DataSheet_1_ZNF24 regulates the progression of KRAS mutant lung adenocarcinoma by promoting SLC7A5 translation.zip

BackgroundClinical treatment of RAS mutant cancers is challenging because of the complexity of the Ras signaling pathway. SLC7A5 is a newly discovered downstream gene of the Ras signaling pathway, but the regulatory mechanism is unclear. We aimed to explore the molecular mechanism and role in KRAS mutant lung adenocarcinoma progression.
MethodsKey gene that regulated SLC7A5 in KRAS mutant lung adenocarcinoma was screened by RNA sequencing and bioinformatics analysis. The effect of this gene on the expression of SLC7A5 was studied by RNAi. The regulatory mechanism between the two genes was investigated by immunofluorescence, CoIP, pulldown and yeast two-hybrid assays. The location of the two genes was determined by inhibiting Ras and the downstream pathways PI3K-AKT and MEK-ERK. By in vivo and in vitro experiments, the effects of the key gene on the biological functions of KRAS mutant lung adenocarcinoma were explored.
ResultsWe found a novel gene, ZNF24, which upregulated SLC7A5 protein expression rather than mRNA expression in KRAS mutant lung adenocarcinoma. Endogenous protein interactions occurred between ZNF24 and SLC7A5. Ras inhibition reduced the expression of ZNF24 and SLC7A5. ZNF24 and SLC7A5 are located downstream of the MEK-ERK and PI3K-AKT pathways. In vivo and in vitro functional experiments confirmed that the ZNF24-SLC7A5 signaling axis promoted the proliferation, invasion and migration of KRAS mutant lung adenocarcinoma.
ConclusionsZNF24 promoted the growth of KRAS mutant lung adenocarcinoma by upregulating SLC7A5 protein expression, which suggested that ZNF24 is a new biomarker of KRAS mutant tumors and could be a new potential therapeutic target for Ras-driven tumors.

研究背景：由于Ras信号通路的复杂性，RAS突变型癌症的临床治疗颇具挑战。SLC7A5是新近发现的Ras信号通路下游基因，但其调控机制尚未阐明。本研究旨在探索其在KRAS突变型肺腺癌进展中的分子机制与作用。
研究方法：本研究通过RNA测序与生物信息学分析，筛选KRAS突变型肺腺癌中调控SLC7A5的关键基因；通过RNA干扰（RNAi）实验探究该基因对SLC7A5表达的影响；采用免疫荧光（immunofluorescence）、免疫共沉淀（CoIP）、下拉实验（pulldown）及酵母双杂交（yeast two-hybrid）实验，解析二者间的调控机制；通过抑制Ras及其下游通路PI3K-AKT与MEK-ERK，明确两个基因的上下游定位；通过体内（in vivo）与体外（in vitro）实验，探索该关键基因对KRAS突变型肺腺癌生物学功能的影响。
研究结果：本研究发现全新基因ZNF24，其在KRAS突变型肺腺癌中可上调SLC7A5的蛋白表达，而非mRNA表达；ZNF24与SLC7A5之间存在内源性蛋白质相互作用；抑制Ras可降低ZNF24与SLC7A5的表达水平；ZNF24与SLC7A5均定位于MEK-ERK及PI3K-AKT通路的下游；体内与体外功能实验证实，ZNF24-SLC7A5信号轴可促进KRAS突变型肺腺癌的增殖、侵袭与迁移。
研究结论：ZNF24通过上调SLC7A5的蛋白表达，促进KRAS突变型肺腺癌的生长。这一结果表明，ZNF24可作为KRAS突变型肿瘤的新型生物标志物，同时有望成为Ras驱动型肿瘤的潜在新型治疗靶点。