ABSTRACT
Objective:To investigate the regulatory effect of lumbrukinase (LBK) on the gastric cancer SGC7901cells, and to clarify its mechanism.Methods:The SGC7901cells in the logarithmic growth phase were selected and divided into control group and 2, 4, 8U·mL-1 LBK groups.MTT assay was used to detect the inhibitory rates of proliferation of SGC7901cells in various groups in vitro at different time (24, 48and 72h) .Cell scratch assay was used to detect the migration abilities of the SGC7901cells in vitro in various groups.Flow cytometry was used to determine the apoptotic rates of SGC7901cells and the pencentages of cells at different cell cycles in various groups.The expression levels of Bcl-2, Bax, and caspase-3in the SGC7901cells in various groups were detected by Western blotting method.Results:The results of MTT assay showed that compared with control group, the inhibitory rates of SGC7901cells in different doses of LBK groups after treated for 24, 48and 72hwere increased (P＜0.01) .The cell scratch assay results showed that compared with control group, the migration distances of SGC7901cells in4and 8U·mL-1 LBK groups were increased significantly (P＜0.01) .The flow cytometry results showed that compared with control group, the apoptotic rates of SGC7901cells in 4and 8U·mL-1 LBK groups were increased significantly (P＜0.01) ;the percentages of cells in G1and S phases were decreased (P＜0.01) ;the percentages of cells in G2phase were increased (P＜0.01) .The results of Western blotting method showed that compared with control group, the Bcl-2protein expression level in the SGC7901cells in 8U·mL-1 LBK group was decreased (P＜0.05) ;the Bax and caspase-3protein expression levels were increased (P＜0.05) .Conclusion:LBK can inhibit the proliferation and migration abilities of SGC7901cells in vitro and induce the apoptosis;its mechanism is achieved through the regulation of expression levels of Bcl-2, Bax, and caspase-3proteins.
ABSTRACT
Objective: To investigate the regulatory effect of lumbrukinase CI.RK) on the gastric cancer SGC7901 Cells, and to clarify its mechanism Methods: The SGC7901 cells in the logarithmic growth phase were selected and divided into control group and 2, 4 , 8 U • ml. I.RK groups. MTT assay was used to detect the inhibitory rates of proliferation of SGC7901 cells in various groups in vitro at different time (24, 48 and 72 h). Cell scratch assay was used to detect the migration abilities of the SGC7901 cells in vitro in various groups. Flow cytometry was used to determine the apoptotic rates of SGC7901 cells and the pencentages of cells at different cell cycles in various groups. The expression levels of Bcl-2, Rax, and caspase-3 in the SGC790I cells in various groups were detected by Western blotting method. Results: The results of MTT assay showed that compared with control group, the inhibitory rates of SGC7901 cells in different doses of I.RK groups after treated for 24, 48 and 72 h were increased ( P<0.01). The cell scratch assay results showed that compared with control group, the migration distances of SGC790I cells in 4 and 8 U • mL"1 I.RK groups were increased significantly ( P
ABSTRACT
Aim To establish a new,multi-parameter,real time and qualitative cell migration evaluation method.Methods Lung cancer cell line A549 was cultured on the glass bottom dish.After treated with different dosages of berberine or Rg3 for 24 hours,several scratching lines at the same dimension were made and observed by Living Cell Working Station.8 observation areas were selected randomly and imaged continuously for 24 hours.Transferred Area(TA),Transferred Distance(TD),Single Cell Transferred Speed(SCTS)and the Cell Division Number among defined area(CDN)were analyzed after getting sequence images.Results The focus stage and the incubation system were sufficient to keep cell proliferation and made it possible for long term observation.Berberine at 25 μmol·L~(-1) and 12.5 μmol·L~(-1) could inhibit the migration of A549(P<0.01).The analysis results of TA,TD,SCTS and CDN were basically coincident.Rg3 at 0.1 mmol·L~(-1) could inhibit SCTS and promote CDN in 6,12 and 24 h(P<0.01),while decrease both TA and TD in 24 hs.Conclusion The method is sensitive,rapid and simple to be applied in the research of tumor metastasis,wound healing and inflammatory response with real time,in-situ and multi-parameters.