The Study Of Effect Of Amifostine On The Proliferation Of Hela Cell With Radiotherapy

ObjectiveCervical cancer is one of the most common malingnancy in women and seriously affecting their health. Radiation treatment is an important therapeutic option, combined with surgery and chemotherapy can get long-term survival. However the treatment is frequently limited by adverse effects on normal tissues. Amifostine as a broad-spectrum cytoprotective agent its selective normal tissue protective mechanism include free-radical scavenging, DNA and cell membranes protecting. The U.S.Food and Drug Administration has approved the i.v. use of amifostine to reduce the cumulative renal toxicity of cisplatin and to reduce the incidence of moderate to severe xerostomia in patients undergoing radiotherapy for head and neck cancer. Amifostine is not recommended to use in cervical cancer treatment because the protective effect can not be ruled out.The objective of this in vitro experiment is to observe the cell kinetics effects of amifostine on irradiated Hela cells as a radiation protector, to provide scientific theory for the ppplication of amifostine in cervical cancer treatment, find new ideas and methods on cervical cancer treatment.MethodsAmifostine was observed using MTT method on Hela cell proliferation rate. Using cell colony-forming experiment to observe the radiation protective influence of amifostine to cervical cancer Hela cells. Using FCM to analysis of 6MV-X ray irradiation alone and irradiation combined AMI on Hela cell apoptosis and cell cycle. Random set radiotherapy alone group (R) and radiation+drug group (D+R) to carry out cell colony-forming experiment and drawing survival curves. Random set control group (C),drug group (D),radiotherapy alone group (R) and radiation+drug group (D+R) were treated with flow cytometry cell apoptosis and cell cycle in each group.Results1.MTT assay shows there is no difference between each group with AMI(25,50,100,200,400,600 ug/ml) act on Hela cells after 24 hours and 48 hours, cell proliferation is not inhibited to varying degrees, showing no significant dose-effect relationship.2.Using the experimental results with colony-forming drawn survival curves and related parameters show that all parameters of radiation+ drug group(D+R) are corresponding smaller than the radiotherapy alone group (R), the Do of the radiotherapy alone group (R) were less than radiation+drug group(D+R).Tt is shown that after adding AMI the radiation sensitivity of Hela cells was increased, and the reducing of the value of Dq showed amifostine can be degraded Hela cells to sublethal radiation damage repair capacity.3. Flow cytometer FCM showed amifostine lead to G1 arrest in 24h and G2 arrest in 72h. FCM in each group Hela cells apoptosis,the results showed that:single-drug group was 3.15%,3.36%. Radiotherapy alone group:5.10%,5.43%,6.89%,7.21%. Drug+irradiation group: 25μg/ml to 5.41%,6.25%,9.82%,10.54%; 100μg/ml to 7.25%,7.98%, 10.86%,11.41%.Drug+radiation group (D+R) increased more apoptosis than radiotherapy alone (P> 0.05).Conclusion:1.There was no significant protective effet of AMI to Hela cells.2.Amifostine can increase the radiosenditiviity of cervical cancer Hela cells,its molecular mechanism may be amifostine can lead to cervical Hela cells G2 arrest and promote apoptosis.3Amifostine can be used as an effective cytoprotective agent in cervical cancer radiotherapy.