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

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 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

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 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

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