In vitro synergistic cytotoxicity of gemcitabine and pemetrexed and pharmacogenetic evaluation of response to gemcitabine in bladder cancer patients.
The present study was performed to investigate the capability of gemcitabine and pemetrexed to synergistically interact with respect to cytotoxicity and apoptosis in T24 and J82 bladder cancer cells, and to establish a correlation between drug activity and gene expression of selected genes in tumour samples. The interaction between gemcitabine and pemetrexed was synergistic; indeed, pemetrexed favoured gemcitabine cytotoxicity by increasing cellular population in S-phase, reducing Akt phosphorylation as well as by inducing the expression of a major gemcitabine uptake system, the human equilibrative nucleoside transporter-1 (hENT1), and the key activating enzyme deoxycytidine kinase (dCK) in both cell lines. Bladder tumour specimens showed an heterogeneous gene expression pattern and patients with higher levels of dCK and hENT1 had better response. Moreover, human nucleoside concentrative transporter-1 was detectable only in 3/12 patients, two of whom presented a complete response to gemcitabine. These data provide evidence that the chemotherapeutic activity of the combination of gemcitabine and pemetrexed is synergistic against bladder cancer cells in vitro and that the assessment of the expression of genes involved in gemcitabine uptake and activation might be a possible determinant of bladder cancer response and may represent a new tool for treatment optimization.
Mey
V
V
Division of Pharmacology and Chemotherapy, Department of Internal Medicine, University of Pisa, 55, Via Roma, I-56100, 56126, Pisa, Italy.
Giovannetti
E
E
De Braud
F
F
Nannizzi
S
S
Curigliano
G
G
Verweij
F
F
De Cobelli
O
O
Pece
S
S
Del Tacca
M
M
Danesi
R
R
D000283:Administration, Intravesical; D000328:Adult; D000368:Aged; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003841:Deoxycytidine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000494:pharmacology; D003842:Deoxycytidine Kinase / Q000037:antagonists & inhibitors / Q000235:genetics; D004176:Dipyridamole / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D033721:Equilibrative Nucleoside Transporter 1 / Q000187:drug effects / Q000235:genetics; D018691:Excitatory Amino Acid Antagonists; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D005971:Glutamates / Q000008:administration & dosage* / Q000494:pharmacology; D006147:Guanine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000494:pharmacology; D006801:Humans; D008297:Male; D008875:Middle Aged; D009367:Neoplasm Staging; D000068437:Pemetrexed; D051026:Phosphoribosylglycinamide Formyltransferase / Q000037:antagonists & inhibitors / Q000235:genetics; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000378:metabolism; D016196:S Phase / Q000187:drug effects; D013868:Thioinosine / Q000031:analogs & derivatives / Q000494:pharmacology; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors / Q000235:genetics; D016896:Treatment Outcome; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy* / Q000378:metabolism*; D000093542:Gemcitabine
D033721:Equilibrative Nucleoside Transporter 1; D018691:Excitatory Amino Acid Antagonists; D005971:Glutamates; D000068437:Pemetrexed; D003841:Deoxycytidine; D013868:Thioinosine; D006147:Guanine; D004176:Dipyridamole; D013940:Thymidylate Synthase; D051026:Phosphoribosylglycinamide Formyltransferase; D003842:Deoxycytidine Kinase; D051057:Proto-Oncogene Proteins c-akt; C001789:4-nitrobenzylthioinosine; D000093542:Gemcitabine