Arsenic trioxide-induced apoptosis of Hep-2 cell line through modulating intracellular glutathione (GSH) level.
BACKGROUND
Since 1970s, a Chinese study group at Harbin Medical University First Hospital discovered the anticancer effect of arsenic trioxide (As2O3), it has become evident that apoptotic effects of As2O3 are not restricted to APL cells but also can be observed in other malignant cells in vitro, including non-APL acute myeloid leukemia cells, myeloma cells, and chronic myeloid leukemia cells, as well as various solid-tumor cells, such as esophageal, prostate, and ovarian carcinomas and neuroblastoma cells.


OBJECTIVE
To investigated if As2O3 could induce cell death and apoptosis of Hep-2 cells, a laryngeal squamous cell carcinoma (LSCC) cell line.


METHODS
Trypan blue exclusion assay, LDH release assay and cytometric method were used for the measurements of cell death and apoptosis. We measured intracellular GSH, ROS and mitochondrial membrane potential to explore the mechanisms of cell death and apoptosis induced by As2O3 in Hep-2 cells.


RESULTS
Trypan-blue-positive cells and the release of LDH into medium induced by As2O3 increased in a dose- and time-dependent manner. The rates of apoptosis increased induced by As2O3 in a dose- and time-dependent manner. In order to elucidate the mechanisms of cell death and apoptosis induced by As2O3 in Hep-2 cells through GSH, we found that As2O3 induced the decrease of intracellular GSH, increase of ROS and loss of mitochondrial membrane potential. And pretreatment of BSO, an inhibitor of GSH biosynthesis, depleted partly intracellular GSH and increased trypan-blue-positive cells, the release of LDH, apoptosis, ROS and loss of mitochondrial membrane potential. However, pretreatment of GSH had resistance to the changes to some extent as described above.


CONCLUSIONS
As2O3-induced apoptosis of Hep-2 cell line through modulating intracellular GSH level.
Cheng
Baohua
B
Jining Medical College, Jining City, Shandong Province, China. apple9red@yahoo.com.cn
Yang
Xinxin
X
An
Liangxiang
L
Gao
Bo
B
Liu
Xia
X
D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000494:pharmacology* / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects*; D000077237:Arsenic Trioxide; D001152:Arsenicals / Q000008:administration & dosage / Q000494:pharmacology* / Q000627:therapeutic use*; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy* / Q000473:pathology*; D045744:Cell Line, Tumor; D005978:Glutathione / Q000378:metabolism*; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects* / Q000378:metabolism*; D007822:Laryngeal Neoplasms / Q000188:drug therapy* / Q000473:pathology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D010087:Oxides / Q000008:administration & dosage / Q000494:pharmacology* / Q000627:therapeutic use*; D017382:Reactive Oxygen Species
D000970:Antineoplastic Agents; D001152:Arsenicals; D010087:Oxides; D017382:Reactive Oxygen Species; D005978:Glutathione; D000077237:Arsenic Trioxide