癌相关成纤维细胞对喉癌细胞系Hep-2生物学行为影响的体外研究

目的 探讨癌相关成纤维细胞(CAFs)在体外的分泌功能及对喉癌细胞Hep-2生物学行为的影响.方法 提取江西省肿瘤医院2016-09-09 1例喉鳞状细胞癌手术患者的癌组织及癌旁组织,进行喉癌CAFs及正常成纤维细胞(NFs)的原代培养.ELISA试剂盒测定CAFs和NFs培养上清液中MMP-2、MMP-9的浓度.制备...

Effects of CAFs on biological behaviors of laryngeal carcinoma cell line hep-2 in vitro

Objective To investigate the secretory function of carcinoma associated fibroblasts(CAFs)and the effects of CAFs on biological behavior of laryngeal carcinoma cell line Hep-2 in vitro.Methods Tumor and adjacent histologically normal tissues of patient with laryngeal squamouos cell carcinoma from Department of Head and Neck Surgery,Jiangxi Cancer Hospital in September 9,2016 were collected for primary culture of CAFs and normal fibroblasts(NFs).The concentrations of MMP-2,MMP-9 in NFs and CAFs cell supernatant were measured by ELISA.Hep-2 cells were cultured with CAFs and NFs conditioned medium respectively,and the cell proliferation was determined by CCK-8 assays.After co-cultured with CAFs and NFs respectively,the apoptosis and invasion of Hep-2 cells were evaluated by flow cytometry and Transwell assay.The experiment was divided into 3 groups:experiment group(co-cultured with CAFs),control group(co-cultured with NFs)and empty group(cultured with DMEM).Results The levels of MMP-2 in CAFs cell supernatant and NFs cell supernatant were(3.40±0.22)ng/ml and(2.52±0.19)ng/ml respectively.The difference was statistically significant(t=-5.246,P=0.006).The difference between the levels of MMP-9 in CAFs cell supernatant and NFs cell supernatant was also statistically significant with the concentrations of(164.88±6.54)pg/ml vs(114.16±4.48)pg/ml(t=-11.087,P<0.001).CCK-8 results through statistical analysis suggested that CAFs conditioned medium could significantly promote the proliferation of Hep-2 cells compared with the NFs condioned medium(F=13.977,P<0.001).The apoptosis rate of Hep-2 cells in experiment group was(7.17±1.24)%,while in control group and empty group were(2.45±0.73)%and(2.05±0.74)%respectively,the difference was statistically significant(F=27.664,P=0.001).The number of Hep-2 cells that penetrated the upper chamber layer in experiment group was 347.80±23.73,while the control group and empty group were 254.00±46.11 and 191.40±23.67,respectively.The difference was statistically significant(F=28.599,P<0.001).Conclusions CAFs promote the proliferation,invasion and induce apoptosis of Hep-2 cells.The upregulation of MMP-2,MMP-9 secretory expression in CAFs may contribute to enhance the invasiveness of Hep-2 cells.