MicroRNA Signatures for circulating CD133-positive cells in hepatocellular carcinoma with HCV infection

<div><p>Aim</p><p>Molecular characterization of the CD133+ stem cells associated with hepatocarinogensis through identifying the expression patterns of specific microRNAs (miRNAs).</p><p>Methods</p><p>We investigated the expression pattern of 13 miRNAs in purified CD133+ cells separated from the peripheral blood of healthy volunteers, chronic hepatitis C (CHC), liver cirrhosis (LC) and hepatocellular carcinoma (HCC) patients a long with bone marrow samples from the healthy volunteers and the LC patients using custom miScript miRNA PCR array.</p><p>Results</p><p>The differential expression of the 13 studied miRNAs in CD133+ cells separated from the HCC patients' peripheral blood compared to the controls revealed that <i>miR-602</i>, <i>miR-181b</i>, <i>miR-101</i>, <i>miR-122</i>, <i>miR-192</i>, <i>miR-125a-5p</i>, and <i>miR-221</i> were significantly up regulated (fold change = 1.8, 1.7, 2, 5.4, 1.6, 2.9 & 1.5 <i>P</i> value = 0.039, 0.0019, 0.0013, 0.0370, 00024, 0.000044 &0.000007 respectively). As for the HCC group compared to the CHC group; <i>miR-602</i>, miR-122, <i>miR-181b</i>, <i>miR-125a-5p</i>, and <i>miR-192</i> were significantly up regulated (fold change = 13, 3.1, 2.8, 1.6 & 1.56, <i>P</i> value = 0.01, 0.001, 0.000004, 0.002 & 0.007 respectively). Upon comparing the HCC group to the LC group; <i>miR-199a-3p</i>, <i>miR-192</i>, <i>miR-122</i>, <i>miR-181b</i>, <i>miR-224</i>, <i>miR-125a-5p</i>, and <i>miR-885-5p</i> were significantly up regulated (fold change = 5, 6.7, 2.3, 3, 2.5, 4.2 & 39.5 <i>P</i> value = 0.001025, 0.000024, 0.000472, 0.000278, 0.000004, 0.000075 & 0.0000001 respectively) whereas <i>miR-22</i> was significantly down regulated (fold change = 0.57 <i>P</i> value = 0.00002). Only, <i>miR-192</i>, <i>miR-122</i>, <i>miR-181b</i> and <i>miR-125a-5p</i> were significant common miRNAs in CD133+ cells of the HCC group compared to the other non-malignant groups.</p><p>Conclusion</p><p>We identified a miRNA panel comprised of four miRNAs (<i>miR-192</i>, <i>miR-122</i>, <i>miR-181b</i> and <i>miR-125a-5p</i>) that may serve as a molecular tool for characterization of the CD133+ cells associated with different stages of hepatocarinogensis. This panel may aid in developing a new target therapy specific for those CD133+ cells.</p></div

MicroRNA Signatures for circulating CD133-positive cells in hepatocellular carcinoma with HCV infection - Fig 8

<p>A) Histogram showing the differential expression of the 13 miRNAs in CD133+ cells of the control group (BM) versus the control group (PB). B) Histogram showing the differential expression of the 13 studied miRNAs in CD133+cells of the LC group (BM) versus the LC group (PB). C) Histogram showing the differential expression of the 13 miRNAs in CD133+ cells of the LC group (BM) versus the control group (BM). "*" miRNA is significant at 0.05 level while "**" miRNA is significant at 0.01 level.</p

Functional annotation of miRNAs and their target genes in CD133+ cells of HCC.

<p>Functional annotation of miRNAs and their target genes in CD133+ cells of HCC.</p

MicroRNA Signatures for circulating CD133-positive cells in hepatocellular carcinoma with HCV infection - Fig 3

<p>A) The multi group plot provided both a bar chart and line graph representation (with optional error bars) for differential expression of the13 miRNAs in CD133+ cells of the three groups (PB) compared to control group (PB) where group 1 represented CHC group, group 2 represented LC group and group 3 represented HCC group. B) The heat map with dendrograms representing co-regulated miRNAs across all groups.</p

MicroRNA Signatures for circulating CD133-positive cells in hepatocellular carcinoma with HCV infection - Fig 5

<p>A) Histogram showing the differential expression of the 13 miRNAs in CD133+ cells the CHC group (PB) versus the control group (PB). B) Histogram showing the differential expression of the 13 miRNAs in CD133+cells the LC group (PB) versus the control group (PB). C) Histogram showing the differential expression of the 13 miRNAs in CD133+ cells of the HCC group (PB) versus the control group (PB). D) Histogram showing the differential expression of the 13 miRNAs in CD133+ of the HCC group (PB) versus the CHC group (PB). E) Histogram showing the differential expression of the 13 miRNAs in CD133+ of the HCC group (PB) versus the LC group (PB). "*" miRNA is significant at 0.05 level while "**" miRNA is significant at 0.01 level.</p

The differential expression of the studied miRNAs in the CD133+ cells of the HCC group versus the normal control group.

<p>The differential expression of the studied miRNAs in the CD133+ cells of the HCC group versus the normal control group.</p

MicroRNA Signatures for circulating CD133-positive cells in hepatocellular carcinoma with HCV infection - Fig 2

<p>A) Volcano plot representing the differential expression of the 13 miRNAs in CD133+ cells of the CHC group (PB) versus the control group (PB). B) Volcano plot representing the differential expression of the 13 studied miRNAs in CD133+ cells of the LC group (PB) versus the control group (PB). C) Volcano plot representing the differential expression of the 13 miRNAs in the CD133+ cells of the LC group (PB) versus the control group (PB).</p

Histogram of the confirmatory set showing the differential expression of the four miRNAs in CD133+ cells of the HCC versus the control group (PB).

<p>Histogram of the confirmatory set showing the differential expression of the four miRNAs in CD133+ cells of the HCC versus the control group (PB).</p

Suggested cartoon representing the relationship of significant overlapping miRNAs in CD133+ stem cells associated with hepatocarinogensis on top of HCV infection.

<p>The green color of miRNAs indicates negative fold regulation while and red color indicates positive fold regulation.</p

Circulating Serum miRNAs as Diagnostic Markers for Colorectal Cancer

<div><p>Aim</p><p>The study was designed to assess the possibility of using circulating miRNAs (serum miRNAs) as diagnostic biomarkers in colorectal cancer (CRC) and to identify their possibility as candidates for targeted therapy.</p><p>Methods</p><p>The study involved two sample sets: 1- a training set which included 90 patients with colorectal related disease (30 with CRC, 18 with inflammatory bowel disease (IBD), 18 with colonic polyps (CP) and 24 with different colonic symptoms but without any colonoscopic abnormality who were enrolled as control group) and 2- a validation set which included 100 CRC patients. Serum miRNAs were extracted from all subjects to assess the expression profiles for the following miRNAs (<i>miR-17</i>, <i>miR-18a</i>, <i>miR-19a</i>, <i>miR-19b</i>, <i>miR-20a</i>, <i>miR-21</i>, <i>miR-146a</i>, <i>miR-223</i>, <i>miR-24</i>, <i>miR-454</i>, <i>miR-183</i>, <i>miR-135a</i>, <i>miR- 135b and miR- 92a</i>) using the custom miScript miRNA PCR-based sybergreen array. The area under the receiver operating characteristic curve (AUC) was used to evaluate the diagnostic performance of the studied miRNAs for colorectal cancer diagnosis.</p><p>Results</p><p>Data analysis of miRNA from the training set showed that; compared to control group, only <b><i>miR-19b</i></b> was significantly up-regulated in patients with IBD group (fold change = 5.24, p = 0.016), whereas in patients with colonic polyps, <b><i>miR-18a</i></b> was significantly up-regulated (fold change = 3.49, p-value = 0.018). On the other hand, <i>miR-17</i>, <i>miR-19a</i>, <i>miR-20a and</i> <b><i>miR-223</i></b> were significantly up-regulated (fold change = 2.35, 3.07, 2.38 and 10.35; respectively and p-value = 0.02, 0.015, 0.017 and 0.016; respectively in CRC patients. However, the validation set showed that only <b><i>miR-223</i></b> was significantly up-regulated in CRC patients (fold change = 4.06, p-value = 0.04).</p><p>Conclusion</p><p>Aberrant miRNA expressions are highly involved in the cascade of colorectal carcinogenesis. We have found that (<i>miR-17</i>, <i>miR-19a</i>, <i>miR-20a and miR-223</i>) could be used as diagnostic biomarkers for CRC. On the other hand, <i>miR-19b</i> and <i>miR-18a</i> could be used as diagnostic biomarkers for CP and IBD respectively.</p></div