Trim14 通过调控 AMPK/mTOR 通路促进胃癌细胞自噬和化疗耐药。
Trim14 promotes autophagy and chemotherapy resistance of gastric cancer cells by regulating AMPK/mTOR pathway.
机构信息
Department of Pathology, Seventh People's Hospital of Shanghai University of TCM, Shanghai, China.
Department of Pathology, Shanghai Jiao Tong University, School of Medicine, Shanghai, China.
出版信息
Drug Dev Res. 2020 Aug;81(5):544-550. doi: 10.1002/ddr.21650. Epub 2020 Feb 25.
OBJECTIVE
To study the relationship between TRIM14 expression and chemotherapy resistance of gastric cancer (GC) cells.
METHODS
The expression of TRIM14 in 5-fluorouracil (5-FU)- and oxaliplation (L-OHP)-resistant GC tissues and cells were determined by qRT-PCR and western blotting. PcDNA3.1-TRIM14 and shRNA-TRIM14 vector were transfected to 5-FU-resistant GC cells (SGC7901/5-FU), and the proliferation and apoptosis of cells were measured. Animal experiments on 5-FU-resistant GC mice were performed to study the effect of TRIM14 expression on tumor size and weight, GC cell migration, and proliferation. pcDNA3.1-MK-3903 plasmid was transfected to SGC7901/5-FU cells with TRIM14 silence. The cell proliferation and apoptosis were determined. The protein expressions of Trim14, LC3, and BECLIN1 were measured by western blotting.
RESULTS
TRIM14 was significantly upregulated in 5-FU- and L-OHP-resistant GC tissues and cells. The overexpression of TRIM14 promoted the proliferation and autophagy of SGC7901/5-FU cells, and inhibited the apoptosis. Moreover, in vivo experiment verified that the silence of TRIM14 reduced the tumor size and weight, and inhibited the migration and proliferation of GC cells in 5-FU-resistant GC mice. The overexpression of MK-3903 reversed the inhibiting role of TRIM14 knockout on the proliferation and autophagy of SGC7901/5-FU cells.
CONCLUSION
TRIM14 promoted chemotherapy resistance of GC cells by regulating AMPK/mTOR pathway, and may be a new biomarker for treating GC.
目的
研究 TRIM14 表达与胃癌（GC）细胞化疗耐药性的关系。
方法
采用 qRT-PCR 和 Western blot 检测 5-氟尿嘧啶（5-FU）和奥沙利铂（L-OHP）耐药 GC 组织和细胞中 TRIM14 的表达。将 PcDNA3.1-TRIM14 和 shRNA-TRIM14 载体转染至 5-FU 耐药 GC 细胞（SGC7901/5-FU），检测细胞增殖和凋亡。在 5-FU 耐药 GC 小鼠中进行动物实验，研究 TRIM14 表达对肿瘤大小和重量、GC 细胞迁移和增殖的影响。将 pcDNA3.1-MK-3903 质粒转染至沉默 TRIM14 的 SGC7901/5-FU 细胞，检测细胞增殖和凋亡。采用 Western blot 检测 Trim14、LC3 和 BECLIN1 的蛋白表达。
结果
TRIM14 在 5-FU 和 L-OHP 耐药 GC 组织和细胞中显著上调。TRIM14 的过表达促进了 SGC7901/5-FU 细胞的增殖和自噬，抑制了细胞凋亡。此外，体内实验验证了沉默 TRIM14 可降低 5-FU 耐药 GC 小鼠的肿瘤大小和重量，并抑制 GC 细胞的迁移和增殖。MK-3903 的过表达逆转了 TRIM14 敲除对 SGC7901/5-FU 细胞增殖和自噬的抑制作用。
结论
TRIM14 通过调节 AMPK/mTOR 通路促进 GC 细胞的化疗耐药性，可能成为治疗 GC 的新生物标志物。
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