3-methyladenine抑制自噬诱导胰腺癌细胞SW1990失巢凋亡

3-Methyladenine Impairs Anoikis Resistance of Pancreatic Cancer SW1990 Cells Through Inhibiting Autophagy In Vitro

【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.