RESUMO
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.
RESUMO
SATB1 plays a crucial role in the invasion and metastasis of breast cancer,and inhibition of SATB1 expression can effectively control breast cancer metastasis. In this study,homogeneous polysaccharides were isolated from Poria cocos and their sulfated derivatives were prepared to screen out the polysaccharide compositions with inhibitory effects on SATB1 expression. Smal-molecule components were removed from P. cocos by ethanol extraction,and P. cocos crude polysaccharide PPS was obtained by water extraction and ethanol precipitation. Then PPS was successively separated by DEAE Sepharose fast flow anion-exchange and Superdex-75 gel permeation chromatographic steps to give PPSW-1. The structure of PPSW-1 was identified and its sulfated derivatives were prepared. Then their inhibitory effects on human breast cancer MDA-MB-231 cells were investigated. A kind of polysaccharide,PPSW-1 with inhibitory effect on human breast cancer MDA-MB-231 cells,was obtained from P. cocos,with a relative molecular weight of 3. 06×104,and structure of 1,6-branched 1,3-α-D-galactan. PPSW-1 and its sulfated derivative Sul-W-1 showed good inhibitory effect on cells migration,and the water solubility of Sul-W-1 was better than that of PPSW-1. In addition,it was found that polysaccharide of P. cocos and its sulfated derivative can inhibit expression of SATB1. In this study,a kind of homogeneous polysaccharide with inhibitory effect on human breast cancer MDA-MB-231 cells was isolated from P. cocos,and its sulfated derivative with similar efficacy but better solubility was prepared,laying the foundation for the substance basis study of P. cocos.