槐耳多糖通过抑制AKT/GSK3β/Snail信号通路抑制胰腺癌细胞增殖和上皮间质转化

Huaier polysaccharide inhibits proliferation and epithelial-mesenchymal transition of pancreatic cancer cells by inhibiting AKT/GSK3β/Snail signaling pathway

目的:探讨槐耳多糖(HP)调节丝苏氨酸蛋白激酶B(AKT)/糖原合酶激酶-3β(GSK3β)/锌指转录因子(Snail)信号通路对胰腺癌(PC)细胞增殖、凋亡和上皮间质转化(EMT)的影响。方法:将人胰腺癌SW1990细胞分为:Control组(正常培养)、HP低浓度组(1μg/mL)、HP中浓度组(5μg/mL)、HP高浓度组(10μg/mL)、激活剂组(10μg/mL HP+10μmol/L AKT/GSK3β/Snail通路激活剂SC79);四甲基偶氮唑盐(MTT)法检测细胞增殖;流式细胞仪检测细胞凋亡;黏附实验检测细胞黏附能力;划痕实验检测细胞迁移;Transwell小室检测细胞侵袭;蛋白免疫印迹法(Western blot)方法检测增殖细胞核抗原(PCNA)、Bcl-2相关X蛋白(Bax)、细胞抗凋亡因子B淋巴细胞瘤-2(Bcl-2)、基质金属蛋白酶-2(MMP-2)、波形蛋白(Vimentin)、N-钙黏蛋白(N-cadherin)、E-钙黏蛋白(E-cadherin)及AKT/GSK3β/Snail通路蛋白的表达。结果:与control组比较,HP低、中、高浓度组SW1990细胞OD490值、黏附细胞数、划痕愈合率、侵袭数、PCNA、Bcl-2、MMP-2、Vimentin、N-cadherin、p-AKT/AKT、p-GSK3β/GSK3β、Snail蛋白表达水平降低,细胞凋亡率及Bax、E-cadherin蛋白表达水平升高(P<0.05);与HP高浓度组比较,激活剂组SW1990细胞OD490值、黏附细胞数、划痕愈合率、侵袭数、PCNA、Bcl-2、MMP-2、Vimentin、N-cadherin、p-AKT/AKT、p-GSK3β/GSK3β、Snail蛋白表达水平升高,细胞凋亡率及Bax、E-cadherin蛋白表达水平降低(P<0.05)。结论:HP可能通过抑制AKT/GSK3β/Snail信号通路,抑制SW1990细胞增殖、迁移、侵袭和EMT,促进凋亡。

Objective:To investigate the effects of Huaier polysaccharide(HP)on the proliferation,apoptosis and epithelial-mesenchymal transition(EMT)of pancreatic cancer(PC)cells by regulating the serine-threonine kinase B(AKT)/glycogen synthasc kinase-3β(GSK3β)/zinc finger transcription factor(Snail)signaling pathway.Methods:Human pancreatic cancer SW1990 cells were grouped into control group(normal culture),low concentration HP group(1μg/mL),medium concentration HP group(5μg/mL),high concentration HP group(10μg/mL),activator group(10μg/mL HP+10μmol/L AKT/GSK3β/Snail pathway activator SC79).MTT assay was applied to detect cell proliferation.The apoptosis was detected by flow cytometry.Cell adhesion test was applied to detect cell adhesion.Cell migration was detected by scratch test.Transwell chamber was applied to detect cell invasion,and Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),Bcl-2 associated X protein(Bax),anti apoptotic factor B cell lymphomato-2(Bcl-2),matrix metalloproteinase-2(MMP-2),Vimentin,N-cadherin,E-cadherin and AKT/GSK3β/Snail pathway proteins.Results:Compared with the control group,the SW1990 cell OD490 value,number of adherent cells,scratch healing rate,invasion number,the protein expression levels of PCNA,Bcl-2,MMP-2,Vimentin,N-cadherin,p-AKT/AKT,p-GSK3β/GSK3β,and Snail in the low,medium and high concentration HP groups were lower,the apoptosis rate and the protein expression levels of Bax and E-cadherin were higher(P<0.05).Compared with high concentration HP group,the SW1990 cell OD490 value,number of adherent cells,scratch healing rate,invasion number,the protein expression levels of PCNA,Bcl-2,MMP-2,Vimentin,N-cadherin,p-AKT/AKT,p-GSK3β/GSK3β,and Snail in activator group were higher,the apoptosis rate and the protein expression levels of Bax and E-cadherin were lower(P<0.05).Conclusion:HP may inhibit the proliferation,migration,invasion and EMT of SW1990 cells and promote apoptosis by inhibiting AKT/GSK3β/Snail signaling pathway.