Identification of thymidylate synthase as a potential therapeutic target for lung cancer.
BACKGROUND
Thymidylate synthase (TS), a key enzyme in the de novo synthesis of thymidine, is an important chemotherapeutic target for malignant tumours including lung cancer. Although inhibition of TS has an antiproliferative effect in cancer cells, the precise mechanism of this effect has remained unclear.


METHODS
We examined the effects of TS inhibition with an RNA interference-based approach. The effect of TS depletion on the growth of lung cancer cells was examined using colorimetric assay and flow cytometry.


RESULTS
Measurement of the enzymatic activity of TS in 30 human lung cancer cell lines revealed that such activity differs among tumour histotypes. Almost complete elimination of TS activity by RNA interference resulted in inhibition of cell proliferation in all tested cell lines, suggestive of a pivotal role for TS in cell proliferation independent of the original level of enzyme activity. The antiproliferative effect of TS depletion was accompanied by arrest of cells in S phase of the cell cycle and the induction of caspase-dependent apoptosis as well as by changes in the expression levels of cyclin E and c-Myc. Moreover, TS depletion induced downregulation of the antiapoptotic protein X-linked inhibitor of apoptosis (XIAP), and it seemed to activate the mitochondrial pathway of apoptosis.


CONCLUSIONS
Our data provide insight into the biological relevance of TS as well as a basis for clinical development of TS-targeted therapy for lung cancer.
Takezawa
K
K
Department of Medical Oncology, Kinki University School of Medicine, 377-2 Ohno-higashi, Osaka-Sayama, Osaka 589-8511, Japan.
Okamoto
I
I
Tsukioka
S
S
Uchida
J
J
Kiniwa
M
M
Fukuoka
M
M
Nakagawa
K
K
D000230:Adenocarcinoma / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D017209:Apoptosis; D018287:Carcinoma, Large Cell / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D018288:Carcinoma, Small Cell / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D002294:Carcinoma, Squamous Cell / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D002453:Cell Cycle / Q000235:genetics; D002455:Cell Division / Q000235:genetics; D045744:Cell Line, Tumor; D019927:Cyclin E / Q000235:genetics; D003600:Cytosol / Q000378:metabolism; D017353:Gene Deletion; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D015151:Immunoblotting; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D008928:Mitochondria / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D034622:RNA Interference; D016196:S Phase / Q000235:genetics; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism
D019927:Cyclin E; D016271:Proto-Oncogene Proteins c-myc; D013940:Thymidylate Synthase; D053148:Caspase 3