奥沙利铂调节LKB1/AMPK/mTOR通路对胃癌细胞增殖、凋亡及免疫逃逸的影响  

Oxaliplatin modulates LKB1/AMPK/mTOR signaling to induce apoptosis and suppress proliferation and immune evasion in gastric cancer cells

作　　者：胡跃云 师琳 张红欣 段亚男 宋惠娟 HU Yueyun;SHI Lin;ZHANG Hongxin;DUAN Ya'nan;SONG Huijuan(Department of Oncology,Shijiazhuang People's Hospital,Shijiazhuang 050000,Hebei,China;Department of GI Medicine,Shijiazhuang People's Hospital,Shijiazhuang 050000,Hebei,China)

机构地区：[1]石家庄市人民医院肿瘤内科,河北石家庄050000 [2]石家庄市人民医院消化内科,河北石家庄050000

出　　处：《贵州医科大学学报》2025年第2期232-239,共8页Journal of Guizhou Medical University

基　　金：河北省卫健委医学科学研究课题计划项目(20251146)。

摘　　要：目的探讨奥沙利铂(oxaliplatin,OXA)调节肝激酶B1(liver kinase B1,LKB1)/单磷酸腺苷激活的蛋白激酶[adenosine 5′-monophosphate(AMP)-activated protein kinase,AMPK]/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)通路对胃癌(gastric cancer,GC)细胞增殖、凋亡及免疫逃逸的影响。方法体外培养人GC细胞(SGC-7901),分为Control组(正常培养)、L-OXA组(6.2 mg/L OXA)、M-OXA组(12.5 mg/L OXA)、H-OXA组(25 mg/L OXA)及Radicicol组(25 mg/L OXA+5μmol/L LKB1抑制剂Radicicol);培养48 h后,采用CCK8和平板克隆实验检测细胞的增殖能力,划痕实验测定细胞的迁移能力,Transwell实验测定细胞的侵袭能力,流式细胞仪测定细胞的凋亡,Western blot测定细胞中PCNA、Bax、Bcl-2、LKB1/AMPK/mTOR信号通路相关蛋白(p-LKB1/LKB1、p-AMPK/AMPK、p-mTOR/mTOR)表达;将GC细胞SGC-7901与CD8^(+)T细胞共培养,并按不同的处理分为共培养组、OXA组、抑制剂组、对照效应细胞(单独培养CD8^(+)T细胞)组及对照靶细胞(单独培养SGC-7901)组,采用CCK8实验计算活化CD8^(+)T细胞的杀伤率。结果与Control组比较,L-OXA组、M-OXA组、H-OXA组细胞的存活率、克隆数、PCNA、迁移率、侵袭数、Bcl-2、p-mTOR/mTOR表达降低,但细胞凋亡率、Bax、p-LKB1/LKB1、p-AMPK/AMPK表达升高(P<0.05);与H-OXA组比较,Radicicol组细胞的存活率、克隆数、PCNA、迁移率、侵袭数、Bcl-2、p-mTOR/mTOR表达升高,但细胞凋亡率、Bax、p-LKB1/LKB1、p-AMPK/AMPK表达降低(P<0.05);免疫逃逸实验中,OXA组较共培养组的活化CD8^(+)T细胞的杀伤率升高(P<0.05);抑制剂组较OXA组的活化CD8^(+)T细胞的杀伤率降低(P<0.05)。结论OXA可能通过调节LKB1/AMPK/mTOR信号通路,促进GC细胞SGC-7901凋亡的发生,抑制其增殖、迁移、侵袭及免疫逃逸。Objective To investigate whether oxaliplatin(OXA)inhibits gastric cancer(GC)cell proliferation,migration,invasion,and immune evasion by regulating the liver kinase B1(LKB1)/adenosine 5′-monophosphate(AMP)-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)pathway.Methods SGC-7901 human GC cells were treated with OXA at low(6.2 mg/L),medium(12.5 mg/L),or high(25 mg/L)concentrations,or with OXA plus the LKB1 inhibitor radicicol(5μmol/L).Cell proliferation was assessed using cell counting kit-8(CCK-8)and colony formation assays.Migration and invasion were evaluated via wound healing and Transwell assays,respectively.Apoptosis was analyzed by flow cytometry.Protein expression[proliferating cell nuclear antigen(PCNA),B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),phosphorylated LKB1(p-LKB1)/LKB1,phosphorylated AMPK(p-AMPK)/AMPK,phosphorylated mTOR(p-mTOR)/mTOR]was measured by Western blot.For immune evasion assays,SGC-7901 cells were co-cultured with activated cluster of differentiation 8-positive(CD8^(+))T cells under OXA or OXA+Radicicol conditions.Cytotoxic activity was quantified using CCK-8.Results OXA dose-dependently reduced cell survival,colony formation,migration,invasion,Bcl-2,and p-mTOR/mTOR levels,while increasing apoptosis,Bax,p-LKB1/LKB1,and p-AMPK/AMPK(P<0.05 vs.Control).Radicicol reversed these effects(P<0.05 vs.H-OXA).OXA enhanced CD8^(+)T cell-mediated cytotoxicity against GC cells(P<0.05 vs.co-culture),which was attenuated by Radicicol(P<0.05).Conclusion OXA suppresses GC progression by activating the LKB1/AMPK/mTOR pathway,promoting apoptosis,and overcoming immune evasion.

关 键 词：奥沙利铂 LKB1/AMPK/mTOR通路 胃肿瘤 细胞增殖 免疫逃逸 

分 类 号：R735.2[医药卫生—肿瘤]