The Study Of The Radiotherapy Sensitization Effect Of TMP For ECA109 Cell

Objective: This study mainly research on the effect of TMP,radiotherapy and both on proliferation and apoptosis of ECA109 cell to provide theoretical basis for cancer treatment.Methods: CCK-8 method was used to detect the inhibition rate of TMP,radioactive rays single acting and combined them application on the ECA109 cell.Observed the morphologic change of cells treated with TMP,radioactive rays single acting and combined them application on ECA109 cell.Flow Cytometry was used for the detecting apoptosis rate and cell cycle.Double immunofluorescence staining is exploited to detect the apoptosis of ECA109 cells treated with TMP and radioactive rays.Apoptosis was measured using DNA fragmentation.Western-blot technique was used to detecte the expression of Bax and Bcl-2 protein after the ECA109 cells were treated with TMP and radioactive rays.Results: 1 TMP and radioactive rays could inhibit the proliferation of ECA109 cells in a dose dependent manner.Joint action is effective than single drug.2 Under the inverted microscope,we can see that the morphology of ECA109 cells changed in different degree after treated with TMP and radioactive rays.3 TMP can make cell cycle arrest in G0/G1 phase and induce apoptosis,radioactive rays can make cell cycle arrest in G2/ M phase.This effect was much more obvious when TMP and radioactive rays were combined together.4 Results of flow cytometry showed that the apoptosis rate of ECA109 cells treated with TMP,radiotherapy single acting and combined them application for 24 h was:(13.57±1.32)%,(9.14±2.01)%,(15.7±0.13)%,compared with the control group,has distinct difference(P<0.01).5 Agarose gel electrophoresis results showed that: Ladder's band was seen in the group B and group D.6 The protein of Bcl-2 was decreased,Bax was increased in TMP single acting group.Bcl-2 and Bax protein expression had no significant effect in the radioactive rays single acting group.Compare with TMP single acting group,Bcl-2 protein expression was much lower in TMP and radioactive rays combined group.Conclusions:1 TMP and radioactive rays both could inhibit ECA109 cells,the inhibition effect has dose dependency.Joint action is effective than single drug.2 The inhibition effect of TMP and radioactive rays is relevant to their function of inducing apoptosis.3 TMP have radiotherpy sensitization effect on ECA109 cells.4 The mechanism of TMP enhance radiosensitivity on ECA109 cells may be due to its regulation on cell cycle,induce apotosis and down-regulating the expression of bcl-2 protein.