Research On The Effect And Mechanism Of Isoliquirtigenin On Ovarian Cancer Metastasis

Objective:To investigate the metastasis of the inhibitory effects of isoliquirtigenin on ovarian cancer.Methods:In vitro test:a.MTT assay was used to detect the effects of isoliquirtigenin on the growth of ovarian cancer cell lines OVCAR5,SKOV3,ES2 and TOV21G.b.Three concentrations of 1,5,10 ?M in different concentrations of isoliquirtigenin were used.Transwell chamber experiment and wound healing scratch test were used to test whether isoliquirtigenin can inhibit the motility of ovarian cancer cells.c.Using a matrix-coated Transwell chamber to detect whether isoliquirtigenin can inhibit the invasive ability of ovarian cancer cells.d.The effect of isoliquirtigenin on the morphology of EMT in ovarian cancer cells was observed by inverted microscope.e.The effect of isoliquirtigenin on the expression of EMT-related marker protein was detected by Western blotting.f.The effect of isoliquirtigenin on Racl activity was examined using the GLISA kit.g.Detection of the effects of isoliquirtigenin Erkl/2 and Src protein expression by Western blotting.h.EMT PCR Array and Q-PCR were used to screen and verify the effects of isoliquirtigenin on EMT-related genes and transcription factors.(2)In vivo:a.Construction of human ovarian cancer luciferase cell line SKOV3/Luc.b.Establish a model of ovarian cancer metastasis in nude mice by intraperitoneal injection.c.Bioluminescence was used to observe the growth and metastasis of ovarian cancer in vitro in nude mice and to measure the body weight of each nude mouse.d.Continued administration until the natural death of nude mice,to study the effect of isoliquirtigenin on the survival of nude mice.e.Anatomy,observe the metastasis of the tumor in the abdominal cavity,and collect the tumor for immunohistochemical analysis.f.Anatomy,the tumor was detected by Q-PCR for changes in mRNA levels of related signaling pathway proteins and EMT transcription factors.Results:1.High concentration of isoliquirtigenin inhibited the growth of ovarian cancer cells,and low concentration of isoliquirtigenin had on effect on ovarian cancer cells.2.After ovarian cancer cells SKOV3 and OVCAR5 were treated with 1OjiM isoliquirtigenin for 72h,the morphology of the cells changed obviously,and the cell morphology changed from slender interstitial morphology to pebble-like epithelial morphology.3.IOjiM isoliquirtigenin can significantly inhibit the migration and invasion of ovarian cancer cells SKOV3 and OVCAR5.4.After treatment with ovarian cancer cells SKOV3 and OVCAR5 for 72 h,the expression of Vimentin and N-cadherin proteins in EMT could be down-regulated,and the expression of E-cadherin was up-regulated,and EMT transcription factor ZEB1 was down-regulated.5.10?M isoliquirtigenin can reduce Racl activity in ovarian cancer cells.6.10?M isoliquirtigenin can down-regulate the expression of p-Erk1/2 and p-Src protein,and has no effect on Erkl/2 and Src protein expression.7.In vivo animal experiments,the model of ovarian cancer peritoneal metastasis was successful.Compared with the control group,isoliquirtigenin can prolong the survival of ovarian cancer animals.The results of immunohistochemistry and Q-PCR were consistent with the in vitro results.Conclusion:Isoliquirtigenin reverses the EMT process of ovarian cancer through Racl-MEK1/Src signaling pathway,and ZEB1 transcription factor regulates this process,thereby inhibiting the metastasis of ovarian cancer.