GDF11 Impact On The Growth And Metastasis Of A549 And H1299 Cells

Objective:Explore the relationship between GDF11 content in serum of patients with lung cancer and the patients of pathological features;Build PEGFP-GDF11 expression vector and explore the GDF11 exogenous expression of A549 and H1299 cell influence on growth and metastasis;Explore molecular mechanisms of A549 which high expression GDF11 impact on it cell growth and invasion.Method:1.The serum samples of patients with lung cancer were collected and the content of GDF11 in serum was detected by method ELISA.2.Cell A549 and H1299 were cultured in 15% serum of patients with lung cancer,and the changes of OD value after 24,48 and 72 h were detected by method CCK8.3.Design GDF11 primers,build PEGFP-GDF11 expression vector and verify4.Effects of up-regulated GDF11 expression on the growth and clone formation of cell A549 and H1299 were studied.5.Effects of up-regulated GDF11 expression on the transfer ability of cell A549 and H1299 by scratch experiment in vitro.6.Effects of up-regulated GDF11 expression on GLUT1,P53,MMP-2,the Bcl-2,Erk1/2 protein expression of A549 cells by western blotResults:1.The results of ELISA showed that the content of GDF11 in serum of patients with lung cancer was not related to sex(F=1.855,P>0.05)and lymph node metastasis(F=0.050,P>0.05).The content of GDF11 in serum was significantly related to the differentiation degree of cancer(F=24.567,P<0.05)and the age(F=3.980,P<0.05)of patients.2.The results of Cell A549 and H1299 were cultured in 15% serum of patients with lung cancer,in the serum group with high content of GDF11,the growth of cell A549 was significantly inhibited after 48h(F=3.975,P<0.05)and 72h(F=11.900 P<0.05),but the growth of cell H1299 was not affected.3.A549 and H1299 cell transfection PEGFP-GDF11,after testing the expression of its GDF11,visible results except GDF11 background 45 kd molecular protein expression,in about 70 kd have a more bright stripe,this suggests that the expression vector PEGFP-GDF11 successful heterologous expression in A549 and H1299 cells.4.Exogenous expression GDF11 within 24(t = 8.3,P < 8.3),48 h(t = 4.857,P < 0.05),significantly inhibit the growth of A549 cells and H1299 impact is not big.Clone forming experiments have shown that exogenous expression GDF11 can inhibit the A549 cell clone formation,but does not significantly influence on H1299 cells.5.Scratch the experimental results show that the GDF11 can inhibition invasion of A549 and H1299 cells.6.expression of exogenous GDF11 on A549 cell can inhibit the expression of GLUT1,Bcl-2,MMP-2 and Erk1/2 protein;Exogenous expression GDF11 on A549 cells can promote the expression of P53 proteinConclusion:1.GDF11 of content in serum of patients with lung cancer is related to its pathology,high malignant degree of poorly differentiated GDF11 of content in serum of patients with lung cancer is low.Secondly,the content of serum GDF11 associated with patient age,the age and the smaller the patients serum GDF11 content is higher2.Exogenous expression GDF11 can inhibit the growth of A549 cells and the formation of the clone,but for H1299 cells was not significant;Exogenous expression GDF11 significant inhibition of A549 and H1299 cells transfer ability3.GDF11 regulating the expression of Glut1 by activation of P53;and reduce the expression of apoptosis related protein Bcl-2;regulation of expression MMP-2;Thus inhibit the growth and metastasis of the cell.