Effect Of The Apoptosis Of Human Adenocarcinoma Of The Uterine Cervix Hela Cells Induced By 5-Fluorouracil(Fu) Combined With Radiotherapy(RT)

Uterine cervix cancer is one of the frequent maligant tumor in the department of gynecology.Now,opertion and radiotherapy are still the major traditional therapeutic methods for patients with cervical cancer.By the two methods,some patients recover,some,however,themajority of them become worse.Nowadays,adenocarcinoma of the uterine cervix has a gradually increased tendency,which is easily recur or transfer treated by operation alone or RT alone in a short time,it is also less sensitive to chemotherapy(ChT) alone,so it has a worse prognosis.In order to improve prognosis,it is urgent to find out a better way to cure adenocarcinoma of the uterine cervix.Nowdays it is reported that some new therapeutic tools such as neoadjuvant chemotherapy(chemotherapy before surgery or radiotherapy),concurrent chemotherapy and radiotherapy etc.Which could be employed to shrink the focal volume of the cervix cancer,to eliminate effectively the inferior clinical transfer,to improve the effect of operation or radiation.So,now the chemotherapy combined with radiotherapy had became a study point in tumor-treated aspect.Now 5-Fu is usually used to treat many maligant,and one of the neoadjuvant chemotherapy medicines of adenocarcinoma of the uterine cervix,5-Fu also had invariably radiosensitization.This text to begin with apoptosis angle,approached the mechanism of adenocarcinoma cerical cancer Hela cell induced by 5-Fluorouracil(Fu) combined with radiation.To investigate the effect of the apoptosis of Human cervical cancer Hela cells induced by 5-Fluorouracil(Fu) combined with radiotherapy(RT).The MTT assay,flow cytometry,fluorescent microscopy were used.Materials and MethodsMaterial Cell Strain Hela cell strain of human cervical adencarcinoma from laboratory of infection department in Shengjing Hospital affiliated to China Medical University,which are cultured in 1640 culture mediumand 10%bovine serum albumin.Reagent Text box for Annexin V/PI buy from boaosen beijingExperiment MethodsHela cells are cultured in 1640 culture mediumand 10%bovine serum albumin. The non-cytotoxic concentration(48 h IC₅₀)of Hela cells treated with 5-Fu was determined by methyl thiazolyl tetrazolium(MTT).The logarithmically growing Hela cells were plated at a density of 1×10⁶cells/ml,a total of 2 ml were seeded on each well of a 6-well plate and incubated for 24 h.The Hela cells were divided randomly into control group(the group have no treatment),chemotherapy(ChT) alone(After obtaining 48 h IC₅₀ of 5-Fu on Hela cells through MTT,We use the 5-Fu of higher than or tantamount to 48 h IC₅₀),RT alone(2,4,6 Gy),RT combined with ChT group(first treated by 2,4,6 Gy RT and than treated by different doses 5-Fu).The cells after treatment incubated for 24 h,48 h,then the cells were centrifuged and collectd.Apoptosis was quantified by using Annexin V-FITC/PI(propidium iodide),the cells were measured with FITC/PI staining using flow cytometry.Microscopic observations were performed and photographed using a fluorescence microscope.Statistical AnalysisExperimentmal data has been statisticed by SPSS 13.0.Results were expressed as mean values±standard deviation.Statistical significance of apoptosis group was analyzed by two-tailed Student's t-test.Statistical significance of apoptosis interclass was analyzed by One-way ANOVA.P＜0.05 was considered to be significant.P＜0.01 was considered to be statistically significant.ResultsThe 50%inhibitory concentration(IC₅₀) value of Hela cell treatment by 5-Fu for 48 hours is 125μg/ml through MTT assay.ChT alone,RT alone and RT combined with ChT could induce the apoptosis of Hela cells,RT combined with ChT could obviously enhance the apoptosis of Hela cells.The apoptosis cells can be detected by using a fluorescence microscope. Conclusions5-Fu combined with radiotherapy(RT) can obviously enhance the apoptosis of Hela cells.The study provides the academic proof for clinical use of 5-Fu combined with radiotherapy(RT) can enhancing Hela cell apoptosis.