Improvement of sensitivity to platinum compound with siRNA knockdown of upregulated genes in platinum complex-resistant ovarian cancer cells in vitro.
It is known that some cancers show platinum complex resistance and that others show platinum complex sensitivity among ovarian cancers. Oxaliplatin (cis-[oxalato[trans-l-1, 2-diamino-cyclohexane] platinum[II]]; l-OHP), an active anti-cancer agent consisting of platinum, inhibits RNA synthesis and results in cytostatic effects. We investigated the difference between an oxaliplatin-resistant ovarian cancer cell line, KFR, and an oxaliplatin-sensitive ovarian cancer cell line, KF-1, using DNA microarray analysis. The oxaliplatin-resistant cell line, KFR, was established by using KF-1 cells derived from human serous cystadenocarcinoma of the ovary. Acquisition of platinum resistance in human ovarian cancer cells thus appeared to be related mainly to the expression of gamma-glutamylcysteine synthetase (gamma-GCS), topo II and metallothionein (hMT) genes, and partly to that of topo I and glutathione S-transferase--pi (GST-pi) genes, in addition to a decrease in platinum accumulation. KFR cells had 8.5- and 24.7-fold higher mRNA levels of gamma-glutamylcysteine synthetase (gamma-GCS), and topo II genes than KF-1 cells, while KFR had only a slight increase in the glutathione S-transferase--pi (GST-pi) mRNA level as compared with KF-1. In comparison of the gene expressions between KFR and KF-1 ovarian cancer cell lines, tubulin-specific chaperone E (TBCE) and CBP/p300-interacting transactivator (CITED2) were overexpressed in KFR compared to KF-1. These genes are overexpressed in MKN74, an oxaliplatin-resistant gastric cancer cell line, compared to MKN28, an oxaliplatin-sensitive gastric cancer cell line. TBCE is 13-fold increased in KFR cells compared to KF-1 cells. CBP/p300-interacting transactivator is increased 2-fold in KFR cells compared to KF-1 cells. The siRNA directed to the TBCE gene and CBP/p300-interacting transactivator gene enhanced the cytotoxicity of diplatin to the platinum-resistant ovarian cancer cell line KFR. These results show that the TBCE gene and CBP/p300 gene have potential as multidrug-resistant genes. It is necessary to check the effect of siRNA to influx or exflux. It has potential to enhance the effect of anti-cancer agents to resistant cancer cells, so we will proceed to develop an inhibitor of these TBCE and CBP/p300 proteins.
Department of Nuclear Engineering and Management, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan. yanagie@n.t.u-tokyo.ac.jp
Hironobu
H
Yanagie
Tomoyuki
T
Hisa
Aya
A
Ogata
Ayaka
A
Miyazaki
Yasumasa
Y
Nonaka
Teturo
T
Nishihira
Isao
I
Osada
Takayuki
T
Sairennji
Hirotaka
H
Sugiyama
Yoshitaka
Y
Furuya
Yosinori
Y
Kidani
Shinichi
S
Takamoto
Hiroyuki
H
Takahashi
Masazumi
M
Eriguchi
D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D018284:Cystadenocarcinoma, Serous / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D004250:DNA Topoisomerases, Type II / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D005721:Glutamate-Cysteine Ligase / Q000235:genetics; D051549:Glutathione S-Transferase pi / Q000235:genetics; D006801:Humans; D018832:Molecular Chaperones / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D009944:Organoplatinum Compounds / Q000494:pharmacology*; D010051:Ovarian Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D000077150:Oxaliplatin; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics; D013274:Stomach Neoplasms / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics; D014162:Transfection; D015854:Up-Regulation
D000970:Antineoplastic Agents; C486381:CITED2 protein, human; D018832:Molecular Chaperones; D009944:Organoplatinum Compounds; D034741:RNA, Small Interfering; D012097:Repressor Proteins; C469270:TBCE protein, human; D015534:Trans-Activators; D000077150:Oxaliplatin; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi; D004250:DNA Topoisomerases, Type II; D005721:Glutamate-Cysteine Ligase