Introduction:Recent studies have discovered that lung cancer subtypes possess distinct microbiome profiles within their tumor microenvironment. Additionally, the tumor-associated microbiome exhibits altered bacterial pathways, suggesting that certain bacterial families are more capable of facilitating tumor progression than others. Wehypothesizethat there exists a crosstalk between lung adenocarcinoma (LUAD) cells and bacterial cells.Methods and Materials:RNA sequencing (RNA-seq) was performed on LUAD cell lines to explore the paracrine signaling effects of bacterial biomolecules. Based on our RNA-seq data, we investigated glycolysis by measuring glucose uptake and lactate production, invasive potential through invasion assays, and epithelial-to-mesenchymal transition (EMT) markers. Since lipopolysaccharides (LPS), abundant on the cell walls of Gram-negative bacteria, can activate toll-like receptor 4 (TLR4), we inhibited TLR4 with C34 to assess its relationship with the observed phenotypic changes. To identify the bacterial biomolecules responsible for these changes, we treated the media with RNAse enzyme, charcoal or dialyzed away molecules larger than 3 kDa.Results and Discussion:RNA-seq revealed 948 genes upregulated in the presence ofE. colibiomolecules. Among these, we observed increased expression of Hexokinase II (HKII), JUN proto-oncogene, and Snail Family Transcriptional Repressor 1. We verified the elevation of glycolytic enzymes through Western blot and saw elevation of 2-deoxyglucose uptake and lactate production in LUAD cell lines incubated inE. colibiomolecules. In addition toE. colielevating glycolysis in LUAD cell lines,E. coliexposure enhanced invasive potential as demonstrated by Boyden chamber assays. Notably, inhibition of TLR4 did not reduce the impact ofE. colibiomolecules on glycolysis or the invasive potential of LUAD. Modulating theE. coli-supplemented media with RNAse enzyme or dextran-coated charcoal or using a spin column to remove biomolecules smaller than 3 kDa resulted in changes in HKII and Claudin protein expression. These findings suggest a direct relationship betweenE. coliand LUAD, wherein several cancer hallmarks are upregulated. Future studies should further investigate these bacterial biomolecules and their role in the tumor microenvironment to fully understand the impact of microbial shifts on cancer progression.
引言：近期研究发现，肺癌亚型在肿瘤微环境中具有独特的微生物组特征。此外，肿瘤相关微生物组表现出细菌通路的改变，表明某些细菌家族比其他细菌更能促进肿瘤进展。我们推测肺腺癌细胞与细菌细胞之间存在交互作用。 方法与材料：对肺腺癌细胞系进行RNA测序，以探究细菌生物分子的旁分泌信号效应。基于RNA测序数据，我们通过测量葡萄糖摄取和乳酸产量研究糖酵解，通过侵袭实验评估侵袭潜能，并检测上皮-间质转化标志物。由于革兰氏阴性菌细胞壁富含的脂多糖可激活Toll样受体4，我们使用C34抑制该受体以评估其与表型变化的关系。为确定导致这些变化的细菌生物分子，我们分别用RNA酶、活性炭处理培养基，或透析去除大于3 kDa的分子。 结果与讨论：RNA测序显示大肠杆菌生物分子存在时948个基因表达上调，其中己糖激酶II、JUN原癌基因及Snail家族转录抑制因子1表达增加。通过Western blot验证糖酵解酶升高，并观察到经大肠杆菌生物分子孵育的肺腺癌细胞系中2-脱氧葡萄糖摄取和乳酸产量增加。除增强糖酵解外，Boyden小室实验证实大肠杆菌暴露可提升肺腺癌细胞的侵袭潜能。值得注意的是，抑制Toll样受体4并未减弱大肠杆菌生物分子对糖酵解或侵袭潜能的影响。使用RNA酶、葡聚糖包被活性炭处理培养基，或通过离心柱去除小于3 kDa的生物分子，均导致己糖激酶II和紧密连接蛋白表达改变。这些发现揭示了大肠杆菌与肺腺癌之间存在直接关联，其中多个癌症特征标志被上调。未来研究需进一步探究这些细菌生物分子及其在肿瘤微环境中的作用，以全面理解微生物变化对癌症进展的影响。
E. coliBiomolecules Increase Glycolysis and Invasive Potential in Lung Adenocarcinoma
肿瘤(癌症)患者之家