| Objective:we choose pheiffer cell line which has similar biological characteristics with the orbital diffuse large B-cell lymphoma as our experimental subject.To observe the effects of diffetent dosage of X-ray and different time on proliferation,cell cycle and cell apoptosis of the diffuse large B-cell lymphoma (DLBCL) cells.With the result of this study,we can research the effect of X-ray on DLBCL cells which can afford some new ideas for its comprehensive therapy.Methods:1. After anabiosis, the proliferative status and morphological characteristic of DLBCL cells were observed.2. DLBCL cells were plated in 96-well plates and cultured in medium with 10% fetal bovine serum for 24h. Then they were divided into control group (0 Gy) and test groups (2Gy,4Gy,6Gy,8Gy) at random. After radiation with X-ray, the cells were cultured for 6h,12h,24h,48h,72h,96h respectively.In the process, the morphological change was observed, cell proliferation was determined by MTS/pms and survival fraction(SF) was calculated.3. Analyze the effect of different dosage and different time on the cell cycle and cell apoptosis of DLBCL cell by FCM.Results:1. DLBCL cells developed well and passed stably. The shape of cell was round or ellipse.2. The optical density(OD) of test groups were lower than control group.In the same clutrued time,the higher dosage was,the lower OD was. When 8Gy group's cell was cultured for 6h,6Gy group's cell was cultured for 24h,4Gy group's cell was cultured for 48h,2Gy group's cell was cultured for 72h,the OD were significantly lower than control group(P<0.05). Cell growth was slower when the dosage increased.In the meantime,SF declined gradually with the dose increasing and the time expandsion.3. At 6h after irradiation,the number of G2-M stage cells of each test group was slightly low (<17%).However, at 12h after irradiation,the number of test group was increased gradually which was significantly different from the number of control group(P<0.01).The proportion of G2-M stage cells of 8Gy group raised gradually,even not recoverd at 48h.While the proportion of 2Gy,4Gy and 6Gy groups rised from 6h to 24h,then it declined slowly..4,The apoptotic rate of tumor cells augmented gradually with the dosage inc-reasing.In the same treating time,the rate of test groups was increased significantly in contrast with the rate of contral group(P<0.01).The apoptotic rate was higher when the cell of test groups was cultured for a longer time after irradiation.5,At 12h after irradiation, the apoptotic rate of 4Gy,6Gy and 8Gy groups are abviously high.It was increased with the time extension.Combine with the curve chart of cell cycle,significant blockage of G2-M stage also occurred at 12h after irradiation.It shows that apoptosis maybe come from G2-M stage.Conclusion:1. X-ray can inhibit the proliferation of DLBCL cell in dose-dependent, time-dependent manner.2. X-ray increase the proportion of the G2-M stage cells.With the dasage increasing,the number of G2-M stage cells is higher and the time of blockage is longer.3. The apoptotic rate of tumor cells augment gradually with the dosage increasing and time expansion.4. Apoptosis which is induced by X-radiation maybe come from G2-M stage.It is illustrated that delayed interphase apoptosis occurs in DLBCL cell.5. With the result of this study, we presume that fraetionated radiation and the drug which specially shorten G2-M stage may paly an important role in therapy of DLBCL.It also affords a new idea and direction for clinical treatment of DLBCL. |
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