Mechanism of Liushenwan and Realgar in Inducing Apoptosis and DNA Damage in Human Endometrial Cancer JEC Cells / 中国实验方剂学杂志

ABSTRACT

Objective:Compare the anti-tumor effect and mechanism of Liushenwan and realgar (As4S4) on human endometrial cancer cells JEC. Method:The release of As in Liushenwan and As4S4 was measured by atomic absorption spectrometry. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used for cell proliferation, and cell migration was measured by Transwell assay. Flow cytometry and Western Blot were used to determine apoptosis and DNA damage. Result:The dissolution of As in Liushenwan was 17.4%, and that of As4S4 was only 1.6% according to atomic absorption assay. With the same content of As, compared with the As4S4 group, the cell viability in the 3，10 mg·L-1 Liushenwan groups was decreased (P<0.05), the early apoptosis rate was significantly increased in 0.25，0.5，1 mg·L-1 Liushenwan groups (P<0.05), the rate of cell migration was decreased in 1 mg·L-1 Liushenwan group (P<0.05), the expressions of cleaved cysteinyl aspartate-specific proteinase-3 (cleaved Caspase-3), cleaved cysteinyl aspartate-specific proteinase-7 (cleaved Caspase-7), cleaved poly ADP-ribose polymerase (cleaved PARP), phosphorylated histone (p-H2AX), phosphorylation of checkpoint kinase 2 (p-CHK2) and ataxia-telangiectasia mutated (ATM) were increased in 1 mg·L-1 Liushenwan group (P<0.05), while the expression of phosphorylation of ataxia-telangiectasia mutated rad3-related (ATR) was decreased in 1 mg·L-1 Liushenwan group (P<0.05), with no significant changes in the expressions of cysteinyl aspartate-specific proteinase-3 (Caspase-3) and cysteinyl aspartate-specific proteinase-7 (Caspase-7). Conclusion:With the same content of As, both Liushenwan and As4S4 could inhibit JEC cell proliferation and migration, and induce cell apoptosis and DNA damage. Liushenwan has a stronger effect than As4S4. It is suggested that there are other components in Liushenwan with an anti-tumor effect in cooperation with As.