Role And Mechanism Of TF In Anoikis Resistance Of Gastric Cancer

BackgroundsGastric cancer is the fourth most common malignant tumor in the world and the third most common cause of death from malignant tumors worldwide.Anoikis is a special form of programmed cell death induced by the loss of contact between cells and the extracellular matrix and other cells.Anoikis induces cell death through the traditional apoptotic pathway.However,recent studies have shown that tumor cells can resist this programmed death process.Once tumor cells begin to metastasize,they have the ability to resist apoptosis.Reports have shown that TF is highly expressed in various cancer species,and the high expression of TF is inversely correlated with the prognosis of many patients with malignant tumors.But the relationship between the expression of TF in gastric cancer and anoikis has not been studied.Objectives1.To study the expression of TF in normal gastric cell and gastric cancer cell lines with different degrees of differentiation;2.To investigate whether there is a difference in the expression of TF in gastric cancer cells under adherent and suspended conditions and how this difference affects cell apoptosis 3.Study the apoptosis of gastric cancer cells in adherent and suspended state;4.Study the effect of TF knockdown on the invasion and metastasis of gastric cancer cells,and then confirm the relationship between TF and anoikis;5.If TF has an impact on anoikis,explore possible mechanisms and pathways.Methods1.WB detects GES in normal gastric epithelial cell and gastric cancer cells MGC-803 SGC-7901,BGC-823,MKN45,and AGS with different degrees of differentiation,and selects TF-expressing cell lines;2.Knockdown the highest expression TF gastric cancer cell line,WB verifying the efficiency;3 Suspending 24 h and 48 h knockdown and non-knockdown of selected gastric cancer cells,WB verifying changes in TF expression at 24 h and 48 h respectively;4.Compared knockdown and non-knockdown of TF gastric cancer cell.Transewll,scratch test,CCK8,flow cytometry,etc.were used to detect cell invasion,apoptosis,migration and other characteristics;4.WB was used to verify whether TF was dehydrogenated through glycolysis and oxidative phosphorylation in mitochondria.Enzymes(LDH)and pyruvate dehydrogenase kinase(PDK)inhibit anoikis;5.WB verifying the expression of several antibodies in the classical apoptotic pathways;6.Compared the tumorigenic rate of gastric cancer cells in mice.Results1.WB results showed that compared with normal gastric epithelial cells GES,the expression of TF was highest in BGC-823;2.The experiment,CCK8,compared with the results of flow cytometry,compared with gastric cancer cells without TF knockdown,suggesting knockdown of gastric cancer cell invasion,metastasis,proliferation,and apoptosis were not significantly changed;3.Simulated gastric cancer suspensed,When simulated gastric cancer was suspensed,the results suggested that the expression of TF in both knockdown and non-knockdown cells all increased;4.Gastric cancer cells which knockdown TF and non-knockdown TF in suspension showed that the apoptotic rate of knockdown was higher than that of non-knockdown,indicating TF may be related to anoikis;5.Tumor formation experiments in mice suggested that gastric cancer cell of knockdowned TF was significantly smaller than non-knockdown TF.ConclusionIn summary,our results suggest that TF gastric cancer cells in invade,migrate,and proliferate have important influences,and promote their metastasis by inhibiting the anoikis phenomenon.This may be related to the energy metabolism pathway.PDK1 / 3 shows a difference.Apoptosis does not pass through the classic apoptotic pathway.At the same time,the tumor formation of knockdown TF was obvious.Therefore,TF may become a possible marker or target for gastric cancer diagnosis,which also provides a new idea for the diagnosis and treatment of gastric cancer.