The Experimental Study On The Effect Of The Gekko Swinhonis Anti-neoplasm Active Component On The HepG2Cells In Vitro

Objective This study was to investigate the effects of the Gekko Swinhonis anti-neoplasm active component（GSAAC） on the proliferation, migration and apoptosisof HepG2cells. The aim of our study was to provide datum for the further researchand development of the Gekko Swinhonis anti-neoplasm active component and toevaluate the action and possible mechanism on anti-tumour in vitro.Materials and methods Insitute of Biochemistry and Cell Biology, CAS provided usHepG2cells. The HepG2were routinely grown in RPMI-1640medium containing10%fetal bovine serum. The HepG2were devided into different concentration of thefresh GEKKO SWINHONIS anti-neoplasm active component groups、cisplatingroup and control group. HepG2cells were treated with GSAAC(25mg·L^-1、50mg·L-1、100mg·L^-1、200mg·L^-1、400mg·L^-1). The HepG2proliferation was detected by theMTT assay; the expressions of PCNA of HepG2was detected byimmunohistochemistry; The HepG2migration and invasion was observed byTranswell assay after treatment; Hoechst33342fluorescence staining and TUNELstaining were used to observe the apoptosis morphological changes of HepG2cells;Early apoptosis and cell cycle distribution were assessed by flow cytometry. Alldatum was analyze by the SPSS13.0software.Results1. The effects of the GSAAC on the proliferation of HepG2cells: TheGSAAC(25、50、100、200、400mg·L^-1) groups can significantly inhibit the proliferation of HepG2cells. The inhibiting rate of GSAAC(100、200、400mg·L^-1)groups higher than50%level（24h-72h） in a dose-and time-dependentmanner（P<0.01or P<0.05）. The OD value of MTT shows, The inhibiting rate ofGSAAC(200、400mg·L^-1) groups higher than DDP group （48h-72h）（P<0.01orP<0.05）.2. After treated with the GSAAC(25、50、100、200、400mg·L^-1) groups for48hours,the proliferation of GSAAC(25、50、100、200、400mg·L^-1) groups were（67.31±5.45%;64.36±4.56%;60.87±5.03%;53.85±3.48%;49.38±3.55%） lower thanthe control group （72.28±6.09%） in a dose-dependent manner（P<0.01or P<0.05）.3. The effects of the GSAAC on the migration and invasion of HepG2cells: Aftertreated with the GSAAC(25、50、100、200、400mg·L^-1) groups for24hours. theGSAAC (25、50、100、200、400mg·L^-1) groups can inhibit the migration and invasionof HepG2cells. The number of migration and invasion of HepG2cells of theGSAAC(100、200、400mg·L^-1) groups were lower than the control group. Thenumber of migration and invasion of HepG2cells of the GSAAC(200、400mg·L^-1)groups significantly lower than the DDP group（P<0.01）.4. The effects of the GSAAC on the apoptosis of HepG2cells:（1）The GSAAC caninduce the apoptosis morphological changes of HepG2cells by Hoechst33342fluorescence staining and TUNEL staining.（2）The GSAAC can induce the earlyapoptosis of HepG2cells. The early apoptosis rate of HepG2cells of the GSAAC(25、100、400mg·L^-1) groups were （3.94±0.43%、6.25±1.12%、7.98±2.08%）, the DDPgroup was （6.71±1.34%） higher than the control group （2.36±0.23%）.（P<0.01）. in adose-dependent manner.5. The effects of the GSAAC on the cell-cycle distribution of HepG2cells:Comparedwith the control group, The GSAAC(25、100、400mg·L^-1) groups and the DDP groupcan block HepG2cells to go to G2phase from S phase（P<0.01）.Conclusion The GSAAC can inhibit the proliferation、migration and invision ofHepG2cells. Parts of the intrinsic mechanism might relate to the blockage of HepG2cells to go to G2phase from S phase thus to inhibit the proliferation、migration and the induction the early apoptosis of HepG2cells.