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ObjectiveTo study the possible molecular mechanism of baicalein （BAI）-mediated focal adhesion kinase （FAK） in the regulation of phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway to inhibit the proliferation and migration of gastric cancer HGC-27 cells. MethodThe gastric epithelial GES-1 cells and gastric cancer HGC-27 cells were respectively treated with BAI （0， 5， 15， 25， and 50 μmol·L-1） for 48 h， and then methyl thiazolyl tetrazolium （MTT） assay was adopted to detect effect of BAI on cell proliferation. Western blot （WB） was employed to detect the expression of FAK and the proteins related to epithelial-mesenchymal transition （EMT） and PI3K signaling pathway after intervention with different concentrations of BAI. The HGC-27 cells stably overexpressing FAK were constructed with lentivirus-mediated transfection technique， and the transfection of FAK was detected through WB and green fluorescent protein （GFP）. The cells were divided into empty vector （NC） group， BAI group， FAK overexpression group， and BAI-treated FAK overexpression group， and cell proliferation activity was detected by MTT assay. The colony formation and cell migration were observed via colony formation assay and Transwell migration assay， respectively. The expression of proteins involved in EMT and PI3K signaling pathways were detected by Western blot. ResultCompared with the NC group， BAI （15， 25 and 50 μmol·L-1） inhibited the proliferation of HGC-27 cells in a dose-dependent manner （P<0.05， P<0.01） while did not affect that of GES-1 cells. BAI （5， 15 and 25 μmol·L-1） down-regulated the expression level of p-FAK （P<0.05， P<0.01）. Compared with NC group， FAK overexpression group showed up-regulated expression level of FAK in HGC-27 cells. The HGC-27 cells in both NC group and FAK overexpression group had green fluorescence. Compared with NC group， BAI inhibited the growth， colony formation， and migration， while FAK overexpression promoted those of HGC-27 cells. The treatment of FAK overexpression group with BAI inhibited the enhancement of cell proliferation and migration （P<0.05）. WB showed that compared with NC group， BAI （15， 25 μmol·L-1） significantly up-regulated the expression of E-cadherin protein and down-regulated that of Vimentin， Snail， p-PI3K， and p-Akt protein in HGC-27 cells （P<0.05， P<0.01）. Compared with NC group， FAK overexpression group showed down-regulated expression of E-cadherin， up-regulated expression of p-FAK， Vimentin， and Snail， and increased ratios of p-FAK/FAK， p-PI3K/PI3K and p-Akt/Akt （P<0.05）. This phenomenon would be reversed after BAI treatment. ConclusionBAI can affect the proliferation and migration of gastric cancer HGC-27 cells by mediating FAK to regulate PI3K/Akt signaling pathway.
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Objective To explore the effect of silencing focal adhesion kinase (FAK) gene on the proliferation and migration of human tongue squamous cell carcinoma cell lines CAL-27.Methods The siRNA interference technology was used to complete the construction of FAK siRNA by transient transfection.CAL-27 cells were divided into the experiment group,the negative control group and blank control group.The expressions of FAK mRNA and protein were detected by qPCR and Western blotting respectively.The cell proliferation ability was tested by using MTT assay.The cell migration ability was detected by using Transwell chamber assay.Results The expression level of FAK mRNA and protein in experiment group was lower than that in blank control group and negative control group,the difference was significant (PinRNA ＜ 0.01,Pprolein ＜ 0.05).Compared with negative control group and blank control group,the proliferation ability of cells in experiment group was obviously decreased at 48 h and 72 h (P ＜ 0.01).Transwell chamber migration assay showed,the average number of migrating cells in experimnet group group was lower than that in blank control group and negative control group,the difference was significant (P ＜ 0.05).Conclusions FAK gene silencing can significantly inhibit the proliferation ability and migration ability of human tongue squamous cell carcinoma cell lines CAL-27.
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Objective To investigate the relationships between expression of FAK and Prostate Carcinoma (PC)morbidity. Methods By surgery,get cancer tissue and para-carcinama tissue from 60 cases PC.To detect FAK expression by immuno-histochemical.To extract total RNA by Trizol.To detect the FAK mRNA by RT-PCR,and analysis these data.Results FAK expression level in cancer tissue was higher than that in para-carcinama tissue.There was statistical difference between them (χ2=72.55,P<0.01).mRNA expression levels of FAK in cancer tissue showed significant higher than the levels in para-carcinama tissue (t=30.51,P<0.01).According to lymphatic metastasis,the expression positive cases of FAK in pN0M0 classification were lower than these in pN3M1 classification,and mRNA expression levels of FAK were the same re-sults.There were clearly statistical distinctive (t=25.43,P<0.01).Conclusion The expression of FAK in PC cells was as-sociated with tumor invasion.
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Objective To investigate the effect of Biglycan and FAK signal pathway on the proliferation of colon cancer cells in vitro and its possible mechanisms.Methods Biglycan expression vector was constructed and transfected into the colon cancer cell line HCT116.FAK inhibitor was used for cell treatment as well.Cells were divided into 5 groups:control group(HCT116),control group transfected with empty plasmid(Vector),con-trol group with empty plasmid transfected and inhibitor treatment(Vector+PF-562271),group transfected with Biglycan expression vector(Biglycan),group with Biglycan expression vector transfected and inhibitor treatment ( Biglycan+PF-562271) .Treatment duration was 24 hours.The expressions of FAK,p-FAK,PCNA and p53 were detected by Western Blot.The proliferation of cells was detected by MTT.Results The overexpression of Biglycan significantly promoted the proliferation of HCT116 and the phosphorylation of FAK(P<0.01).It signif-icantly up-regulated PCNA and down-regulated p53(P<0.01).The FAK inhibitor PF-562271 treatment could obviously inhibit the proliferation of HCT116,and the regulation of Biglycan on the expression of p-FAK, PCNA.p53 proteins was reversed(P<0.01).Conclusion Biglycan regulates the proliferation of colon cancer cells by promoting the activation of FAK signal pathway.
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Studies have proved that gastrin plays key roles in the development and progression of gastroenteric tumor. After gastrin combined with the tumor cell surface receptors, some genetic transcription and expression were adjusted through certain signal transduction pathways, resulting in the development and growth of gastrointestinal carcinoma. Effective treatment method would been founded about gastroenteric tumor through the studies of mechanism of action and signal transduction pathways of gastrin in gastroenteric tumor.
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Background and purpose:PTEN mutation has been found in 20%-40% of malignant gliomas.The common mutant epidermal growth factor receptor(EGFR vⅢ)was reported to coexpress in PTEN-deficient EGFR-expressing tumor.PTEN has been shown to interact directly with FAK and reduce its tyrosine phosphorylation levels to inhibit cell invasion.The invasion of glioma cells with EGFRvⅢ expression and PTEN deficiency is increased.This study was to observe whether PTEN inhibits glioma cell invasion even in the presence of strong pro-invasive signals provided by constitutive EGFR activity.Methods:U87?EGFR cells were transfected with pcDNA3.1 constructs encoding PTEN and the cells invasion levels were detected by transwell invasion assay.The expression of FAK was detected by immunoblotting.FAK expression vector was transfected into U87?EGFR-wtPTEN cells and the change of cells invasion was documented.Results:PTEN and PTEN(G129E)could inhibit cell invasion induced by EGFRvⅢ.PTEN and PTEN(G129E)could decrease the FAK phosphorylation at Tyr397.Over expression of FAK in U87?EGFR-PTEN abrogated PTEN-induced down-regulation of the phosphorylation status of FAK and rescued cell invasion.Conclusions:PTEN could inhibit cell invasion induced by EGFRvⅢ by dephosphorylating FAK.