Effect of Hirsutine on Proliferation， Apoptosis， Metastasis， and Invasion of Human Cervical Cancer Ca Ski Cells / 中国实验方剂学杂志

ABSTRACT

ObjectiveTo investigate the effects of hirsutine on proliferation， apoptosis， metastasis， and invasion of human cervical cancer Ca Ski cells and its action mechanism. MethodThe cell proliferation was determined by cell counting kit-8 （CCK-8） assay and the cell apoptosis by Hoechst 33258 staining and flow cytometry. The mRNA expression levels of B cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and p53 were detected by real-time fluorescence quantitative polymerase chain reaction. The scratch test was conducted to detect the cell mobility， followed by the detection of cell invasion ability using a Transwell chamber. The contents of fibronectin （FN）， matrix metalloproteinase-2 （MMP-2）， and matrix metalloproteinase-9 （MMP-9） were measured by enzyme linked immunosorbent assay （ELISA）， and their protein expression levels were assayed by Western blot. ResultCompared with the control group， hirsutine inhibited the proliferation of Ca Ski cells in a dose- and time-dependent manner. The apoptosis rates and Bax and p53 mRNA expression levels in the 8.0， 16.0， 32.0 μmol·L-1 hirsutine groups rose （P<0.05， P<0.01）， while the Bcl-2 mRNA expression and phosphorylated Src （p-Src） and phosphorylated signal transducer and activator of transcription factor 3 （STAT3） protein expression declined （P<0.05， P<0.01）. Compared with the control group， the 1.0， 2.0， 4.0 μmol·L-1 hirsutine groups exhibited lowered cell mobility， number of transmembrane cells， FN， MMP-2 and MMP-9 contents， and hypoxia inducible factor-1α （HIF-1α）， Vimentin， and N-cadherin protein expression （P<0.05， P<0.01）， but elevated E-cadherin protein expression （P<0.01）. ConclusionHirsutine inhibits the proliferation， metastasis， and invasion of human cervical cancer Ca Ski cells and induces their apoptosis， which may be related to its regulation of Src/STAT3 and HIF-1α/epithelial mesenchymal transformation （EMT） signaling pathways.