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The Effect of G2/M Checkpoint Mediated by Cdc25c on Low-does Hyper-radiosensitivity in Human Lung Cancer Cell Line A549

Author: CuiYingShan
Tutor: ChengJing
School: Huazhong University of Science and Technology
Course: Oncology
Keywords: G2/M checkpoint low dose hype-radiorsensitivity Cdc25c
CLC: R730.5
Type: Master's thesis
Year: 2011
Downloads: 12
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Abstract


Objective To control the cell cycle checkpoint by down regulation of Cdc25c expression and observe the effect of cell cycle checkpoint on HRS/IRR,and discuss the mechanisms of HRS.Methods Hairpin siRNA templates targeting Cdc25c gene were synthesized and cloned into plasmid vector pGCsi-RAN-U6neo,transfected plasmids into human lung cancer cell line A549,selected the most effective plasmid by RT-PCR and Western blot, which were used to detect the expression of Cdc25c in the level of mRNA and protein. The cell lines including A549# transfected with pGCsi-RAN1 and A549* transfected with pGCsi-RANNC were constructed and irradiated with X-rays at different doses. Cell survival fraction was measured by the colony-forming assay based on ?uorescence-activated cell sorter, and the cell cycle distribution was examined with flow cytometry, and the expression of Cdc25c and P- Cdc25c(ser216) was detected by Western blot.Result The most effective plasmid is pGCsi-RAN1,and the expression of Cdc25c in A549# group was significantly lower than that in A549* group. HRS/IRR was observed in both cell lines while a exaggerated HRS was found in A549# cells. The ratio of G2/M phase cells was increased after exposure to X-ray doses >0.2Gy,while the cell cycle almost had no change at doses of <0.2Gy..A549# cells involved higher ratio of G2/M phase than that in A549* cells in unirradiated group, and which exhibited little difference at doses of <0.3Gy.After exposure to doses >0.3Gy,the ratio of G2/M phase cells was significantly increased and appeared ahead of time. These data indicate that high dose irradiation resulted in G2/M arrest and the threshold dose is different in these two kinds of cells. P- Cdc25c(ser216) protein expression was appeared and Cdc25c protein expression was decreased at doses of 0.3Gy in A549* cells while that is observed at doses of >0.3Gy in A549# cells. These data indicate that the expression of P- Cdc25c(ser216) and Cdc25c was accordance with G2/M checkpoint.Conclusions Low-does hyper-radiosensitivity may be associated with G2/M checkpoint which is mediated by Cdc25c and P- Cdc25c(ser216) protein.

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CLC: > Medicine, health > Oncology > General issues > Tumor Therapy
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