Influence Of Body Fluid Of Ascaris Lumbricoides On The Secretion Of Interleukin-6 And Transforming Growth Factor β Of Lung Adenocarcinoma Cell Line (A549)

Objective:To study the influence of body fluid of Ascaris lumbricoides on the secretion of interleukin-6 and transforming growth factor β of lung adenocarcinoma cell line (A549) in the following two aspects: firstly, the relationship between the ABF concentration and inducing time with the secretion level of IL-6 and TGF-β ,and secondly, the relationship between the level of the two cytokines with the apoptosis of A549 induced by ABF. Methods:1,Obtainment of growth curve of A549 cell: Cells in the logarithmic phase were digested and counted, then inoculated into 24-well cultivation plate. Cell counts in three holes were made every day for following 7 days to observe the growth dynamic of the cell growth and determine the proper time for next experiments.2, ABF collection: Intact Ascaris lumbricoides adult worms were collected from children after an oral administration of pyrantel pamoate at one dosage of 30 mg/kg of body weight. Worms were washed separately with tap water and sterile saline. After drying with tissue paper, the worm was cut at the end of the tail to collect ABF. The fluid was spun at 10,000 ×g for 30 minutes to remove particles, then sterilized by a 0.45μm filter and stored at -30℃ for later use.3,Cytotoxicity test: Cells in the logarithmic phase were counted and inoculated into 96-well cultivation plate exactly. ABF with initial concentration of 3370μg/ml was diluted mutltiproportionly into 13.5μg/ml. The method of MTT (3-(4,5-dimethyl-thiazol-2-yl)-2,5- diphenyltetrazolium bromide) was used to test...