摘要
目的:研究TGF-α是否具有EGF相似的可诱导前列腺癌DU145细胞发生神经内分泌分化(neuroendocrine differentiation,NED)的作用,并探讨TGF-α诱导的前列腺癌DU145细胞神经内分泌分化对肿瘤化疗耐药性的影响。方法:将接受不同培养液处理的DU145细胞分为3组:2%FBS组、2%FBS+TGF-α5 ng/mL组和2%FBS+TGF-α10 ng/mL组。显微镜下观察TGF-α处理后DU145细胞的形态变化;Real time RT-PCR法检测神经特异性烯醇化酶(neuron-specific enolase,NSE)mRNA的表达水平;Western印迹检测NSE,P-糖蛋白(P-glycoprotein,P-gp),多药耐药相关蛋白1(multiple drug resistance protein,MRP1)和Bcl-2蛋白的表达水平。流式细胞术分析TGF-α(5μg/mL)处理后DU145细胞周期的变化;MTT比色法测定TGF-α(5μg/mL)对DU145细胞顺铂化疗耐药性的影响。结果:同2%FBS组相比,2%FBS+TGF-α处理组的DU145细胞出现多形性,细胞伪足伸出;NED标志物NSEmRNA的表达水平升高,分别为上调了(3.6±0.5)倍(P<0.05)和(10.1±0.1)倍(P<0.01);细胞的NSE,Bcl-2和MRP1蛋白的表达水平也明显增加,但未检测到P-gp蛋白的表达;同时,TGF-α(5μg/mL)处理后DU145细胞的G1期细胞比例降低,S期和G2/M期细胞比例升高;顺铂的化疗耐药性增加。结论:TGF-α也可诱导前列腺癌DU145细胞神经内分泌分化增强,从而进一步增强DU145细胞对顺铂的化疗耐药性。
Objective： To study whether TGF-α possesses similar EGF effect of enforcing neuroendocrine differentiation （NED） in prostate cancer cell line DU145 and determine the influence of NED induced by TGF-α on chemoresistance. Methods： DU145 cells were divided into 3 groups： a group with 2% FBS, a group with 2%FBS＋TGF-a S ng/mL and a group with 2%FBS＋TGF-a 10 ng/mL. Morphological change in DUI4S cells was observed after TGF-α treatment. Expression levels of NSE mRNA were detected with real time RT-PCR. Western blot was used to detect the expression levels of protein NSE, P-gp, MRP1 and Bcl-2. Cell cycles of DU145 cells in the 3 groups were examined with flow cytometry. MTT assay was used to evaluate the influence of TGF-α in chemoresistance. Results： Compared with DU145 cells cultured with 2% FBS, cells treated with 2% FBS＋TGF-α were pleomorphic and pseudopodia extended. The expression level of NSE mRNA upregulated to （3.6±0.5） folds （P〈0.OS） and （10.1±0.1） folds （P〈0.01）. Western blot showed that the expression levels of protein NSE, Bcl-2, and MRP1 increased after treatment with different concentrations of TGF-α; P-gp was not detected. The proportion of DU145 cells in phase G1 decreased; proportions of cells in phase S and phase G2/M were increased after TGF-α treatment （5μtg/mL）. At the same time, chemoresistance of DU145 cells to cisplatin increased. Conclusion： TGF-α can increase NED in DU145 cells and enforce the chemoresistance to cisplatin.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2013年第2期142-147,共6页
Journal of Central South University ：Medical Science
基金
湖南省卫生厅科研基金课题(132009-007)~~
关键词
前列腺癌DU145细胞
转化生长因子
神经内分泌分化
prostate cancer cell line DU145
transforming growth factor
neuroendocrine differentiation
