The Comparative Research Of The Change Of Residual Proliferation,Migration,and Stem Cells Of Residual Cells,the Cells In Which Are The Two Lung Adenocarcinoma Cells After Radiation In Vitro

Purpose: By adding different doses of X-ray irradiation to Xuanwei lung adenocarcinoma XWLC-05 and normal lung adenocarcinoma A549 cells,then through recording and analyzing the changes of their two cell proliferation, migration and cancer stem cells,the study will compare the differences between them in the?biological behavior of the remaining cells and relevant mechanism. This study aims to guide the radiation and comprehensive treatment of lung cancer.Materials and methods: A549 and XWLC-05 cells exposure to 0,4 and 8Gy with 6MV x-ray. The control group (OGy) did not receive radiation, and the other treatments were the same as the radiation group. We used the Colony Formation Assayto detect the surive of the cells after radiation. Cell proliferation assay to detect the proliferation of the cells after radiation, Cell migration assay to detect the migration of the cells after radiation, Flow cytometry was used to analyze the changes of cell cycle,apoptosis and tumor stem cells after radiation,and Western Blot was used to analyze the changes of PCNA,MMP2,P53. To study the changes and differences of biological behavior of A549 and lung adenocarcinoma cell line XWLC-05 after radiationResults :? The radiosensitivity of XWLC-05 and A549 was detected by clone formation assay(figurel).With the increasing of radiated dose, SF was decreased gradually , the survival fraction of A549 cells was higher than that of XWLC-05 cells . The results indicated that the radiosensitivity of XWLC-05 cells was higher than that of A549 cells.? Within 96h after radiation,cell viability of A549 and XWLC-05 irradiated group was suppressed, whereas, 360h after radiation, the activity of XWLC-05 irradiation group was enhanced .The cell viability of A549 was also weakened than control group.The activity of XWLC-05 cells was stronger than that of A549 cells after radiation.?The migration ability of A549 and XWLC-05 cells was changed after radiation in a time dependent manner. 24-96 h after radiation, the migration ability of A549 and XWLC-05 cells was significant weakened than control groups, but 336h after radiation , the migration ability of the radiation group was enhanced than control groups .336h after radiation, compared with control group, the migration ability of two cells was enhanced, and the migration ability of XWLC-05 cells was stronger than that of A549 cells. 336h after radiation, the migration ability of XWLC-05and A549 cells at a dose of 8 Gy was significantly weakened than 4Gy radiation.?The cell cycle of A549 and XWLC-05cells were changed in different degree after irradiation . (1)TheGO/ G1 phase in A549 cells was arrested after radiation .Compared with unirradiated control group , the G0/G1 phase of the XWLC-05 radiated group was decreased,attaining a minimum at 12h after radiation. The proportion of GO/ G1 phase in 8Gy irradiated group was lower than that of 4Gy irradiated group. (2)Compared. with unirradiated control group, the proportion of G2/M phase of the irradiated A549 and XWLC-05 cells were increased after radiation.The proportion of G2/M phase in 4Gy irradiated group was lower than that of 8Gy irradiated group. (3)The proportion of S phase of the radiated A549 cells did not exceed that of the unradiated A549 cells 12-to 192-h after radiation .Compared with unirradiated control group , the proportion of S phase in irradiated XWLC-05 groups was temporary increased 12h after radiation and dropped below the unirradiated control level at 48h after radiation ,then begun to exceed the unirradiated control level.?CD133 and CD44 expression on the surface of cancer stem cells in A549 cells was not significantly changed after irradiation.However, Compared with unirradiated XWLC-05 cells ,CD44 and CD 133 expressions in XWLC-05 cells was increased significantly.? Compared with unirradiated control group,the cell apoptosis rate of the two irradiated groups(irradiated A549 group and irradiated XWLC-05 group )were increased after radiation, and both of them in 8Gy radiation was higher than that of 4Gy radiation. The highest cell apoptosis rate in A549cells was seen 192h after radiation, p<0.05. The highest cell apoptosis rate in XWLC-05cells was seen 48h after radiation, p<0.05.? The MMP-2 and PCNA expressions was not consistent with cell migration and proliferation. But the expression of P53 was enhanced ,and was consistent with the cell apoptosis of A549 and XWLC-05 cells .Conclusion: It is different in the changes of proliferation and migration between XWLC-05 cells and A549 cells after radiation. Long-term proliferation and migration in XWLC-05 cells was enhanced after radiation . Long-term migration was also enhanced, but proliferation was suppressed in A549 cells after radiation.Proliferation and migration in XWLC-05 cells are stronger than that in A549 cells.The migration ability of XWLC-05 and A549 cells at a dose of 8 Gy wassignificantly weakened than 4Gy radiation. A549 cells after CD 133 irradiation, the expression of CD44 did not change significantly, and the residual XWLC-05 cell CD 133, CD44 expression increased significantly. MMP-2, the main factors of PCNA may not be involved in the radiation induced cell proliferation and migration, and the expression of P53 after radiation and apoptosis are closely related.