Induction of apoptosis by organotin compounds in vitro: neuronal protection with antisense oligonucleotides directed against stannin.
Immortalized cell lines and primary neuronal cultures were used to characterize the selective toxicity of trimethyltin (TMT),triethyltin (TET) and tributyltin (TBT). TBT and TET were cytotoxic at similar concentrations in the immortalized cell lines tested; the 50% toxic concentration (TC50) was 1 to 11 microM. In contrast, immortalized cell lines varied considerably in their sensitivity to TMT, with sensitive cell lines (neuroblastomas, T-, B-cell lines) showing TC50 values of 2 to 8 microM, whereas insensitive cells (NIH-3T3 fibroblast, HTB-14 glioma, TC-7 kidney cells) had TC 50 values > 100 microM. Primary neuronal cell cultures were very sensitive to organotins (TC50 values, 1-10nM), and showed patterns of selective toxicity with respect to neuronal and glial cells. Because organotin toxicity evolves over 24 to 48 hr. we determined whether these compounds induced apoptosis in primary cultures. TMT increased (P < .05) the fraction of apoptotic cells 6 and 12 hr after treatment with TMT at TC50 concentrations. Prior studies suggested that a protein, stannin, was localized in cells sensitive to organotins. Stannin was expressed in several TMT-sensitive cell lines (PC12, T, B cells) and in primary neurons in culture. Stannin was absent in the resistant HTB-14 glioma cell line. The role of stannin in mediating TMT toxicity in primary cultures was investigated by blocking stannin expression with specific antisense oligonucleotides. Treatment of primary cultures with antisense oligonucleotides for 48 hr before and during TMT treatment significantly protected neurons from the neurotoxic and apoptotic effects of TMT. This effect was not observed with scrambled oligonucleotide controls. Thus, TMT may induce apoptosis in sensitive cells, which is partly mediated by stannin. Based on the available data we conclude that stannin expression is necessary, but not sufficient for TMT toxicity.
Thompson
T A
TA
Department of Pharmacology, Pennsylvania State University College of Medicine, Hershey, PA, USA.
Lewis
J M
JM
Dejneka
N S
NS
Severs
W B
WB
Polavarapu
R
R
Billingsley
M L
ML
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D006624:Hippocampus / Q000187:drug effects; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D009479:Neuropeptides / Q000494:pharmacology*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D009947:Organotin Compounds / Q000494:pharmacology*; D051381:Rats; D014267:Triethyltin Compounds / Q000494:pharmacology; D014298:Trimethyltin Compounds / Q000494:pharmacology
D009479:Neuropeptides; D016376:Oligonucleotides, Antisense; D009947:Organotin Compounds; D014267:Triethyltin Compounds; D014298:Trimethyltin Compounds; C046488:trimethyltin; C075692:stannin; C031575:triethyltin