The Study Of The Antitumous Effect Of RBM10 On Human Lung Adenocarcinoma Cells

Objective:RNA binding motif protein family(RBM) is a newly discovered protein family with Similar structure and function,which may be related to the splicing,translation and stability of m RNA.RBM10 and RBM5 are important members of the RBM.The present study showed that RBM10 and RBM5 with similar structure and high degree of homology are potential tumor suppressor gene. The existing evidence suggested that RBM5 is a tumor suppressor gene in lung adenocarcinoma,which can inhibit the growth and proliferation of lung adenocarcinoma cells. However,whether RBM10 could inhibit the growth and proliferation of lung adenocarcinoma cells is still unknown. In order to explore the role of RBM10 in the mechanism of pathogenesis of lung adenocarcinoma,we collected 41 cases of patients with primary lung adenocarcinoma of lung cancer and normal lung tissue samples(taken from the same patient),respectively.Reverse transcription-polymera chain reaction(RT-PCR)and Western blot were used to observe the expession of RBM10 m RNA and protein in lung adenocarcinoma,and correlation between the expression level and clinical pathological parameters of lung adenocarcinoma was further analyzed. Additional,A549 and H1299 cells infected with recombinant lentivirus were further observed in vitro to study the effect of stable RBM10 overexpression on the proliferation and cell cycle of lung cancer cells in order to explore the molecular mechanism of the role of RBM10 in the pathogenesis in lung adenocarcinoma,which can lay the foundation for the explore the novel target gene and advanced molecular targeted therapies in lung cancer.Methods:1. RT-PCR and Western blot were used to detect expression of RBM10 m RNA and protein in 41 cases of human lung cancer tissue,and to analysis the correlation between RBM10 protein expression and clinicopathological parameters(including sex,age,smoking history,lymph node metastasis,lung TMN staging) on this basis.2.A549 and H1299 cells was infected with the recombinant lentivirus in order to obtain the cell lines with stable overexpression of RBM10 successfully.3.Proliferation assay(MTT) of A549 and H1299 cell lines induced by stable RBM10 overpression.4.Colony formation of A549 and H1299 cell lines induced by stable RBM10 overpression.5.Flow cytometry method was used to observe the effect of the cell growth cycle in A549 and H1299 cell lines induced by stable RBM10 overpression. Result:(1) Compared with the adjacent tissues,RBM10 m RNA and protein expression in lung adenocarcinoma tissues were significantly reduced,even loss,the difference was statistically meaningful(P<0.05).There is on correlation between expression level of RBM10 protein and clinicopathological parameters(including sex,age,smoking history,lymph node metastasis) in human lung adenocarcinoma tissues,but is negatively correlated with TNM stage(P<0.05).(2)We successfully abtain the A549 and H1299 cell lines with stable overexpression of RBM10.(3)MTT experimental results showed that stable overexpression of RBM10 significantly inhibited the proliferation of A549 and H1299 cell lines.(4)Cloning experiment showed that the stable RBM10 overexpression significantly inhibited the cell clone ability in A549 and H1299 cell lines.(5) Cell cycle experiments showed that the stable RBM10 overexpression can block the A549 cell cycle in the G0/G1 phase,while the H1299 cell cycle arrest in the S phase. Conclusion:(1)The expression of RBM10 in human lung adenocarcinoma was down regulated,and which was negatively correlated with the TNM stage,suggesting that RBM10 might be involved in the occurrence and development of lung adenocarcinoma.(2)Stable RBM10 overexpression inhibits the growth of A549 cells and H1299 cells,reduced cell clonogenic capacity and induced cell cycle arrest. The results suggest that RBM10 expression might play an important role in lung adenocarcinoma.