoptosis in human SKOV3 ovarian carcinoma cells , 2004, Journal of cellular biochemistry.

[22]  A. Coutts,et al.  TES is a novel focal adhesion protein with a role in cell spreading , 2003, Journal of Cell Science.

[23]  T. Köcher,et al.  The conformational state of Tes regulates its zyxin-dependent recruitment to focal adhesions , 2003, The Journal of cell biology.

[24]  K. Kinzler,et al.  The Genetic Basis of Human Cancer , 1997 .

[25]  S. Grösch,et al.  G protein-independent G1 cell cycle block and apoptosis with morphine in adenocarcinoma cells: involvement of p53 phosphorylation. , 2003, Cancer research.

[26]  S. Agrawal,et al.  Survivin inhibition induces human neural tumor cell death through caspase‐independent and ‐dependent pathways , 2001, Journal of neurochemistry.