Journal: Cancer Discovery
Article Title: Intrahepatic Microbial Heterogeneity in Multifocal Hepatocellular Carcinoma and Its Association with Host Genomic and Transcriptomic Alterations
doi: 10.1158/2159-8290.CD-24-1259
Figure Lengend Snippet: Microbial distribution of multifocal HCC. A, Flowchart of sample collection and study design. A total of 58 patients with multifocal HCC were enrolled in this study. Tumor nodule samples ( N = 242, 3–11/patient) and adjacent nontumor liver tissue samples ( N = 58) were collected to perform the 16S rRNA gene sequencing, WES, and bulk RNA-seq, followed by integrative analysis and functional validation. B, Alluvial plot depicting the relative abundance of microbial composition at the phylum level (top 5) within each patient. Multiple tumor nodules (right-sided) and adjacent nontumor tissues (left-sided) of each individual are separated by a white line. C, Representative image of Gram staining, bacterial LPS, and LTA staining in the multifocal HCC liver tissue. The quantification of positive signals is presented as mean ± SD on the right. Scale bars, 50 µm (left) and 20 µm (right). “T1–4” refers to tumor nodules. D, Representative images of transmission electron microscopy (TEM) indicating intracellular bacteria (red arrow) within multifocal HCC liver tissue. Scale bars, 2 µm (top) and 0.5 µm (bottom). ( A, Created with BioRender.com .)
Article Snippet: The FFPE slides were deparaffinized, rehydrated, and were stained with bacterial LPS antibody [mAb WN1 222-5; #HM6011; RRID: AB_2750644; 1:200 dilution; Hycult Biotech], bacterial LTA antibody (mAb 55; #HM2048; RRID: AB_57466; 1:400 dilution; Hycult Biotech), E-cadherin (Cell Signaling Technology, Cat# 3195S, 1:1,000, RRID:AB_2291471), N-cadherin (Cell Signaling Technology, Cat# 13116S, 1:1,000, RRID:AB_2687616), Snail (ServiceBio, GB11260, 1:200), and Ki-67 (Cell Signaling Technology, Cat# 12202S, 1:1,000, RRID:AB_2620142).
Techniques: Sequencing, RNA Sequencing, Functional Assay, Biomarker Discovery, Staining, Transmission Assay, Electron Microscopy, Bacteria
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