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作 者:庄燕妍[1] 庄晓虹 陈文博[1] 黄凤婷[1] 唐健[1] 程文捷[1] 张世能[1]
机构地区:[1]中山大学孙逸仙纪念医院消化内科,广东广州510120 [2]海南省农垦总医院肿瘤血液科,海南海口571103
出 处:《中山大学学报(医学科学版)》2013年第2期193-199,共7页Journal of Sun Yat-Sen University:Medical Sciences
基 金:广东省自然科学基金(8151008901000139);广东省医学科学基金(B2009066)
摘 要:【目的】探讨3-methyladenine(3-MA)对人胰腺癌SW1990细胞增殖、迁移及失巢凋亡的影响作用及其机制。【方法】以胰腺癌细胞SW1990及永生化胰腺导管上皮细胞H6C7为研究对象,设3-MA处理组及PBS对照组,采用软琼脂克隆培养法及Poly-HEMA悬浮培养法筛选抵抗失巢凋亡的细胞。Annexin V-FITC/PI双染和TUNEL试剂盒法检测细胞凋亡;免疫荧光法及透射电镜检测细胞自噬;CCK-8法检测细胞增殖;Transwell小室检测细胞迁移能力。【结果】永生化胰腺导管上皮细胞H6C7处理组及对照组均无集落形成,胰腺癌细胞SW1990处理组形成(2.3±2.63)个集落,对照组形成(9±3)个,与H6C7相比,SW1990具有较强的抵抗失巢凋亡的能力(P<0.05)。3-MA可抑制胰腺癌SW1990细胞自噬(P<0.05)并抑制其增殖及迁移能力(P<0.01)且诱导失巢凋亡(P<0.01)。【结论】3-MA可诱导胰腺癌细胞SW1990失巢凋亡,其机制可能与细胞自噬受抑有关。[ Objective ] To investigate the effect and mechanism of 3-methyladenine (3-MA) on proliferation, migration, and autophagy of human pancreatic cancer cell line SW1990. [Methods] The experimental objects were pancreatic cancer cell line SW1990 and human immortalized, nontumorigenic pancreatic ductal epithelial cell line H6C7. We divided them into the control group and 3-MA group. Acquisition of anoikis resistance cells was obtained by soft agar clone culture and poly-HEMA suspension culture. Cell apoptosis was detected by Annexin V-FITC/PI and TUNEL. Autophagy assay was measured by immunofluorescence and electronic microscope. Cell proliferation ability was detected by CCK-8. Transwell assay was performed to examine the migration ability. [Results] Compared with H6C7 cells (no clones formation), pancreatic cancer SW1990 cells (2.3 ±2.63) in treatment group, (9 ± 3) in control group showed anoikis resistance (P 〈 0.05). 3-MA could inhibit autophagy of SW1990 ceils under electronic microscope (P 〈 0.05), which could also inhibit the proliferation and migration ability of SW1990 cells (P 〈 0.01) and impair anoikis resistance (P 〈 0.01 ). [Conclusion] 3-MA may induce anoikis of pancreatic cancer cell SW1990 through autophagy inhibition.
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