Latexin对耐吉西他滨胰腺癌细胞化疗改善作用及其可能机制的体外研究  

In vitro study on the improvements of Latexin on the chemosensitivity in gemcitabine-resistant pancreatic cancer cells and potential mechanism

作　　者：郑继行[1] 王成[3] 周香[1] 薛战雄[2] 蔡振寨[2] 

机构地区：[1]温州医科大学附属第二医院普外科,浙江温州325000 [2]宁波市第一医院普外科 [3]温州医科大学附属第二医院消化内科,浙江温州325000

出　　处：《中华胰腺病杂志》2017年第2期109-113,共5页Chinese Journal of Pancreatology

基　　金：浙江省自然科学基金（Y2100546）

摘　　要：目的 观察Latexin干预对耐吉西他滨胰腺癌SW1990细胞耐药性的影响,探讨其可能机制.方法 采用吉西他滨药物浓度梯度递增诱导法建立吉西他滨耐药SW1990细胞株（SW1990/GZ）.采用CCK-8法检测吉西他滨对SW1990、SW1990/GZ细胞的IC50,并检测10、20、40 ng/μl Latexin干预细胞48 h后的IC50,计算耐药指数（RI）.采用定量RT-PCR 法、蛋白质印迹法检测SW1990、SW1990/GZ细胞Latexin基因mRNA和蛋白的表达,检测40 ng/μl Latexin干预48 h后SW1990、SW1990/GZ细胞Shh、Gli1的mRNA和蛋白表达.结果 成功建立了SW1990/GZ细胞株,它在含150 μmol/L吉西他滨的培养液中稳定生长、传代. 吉西他滨对SW1990、SW1990/GZ细胞的IC50值分别为（3.8±0.4）μmol/L和（226.5±13.6）μmol/L,SW1990/GZ细胞显著高于SW1990细胞,差异有统计学意义（P=0.000）;RI均值为59.6.10、20 、40 ng/μl Latexin干预48 h后,吉西他滨对SW1990细胞IC50值分别为（3.0±0.4）、（2.5±0.3）、（1.8±0.3）μmol/L,对SW1990/GZ细胞的IC50值分别为（113.08±5.01）、（70.26±2.31）、（42.12±1.31）μmol/L,除10 ng/μl Latexin干预的SW1990细胞外,其余剂量Latexin干预的SW1990、SW1990/GZ细胞IC50值均较未干预细胞显著下降,差异具有统计学意义（P值均〈0.05）;RI均值分别为37.7、28.1、23.1.SW1990、SW1990/GZ细胞Latexin的mRNA相对表达量分别为0.85±0.08、0.31±0.07, 蛋白相对表达量分别为0.49±0.09、0.13±0.05,SW1990/GZ细胞的表达量显著低于SW1990细胞,差异均有统计学意义（P值均〈0.01）.40 ng/μl Latexin干预48 h后,SW1990/GZ细胞Shh mRNA表达量从未干预细胞的0.89±0.09下降至0.53±0.06,Gli1 mRNA表达量从0.58±0.06下降至0.35±0.05;Shh蛋白表达量从0.72±0.09下降至0.35±0.06,Gli1蛋白表达量从0.78±0.08下降至0.28±0.03,差异均有统计学意义（P值均〈0.01）.结论 Latexin能增强耐吉西他滨胰腺癌SW1990细胞对吉西他滨的敏感性,其机制可能与抑制SHH通路的激活有关Objective To observe the effect of Latexin treatment on the chemoresistance in gemcitabine-resistant pancreatic cancer cell line SW1990, and explore the potential mechanism.Methods Gemcitabine-resistant pancreatic cancer cell line SWl990/GZ was induced and established by increasing gemcitabine dosage intermittently.IC50 of gemcitabine in SW1990 cells and SWl990/GZ cells pre and post Latexin treatment at the dosage of 10, 20 and 40 ng/μl for 48 h was evaluated using CCK-8 assay.The mRNA and protein expression of Latexin gene in SW1990 and SW1990/GZ cells were evaluated using qRT-PCR and Western blot, and the expression of Shh and Gli1 in 40 ng/μl Latexin treated SW1990 and SW1990/GZ cells for 48 h.Results A gemcitabine-resistant pancreatic cancer cell line SWl990/GZ was obtained successfully, which can grow stably and passage in the media containing 150 μmol/L gemcitabine.The IC50 values of gemcitabine in SW1990 cells and SWl990/GZ cells were （3.8±0.4）μmol/L and（226.52±13.61）μmol/L, respectively, and the later was greatly higher than the former, which was statistically different （P=0.000）.The drug resistance indexes （RI） was 59.6.After treated with different concentrations of Latexin（10,20,40 ng/μl）, the IC50 of SW1990 cells was （3.0±0.4）μmol/L, （2.5±0.3）μmol/L and （1.8±0.3）μmol/L, respectively,and the IC50 of SW1990/GZ cells was（113.08±5.01）μmol/L,（70.26±2.31）μmol/L and （42.12±1.31）μmol/L, respectively.Compared with the untreated cells,the IC50 of gemcitabine in 20,40 ng/μl Latexin treated cells was obviously decreased, and the differences were statistically significant （P〈0.05）.Compared with the SW1990 cells,the expression of Latexin in SW1990/GZ cells was obviously decreased.RI were 37.7, 28.1 and 23.1,respectively.mRNA relative expression of Latexin in SW1990 and SW1990/GZ cells were 0.85±0.08 and 0.31±0.07, and protein relative expression were 0.49±0.09 and 0.13±0.05, and Latexin expression in SW1990/GZ was obviously lower than that in

关 键 词：胰腺肿瘤 Latexin 抗药性 肿瘤 药物疗法 吉西他滨 信号传导 

分 类 号：R735.9[医药卫生—肿瘤]