阻断lncRNA-SNHG16通过靶向mir -506-3p- ptbp1介导的葡萄糖代谢使胃癌细胞对5-Fu增敏。

IF 6 3区 医学 Q1 CELL BIOLOGY
Yan Ding, Sujie Gao, Jiabin Zheng, Xuebo Chen
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引用次数: 1

摘要

背景:胃癌是一种常见的人类恶性肿瘤。5-氟尿嘧啶(5-Fu)是一线抗胃癌药物。然而,大量GC患者出现5-Fu耐药。目前,lncrna - snhg16调控的5-Fu耐药在胃癌中的作用和分子机制尚不明确。方法:采用qRT-PCR和Western blot检测lncRNA、miRNA和mRNA的表达。采用RNA下拉法和荧光素酶法检测RNA-RNA相互作用。MTT法和Annexin V法检测5-Fu处理下细胞活力和凋亡率。葡萄糖摄取和ECAR测定GC细胞的糖酵解速率。结果:我们报道了SNHG16和rna结合蛋白PTBP1与5-Fu对胃癌的耐药呈正相关。SNHG16和PTBP1在胃肿瘤和细胞系中显著上调。沉默SNHG16或PTBP1可有效使GC细胞对5-Fu敏感。此外,葡萄糖代谢在5- fu耐药的GC细胞中再次显著升高。在低糖条件下,5- fu耐药细胞比亲本GC细胞表现出更高的脆弱性。生物信息学分析和荧光素酶测定表明,SNHG16通过海绵化miR-506-3p形成ceRNA网络,从而下调miR-506-3p。我们发现PTBP1是GC细胞中miR-506-3p的直接靶点。RNA-seq结果显示,PTBP1正调控多种糖酵解酶的表达,包括GLUT1、HK2和LDHA。生物信息学分析表明,糖酵解酶的3′utr含有多个PTBP1结合位点,并通过RNA pull-down和RNA免疫沉淀实验进一步验证。因此,我们证明了PTBP1通过促进糖酵解酶mRNA的稳定性来上调糖酵解酶的mRNA。最后,体内异种移植实验证实,阻断snhg16介导的miR-506-3p-PTBP1轴可有效限制5-Fu治疗下5-Fu耐药GC细胞源异种移植肿瘤的生长。结论:我们的研究通过调节mir -506-3p- ptbp1 -葡萄糖代谢轴,揭示了snhg16介导的GC细胞5-Fu耐药的分子机制,为抗化疗耐药提供了一条有前景的途径。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Blocking lncRNA-SNHG16 sensitizes gastric cancer cells to 5-Fu through targeting the miR-506-3p-PTBP1-mediated glucose metabolism.

Blocking lncRNA-SNHG16 sensitizes gastric cancer cells to 5-Fu through targeting the miR-506-3p-PTBP1-mediated glucose metabolism.

Blocking lncRNA-SNHG16 sensitizes gastric cancer cells to 5-Fu through targeting the miR-506-3p-PTBP1-mediated glucose metabolism.

Blocking lncRNA-SNHG16 sensitizes gastric cancer cells to 5-Fu through targeting the miR-506-3p-PTBP1-mediated glucose metabolism.

Background: Gastric cancer (GC) is a commonly occurring human malignancy. The 5-fluorouracil (5-Fu) is a first-line anti-gastric cancer agent. However, a large number of GC patients developed 5-Fu resistance. Currently, the roles and molecular mechanisms of the lncRNA-SNHG16-modulated 5-Fu resistance in gastric cancer remain elusive.

Methods: Expressions of lncRNA, miRNA, and mRNA were detected by qRT-PCR and Western blot. RNA-RNA interaction was examined by RNA pull-down and luciferase assay. Cell viability and apoptosis rate under 5-Fu treatments were determined by MTT assay and Annexin V assay. The glycolysis rate of GC cells was evaluated by glucose uptake and ECAR.

Results: Here, we report that SNHG16 as well as PTBP1, which is an RNA-binding protein, are positively associated with 5-Fu resistance to gastric cancer. SNHG16 and PTBP1 were significantly upregulated in gastric tumors and cell lines. Silencing SNHG16 or PTBP1 effectively sensitized GC cells to 5-Fu. Furthermore, glucose metabolism was remarkedly elevated in 5-Fu-resistant GC cells. Under low glucose supply, 5-Fu-resistant cells displayed higher vulnerability than parental GC cells. Bioinformatic analysis and luciferase assay demonstrated that SNHG16 downregulated miR-506-3p by sponging it to form a ceRNA network. We identified PTBP1 as a direct target of miR-506-3p in GC cells. RNA-seq results unveiled that PTBP1 positively regulated expressions of multiple glycolysis enzymes, including GLUT1, HK2, and LDHA. Bioinformatic analysis illustrated the 3'UTRs of glycolysis enzymes contained multiple PTBP1 binding sites, which were further verified by RNA pull-down and RNA immunoprecipitation assays. Consequently, we demonstrated that PTBP1 upregulated the mRNAs of glycolysis enzymes via promoting their mRNA stabilities. Finally, in vivo xenograft experiments validated that blocking the SNHG16-mediated miR-506-3p-PTBP1 axis effectively limited 5-Fu-resistant GC cell originated-xenograft tumor growth under 5-Fu treatments.

Conclusions: Our study demonstrates molecular mechanisms of the SNHG16-mediated 5-Fu resistance of GC cells through modulating the miR-506-3p-PTBP1-glucose metabolism axis, presenting a promising approach for anti-chemoresistance therapy.

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来源期刊
自引率
1.70%
发文量
17
审稿时长
14 weeks
期刊介绍: Cancer & Metabolism welcomes studies on all aspects of the relationship between cancer and metabolism, including: -Molecular biology and genetics of cancer metabolism -Whole-body metabolism, including diabetes and obesity, in relation to cancer -Metabolomics in relation to cancer; -Metabolism-based imaging -Preclinical and clinical studies of metabolism-related cancer therapies.
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