Mechanism Of Serum Starvation-induced Migration And Invasion Of Lung Cancer Cells NCI-H1650

Cancer is one of the major diseases that endanger human health.It is mainly characterized by abnormal growth of cancer cells,which can be transferred from the primary site to other tissues or organs of the body,and eventually form metastatic cancer.Epithelial-Mesenchymal Transition(EMT)is one of the main ways of cancer metastasis.In addition,changes in the microenvironment and mediation of MAPK signaling pathways can also promote cancer metastasis.However,the role and mechanism of tumor cell invasion and migration are still unclear when tumor cells are under low serum secretion factors.In order to investigate the effect of serum withdrawal on tumor cell morphology,migration ability and invasion ability,this experiment conducted serum starvation experiments on non-small cell lung cancer NCI-H1650 cells.The results showed that after serum starvation,NCI-H1650 cells changed from the initial epithelial to fibroblastic morphology,and the cell motility,transwell migration ability and invasion ability were significantly enhanced.Furthermore,after serum starvation treatment of NCI-H1650 cells,the final concentration of 2% FBS(Fetal Bovine Serum)was added,It was found that fibroblast-like cells returned to epithelial-like after starvation,and the migration ability and invasive ability were obvious decline.In addition,in order to investigate whether the migration ability and invasion ability of NCI-H1650 cells caused by serum starvation treatment are related to EMT,this experiment verifies the expression level of EMT-related markers from protein level and mRNA level(Vimentin,Snail1,E-Cadherin).The results showed that the expression level of transcription factor Snail1 was lower in cells with 2% FBS,but the expression level was significantly up-regulated in serum-starved cells.The expression of Snail1 was significantly down-regulated after 2% FBS recovery.In order to further clarify whether Snail1 plays an important role in the process of serum starvation-induced NCI-H1650 cell migration invasiveness,this experiment performed siRNA interference assay on NCI-H1650 cells,and silenced Snail1 gene before serum starvation treatment.The results showed that Compared with the control,there was no significant difference in morphology and migration ability after Snail1 knockdown under conditions of 2% FBS or complete serum withdrawal.It is concluded that the migration ability and invasion ability induced by hunger stimulation are not related to EMT.In order to find the key regulators that regulate serum starvation and promote the invasion of NCI-H1650 cells,this study identified stress-signal-related channels,such as MAPK,and verified the pharmacological experiments with pathway inhibitors.Finally,under serum starvation conditions,ERK protein phosphorylation levels are significantly elevated,resulting in changes in cell morphology and promoting cell invasion and migration.