Expression of integrin αv and CD44v6 in normal hepatocytes with Immunohistochemistry. a,

<p>In <i>IHC</i>, integrin αv proteins (brown positive stain) were localized in cytoplasm of hepatocytes (×400). <b>b,</b> CD44v6 proteins (brown positive stain) were localized in cytoplasm of hepatocytes (×400).</p

OPN protein localization in the CRC and its liver metastasis visualized by immunohistochemistry. a

<p>In <i>IHC</i>, OPN proteins (brown positive stain) were localized in cytoplasm of CRC cells in primary lesions(x200). <b>b</b> Adjacent normal colorectal tissues were negative(x200). <b>c</b> Positive stain was found both in CRC cells and adjacent normal hepatocytes in liver metastatic tissues(x100). <b>d</b> As controls, OPN proteins stain in normal liver tissues without CRC metastasis was negative(x100).</p

OPN mRNA location in CRC and its liver metastasis with in situ mRNA hybridization. a,

<p>In <i>ISH</i>, the positive stain presents blue-purple under microscope. The OPN mRNA, which indicates a positive signal, was found in cytoplasm of CRC cells in primary lesions (×400). <b>b,</b> The stain in adjacent normal colorectal tissues was negative (×400). <b>c,</b> OPN mRNA was found in cytoplasm of CRC cells liver metastatic tissue. Adjacent normal liver tissues stains negative (×100).</p

Gap junctional intercellular communication (GJIC) with gap-FRAP in SW-480-pcDNA3.1(+) and SW-480-pcDNA3.1(+) OPN cells. a1,

<p>uniform fluorescence intensity before photobleaching in SW-480-pcDNA3.1(+). <b>a2,</b> weakened fluorescence intensity after photobleaching in SW-480-pcDNA3.1(+). <b>a3,</b> fluorescence redistribution after 10minutes in SW-480-pcDNA3.1(+). <b>b1,</b> uniform fluorescence intensity before photobleaching SW-480-pcDNA3.1(+) OPN cells. <b>b2,</b> weakened fluorescence intensity after photobleaching SW-480-pcDNA3.1(+) OPN cells. <b>b3,</b> faint fluorescence redistribution after 10minutes SW-480-pcDNA3.1(+) OPN cells. <b>c,</b> The percentage of fluorescence redistribution after photobleaching (FRAP%) in SW-480-pcDNA3.1(+) and SW-480-pcDNA3.1(+) OPN cells<b>.</b> After 5 minutes of photobleaching, the percentage of fluorescence redistribution after photobleaching (FRAP%) 24.65±4.08% in SW-480-pcDNA3.1(+) -OPN compared 44.74±6.23% in SW-480-pcDNA3.1(+)(<i>t</i> = 17.07, <i>P</i><0.001), and after 10 minutes, the FRAP% was still be inhibited at 25.98±4.48% in SW-480-pcDNA3.1(+) -OPN while it was redistributed to 64.92%±5.39% in the SW-480-pcDNA3.1(+) cells (<i>t</i> = 35.16, <i>P</i><0.001).</p

Additional file 1 of Sintilimab plus bevacizumab and CapeOx (BBCAPX) on first-line treatment in patients with RAS mutant, microsatellite stable, metastatic colorectal cancer: study protocol of a randomized, open-label, multicentric study

Additional file 1: Supplementary file 1. A list of participating centers of BBCAPX trial

Exome Capture Sequencing of Adenoma Reveals Genetic Alterations in Multiple Cellular Pathways at the Early Stage of Colorectal Tumorigenesis

<div><p>Most of colorectal adenocarcinomas are believed to arise from adenomas, which are premalignant lesions. Sequencing the whole exome of the adenoma will help identifying molecular biomarkers that can predict the occurrence of adenocarcinoma more precisely and help understanding the molecular pathways underlying the initial stage of colorectal tumorigenesis. We performed the exome capture sequencing of the normal mucosa, adenoma and adenocarcinoma tissues from the same patient and sequenced the identified mutations in additional 73 adenomas and 288 adenocarcinomas. Somatic single nucleotide variations (SNVs) were identified in both the adenoma and adenocarcinoma by comparing with the normal control from the same patient. We identified 12 nonsynonymous somatic SNVs in the adenoma and 42 nonsynonymous somatic SNVs in the adenocarcinoma. Most of these mutations including OR6X1, SLC15A3, KRTHB4, RBFOX1, LAMA3, CDH20, BIRC6, NMBR, GLCCI1, EFR3A, and FTHL17 were newly reported in colorectal adenomas. Functional annotation of these mutated genes showed that multiple cellular pathways including Wnt, cell adhesion and ubiquitin mediated proteolysis pathways were altered genetically in the adenoma and that the genetic alterations in the same pathways persist in the adenocarcinoma. CDH20 and LAMA3 were mutated in the adenoma while NRXN3 and COL4A6 were mutated in the adenocarcinoma from the same patient, suggesting for the first time that genetic alterations in the cell adhesion pathway occur as early as in the adenoma. Thus, the comparison of genomic mutations between adenoma and adenocarcinoma provides us a new insight into the molecular events governing the early step of colorectal tumorigenesis.</p> </div

Validated somatic mutation in the colorectal adenoma.

<p>Validated somatic mutation in the colorectal adenoma.</p

Summary of sequencing coverage of the normal mucosa, adenoma and adenocarcinoma from the same patient.

<p>Summary of sequencing coverage of the normal mucosa, adenoma and adenocarcinoma from the same patient.</p

Somatic SNVs pattern in the adenoma and the adenocarcinoma.

<p>(A) Somatic mutation spectrum in adenoma and adenocarcinoma, similar with 11 colorectal cancers previously reported <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053310#pone.0053310-Sjoblom1" target="_blank">[10]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053310#pone.0053310-Wood1" target="_blank">[11]</a>. (B) Fractions of guanine mutations at 5′-CpG-3′ dinucleotides in the exome of adenoma and adenocarcinoma. (C) Prevalence of somatic SNVs in the coding region and non-coding region of the exome of the adenoma and the adenocarcinoma.</p

Recurrent mutations in 288 additional cases of colorectal tumors.

<p>Recurrent mutations in 288 additional cases of colorectal tumors.</p