The effect of small interfering RNA (siRNA) against the Bcl-2 gene on apoptosis and chemosensitivity in a canine mammary gland tumor cell line.
The aim of this study was to investigate whether downregulation of Bcl-2 expression by small interfering RNA (siRNA) against the canine Bcl-2 gene would enhance the apoptosis and sensitivity of a canine mammary gland tumor cell line (CF33) to doxorubicin. Transfections of CF33 with siRNA were performed using cationic liposomes. Sequence-specific downregulation of Bcl-2 expression was measured by semiquantitative RT-PCR and Western blot analysis. Total viable cells were determined by MTS assay and apoptotic cell rates were determined by the immunohistochemical analysis on ssDNA. Our data showed the siRNA downregulated Bcl-2 expression which increased apotosis and also increased the sensitivity of CF33 to doxorubicin. This study indicated that downregulation of Bcl-2 expression by siRNA would be useful as a new protocol to increase the effect of doxorubicin on treatment of canine mammary gland tumors, requiring a detailed evaluation of siRNA in vivo.
Nagamatsu
Kota
K
Department of Pathobiology, Nihon University School of Veterinary Medicine, Fujisawa, Kanagawa 252-8510, Japan.
Tsuchiya
Fumito
F
Oguma
Keisuke
K
Maruyama
Haruhiko
H
Kano
Rui
R
Hasegawa
Atsuhiko
A
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000235:genetics*; D045744:Cell Line, Tumor; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004317:Doxorubicin / Q000494:pharmacology*; D015972:Gene Expression Regulation, Neoplastic; D019254:Genes, bcl-2 / Q000235:genetics*; D015674:Mammary Neoplasms, Animal / Q000188:drug therapy* / Q000235:genetics* / Q000473:pathology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism
D000970:Antineoplastic Agents; D034741:RNA, Small Interfering; D004317:Doxorubicin