Morphological and biochemical changes induced by arsenic trioxide in neuroblastoma cell lines.
Arsenic trioxide has recently been shown to inhibit growth and induce apoptosis in a variety of hematologic malignancies, but very little is known about its effects on solid tumors and especially on neuroblastoma cells that have self-differentiating characteristics. To demonstrate the growth inhibition induced in neuroblastoma cells (the SH-SY5Y and SK-N-AS cell line) and acute promyelocytic leukemia cells (HL-60) by arsenic trioxide (As2O3), the viable cell numbers were counted after trypan blue staining. Apoptosis was assessed by the cell morphology, by flow cytometry with annexin-V staining, and by Western blot analysis for the apoptosis-related proteins (bcl-2 and PARP). To decide the dose for the clinical application of As2O3, normal peripheral blood lymphocytes were also examined. The growth and survival of the SH-SY5Y and SK-N-AS cells were markedly inhibited by As2O3 treatment at a 3 microM concentration before the changes of the normal lymphocytes were observed. The apoptotic cells showed a shrunken cell nucleus, and an increase in the number and balloon-like swelling of the mitochondria at 72 h after the As2O3 was added. Apoptosis of the annexin-V-positive cell proportion in the neuroblastoma cell lines was increased with increasing the exposure time and the concentration of As2O3, just like the HL-60 cells. Bcl-2 downregulation and PARP degradation were also noted all the cell lines, but these changes were not statistically significant among the 3 cell lines. Taken together, these results indicate that As2O3 is an excellent candidate as a therapeutic agent for the treatment of neuroblastoma.
Ryu
Kyung-Ha
KH
Department of Pediatrics and Ewha Medical Research Institute, Ewha Woman's University College of Medicine, Seoul, Korea.
Woo
So-Youn
SY
Lee
Mi-Young
MY
Jung
Yun-Jae
YJ
Yoo
Eun-Sun
ES
Seoh
Ju-Young
JY
Kie
Jeong-Hae
JH
Shin
Hee-Young
HY
Ahn
Hyo-Seop
HS
D017304:Annexin A5 / Q000096:biosynthesis; D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects*; D000077237:Arsenic Trioxide; D001152:Arsenicals / Q000494:pharmacology* / Q000627:therapeutic use; D002470:Cell Survival / Q000187:drug effects; D024161:Collagen Type XI / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D018922:HL-60 Cells; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism / Q000648:ultrastructure; D009447:Neuroblastoma / Q000188:drug therapy / Q000378:metabolism* / Q000648:ultrastructure; D010087:Oxides / Q000494:pharmacology* / Q000627:therapeutic use; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis
D017304:Annexin A5; D000970:Antineoplastic Agents; D001152:Arsenicals; C499319:COL11A2 protein, human; D024161:Collagen Type XI; D010087:Oxides; D019253:Proto-Oncogene Proteins c-bcl-2; D000077237:Arsenic Trioxide