The investigation on the expression regulation and the role on chemosensitivity of a xenobiotic enzyme GSTP1 in lung cancer.
Grant-in-Aid for Scientific Research (C)
|Allocation Type||Single-year Grants |
Respiratory organ internal medicine
|Research Institution||Yokohama city university |
MATSUSE Takeshi Yokohama city university, medical center, professor, 医学部附属病院, 教授 (90199795)
NAKAJIMA Jun Tokyo university, school of medicine, associate professor, 医学部, 助教授 (90188954)
ITO Takaaki Kumamoto university, school of medicine, professor, 医学部, 教授 (70168392)
SHIRAI Akira Yokohama city university, medical center, associate professor, 医学部附属病院, 助教授 (40244488)
増田 道明 独協医科大学, 医学部, 教授 (80199702)
|Project Period (FY)
2002 – 2003
Completed (Fiscal Year 2003)
|Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2003: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
|Keywords||lung cencer / chemosensitivity / xenobiotic enzyme / GSTP1 / DNA methylation|
1.Induction of CpG island methylation of GSTP1 promoter in lung adenocarcinoma cells.
The CpG island of GSTP1 promoter was not methylated and GSTP1 was detected by immunoblotting in A549 lung adenocarcinoma cells. When we transduced the methylated oligonucleotides to these cells, methylation of the GSTP1 promoter and reduction of GSTP1 expression was induced.
2.Induction of CpG island methylation with DNA methyltransferases (DNMT)
It was observed that DNMT1, 3a, and 3b were all expressed in A549 cells. We cloned the cDNA of DNMT3b, and established a stable transfectant of DNMT3b from A549 cells. Now we check the methylation status of GSTP1 CpG island in these transfectants with bisulfite sequenting.
3.Association between GSTP1 expression and viability of lung-derived cells (including lung cancer cells)
We regulate the expression level of GSTP1 in human lung-derived fibroblast HFL-1 and A549 lung cancer cells, and investigated the role of GSTP1 on spontaneous apoptosis. Depletion of GSTP1 increased spontaneous apoptosis and necrosis in both kinds of cells. Although we investigated the mechanism of this phenomena, we found no relations between this phenomena and JNK pathway or intracellular concentration of glutathione.
4.Association between GSTP1 expression and chemosensitivity
As a pilot study, we carried out transfection of GSTP1 sense and antisense vectors to examine effects of GSTP1 on apoptosis induced by camptothecin in HeLa cells. Camptothecin-induced apoptosis was attenuated by transfection of GSTP1 sense vector in HeLa cells.
We also observed that GSTP1 depletion with siRNA increased camptothecin-induced cell death in A549 lung cancer cells.
Report (3 results)
Research Products (8 results)