Effect Of Apoptosis Of Transfection TRA2B SiRNA On Lung Adenocarcinoma A549 Cells And Clinical Analysis Of Lung Cancer In Elderly Patients

Objective:TRA2B is the member of rich in serine / arginine-like protein family,which involved in pre-mRNA alternative splicing.The study find that TRA2 B highly express in cervical cancer, lung cancer, breast cancer, ovarian cancer, stomach cancer, colon cancer and other cancers. Nowdays there is already designed to TRA2 B targeting cancer gene therapy, and obtained preliminary results. But researches on TRA2 B gene expression in lung cancer is still rare at home and abroad. In this research,after we transfect siRNA TRA2 B into the A549 cell by RNA interference technology,we observe morphological changes in A549 cells, TRA2 B expression in A549 cells, and the effect of apoptosis of A549 cells. We explore the mechanisms about TRA2 B occurring in lung cancer, and provide a theoretical basis for further TRA2 B as targeted therapy in lung cancer gene targets. By analysis of the elderly lung cancer cases, discuss the significance imaging in the early diagnosis and early treatment of lung cancer.Methods:We artificially construct siRNA-TRA2 B for the TRA2 B gene in lung adenocarcinoma. Through the use of liposomes Lipofectamine TM 2000, the siRNA-TRA2 B successfully transfect into A549 cells, and observe morphological changes under a microscope. We detect the TRA2 B mRNA expression levels by Realtime PCR, and filter out the most effective interfering RNA. We detect the TRA2 B protein expression levels by Western blot,and filter out the most effective interfering RNA. After transfect the best interference effect siRNA-TRA2 B, we use flow cytometry to detect the level of apoptosis of A549 cells. And we use Western blot to detect the apoptotic protein Bcl-2 and Bax protein expression levels. We discuss the effect ofapoptotic protein Bcl-2 and Bax expression after transfection.By selecting three cases of lung cancer in elderly patients are analyzed, compared with the postoperative outcome of cases, investigate the clinical significance of radiographic examination of lung cancer found early.Results:Through the use of liposomes LipofectamineTM 2000, the siRNA-TRA2 B successfully transfected into A549 cells:(1) Observed under an inverted microscope before and after A549 cells were transfected cell morphology: before transfection, cells were in good condition, adherent growth, mostly fusiform cytoplasm plump, medium size, clear nucleoli, transfection after 24, 48 hours, cell wrinkle shrink, adherent poor,transfection efficiency of over 90 %.(2) Realtime PCR detection TRA2 B mRNA expression level, and select the best siRNA interference effect, the result shows: the siRNA- TRA2 B effect on A549 cells after 24 h, 48 h, TRA2 B mRNA expression significantly lowered, and the Mock group, Negative Control group and the Black group of the mRNA expression level has no obvious change, at the same time, select 1-TRA2B- homo- 544 group interference effect is best.(3) Western blot detection TRA2 B protein expression level, and selected the best si RNA interference effect, the results showed: siRNA- TRA2 B effect on A549 cells after 24 h, 48 h, TRA2 B protein expression significantly decreased, while the Mock group, Negative Control group and the Black group of the protein expression level has no obvious change, at the same time,select 1- TRA2B- homo- 544 group interference effect is best.(4)Flow cytometry detection apoptosis of A549 cell level, after transfection interference effect best siRNA-TRA2 B, the result shows: the apoptosis rate of siRNA- TRA2 B group(48.02±0.495) %,Negative Control group(8.60±0.251) %, Mock group(8.47±0.156) %,Black group(9.10±0.355) %, siRNA TRA2 B compared with Mock group, Negative Control group and the Black, the difference was statistically significant(P < 0.05), the Mock group,Negative Control group compared with between two Black group, there was no statistically significant difference(P > 0.05). It can promote cell apoptosis after transfection siRNA- TRA2 B.(5) Western blot detection the apoptotic protein Bcl-2 and Bax protein expression levels, after transfection interference effect best siRNA-TRA2 B, the result shows: in A549 cells, the apoptosis protein Bcl- 2 protein expression level decreased obviously, the Bax protein expression level increased significantly,while the Mock group, Negative Control group and the Black group of the Bcl- 2, Bax protein expression level has no obvious change. 3 cases of imaging results consistent with the pathological results, imaging can be used as an important means of early detection of lung cancer.Conclusion:By using RNA interference technology, down TRA2 B expression in A549 cells, inhibition of cell proliferation and promote apoptosis, TRA2 B will likely become a new target for cancer gene therapy. Imaging of early lung cancer in elderly patients found that early diagnosis and early treatment is of great significance.