ABSTRACT
Background:Crohn's disease(CD)was a kind of inflammatory bowel disease,whichwas chronic and recurrent attacked,seriously affect the quality of life of patients.Its pathogenesis is not clear.Aims:To study the effect of hsa_circRNA_103124 which was highly expressed in CD on its downstream gene expression,and the expression level of hsa_circRNA_103124 and its downstream genes in Crohn's disease.The mechanism of hsa_circRNA_103124 in the occurrence and development of CD was discussed in this study.Methods:Transcriptome sequencing revealed differentially expressed genes inhsa_circRNA_103124 overexpressed THP1 cells which was induced macrophage-like differentiation with PMA.IL4 was used to induce macrophage M2 differentiation in hsa_circRNA_103124 overexpressed THP1 cells.The expression levels of M2 differentiation markers CD206 and CD163 were detected by flow cytometry.The expression levels of hsa_circRNA_103124 and its downstream genes were analyzed by qPCR.The levels of hsa_circRNA_103124 and mRNA of its downstream genes in peripheral blood mononuclear cells of 30 patients with CD and 30 healthy controls were analyzed by qPCR.Results:Hsa_circRNA_103124 overexpressed and PMA-induced THP1 cells showed low expression of FGF18(P<0.01).Hsa_circRNA_103124 inhibited macrophage M2 differentiation and down-regulated the expression of CD206(P<0.05)and CD163(P<0.01).The expression of FGF18(P<0.05)and CCL2(P<0.05)was down-regulated in M2-polarized THP1 cells with hsa_circRNA_103124 overexpressed.The expression of hsa_circRNA_103124(P<0.05)in peripheral blood mononuclear cells of CD patients was up-regulated,and the expression of FGF18(P<0.01)and CCL2(P<0.05)was down-regulated.Conclusions:The high expression of hsa_circRNA_103124 in peripheral blood of patients with CD inhibits the M2 polarization of macrophages by down-regulating the expression levels of FGF18 and CCL2,which may play a role in promoting inflammation in the occurrence and development of CD.
ABSTRACT
Objective To explore the diagnostic value of the concentration of plasma circulation microRNA155 (miRNA155) in ulcerative cilitis (UC) and its correlation with clinical characteristic of UC.Methods From October 2010 to August 2012,a total of 136 patients diagnosed as UC were enrolled,and at same time,170 healthy individuals were set as healthy control.The blood samples of all participants were obtained and plasma was isolated.The adsorption column was used for RNA extraction according to miRNeasy kit instruction.RNA was reverse-transcribed into cDNA with miScript reverse transcription kit.cDNA was a template and miRNA155 real-time quantitative polymerase chain reaction (PCR) was performed with miScript SYBR Green PCR kit.The relative quantity of miRNA155 expression was calculated with 2-△△Ct method.Analysis of variance were performed for comparison between groups.Receiver operating characteristic curve analysis was used for the diagnostic value of miRNA155 concentration in UC.Multiple linear regression analysis was used for the correlation between miRNA155 concentration and clinical characteristics of UC.Results The concentration of plasma circulation miRNA155 of patients with UC ((1357.43±326.15) fmol/L)was higher than that of healthy controls ((1140.70 ± 312.47) fmol/L) and the differences were statistically significant (F=35.56,P＜0.01).The area under receiver operating characteristic curve of the concentration of plasma circulation miRNA155 of patients with UC was 0.847,and the 95 ％CI was 0.806 to 0.888 (P＜0.01).When the concentration of plasma circulation miRNA155 was 1404.51 fmol/L,its specificity in the diagnosis of UC was 94.7％,and sensitivity was 40.4％.There was correlation between the concentration of plasma circulation miRNA155 and the disease activity in patients with UC (F=12.91,P＜0.05).However there was no correlation with the severity and location of the disease (both P＞0.05).Conclusion Plasma circulation miRNA155 highly expressed in patients with UC,and its concentration is correlated with the disease activity.
ABSTRACT
Objective To investigate the microRNAs expression profile in human colorectal cancer with or without liver metastasis and try to screen miRNA associated with liver metastasis in colorectal cancer. Methods Twenty five surgical resected colorectal cancer specimens were collected and frozen in liquid nitrogen. Three without liver metastasis and three with liver metastasis were selected, from which total RNAs were isolated. The expressions of miRNAs in these two types of specimens were detected by illumine microRNA microarray, and the difference of miRNA expression was screened. The biochip results were verified with real-time RT-PCR in all colorectal caner specimens. Results The miRNA expression was significantly different in colorectal cancer with liver metastasis and without liver metastasis. Compared with colorectal cancer without liver metastasis, 28 miRNA expressions was different in colorectal cancer with liver metastasis, 4 up regulated and 24 down regulated. The quantity of miR-139-3p expression in colorectal cancer with liver metastasis was 1.75±0.40, up regulated compared with that incolorectal cancer without liver metastasis(0. 69 ±0.58,P＜0.05). The quantity of miR-19a expression in liver metastasis was 0. 39±0. 20, downregulated compare with no liver metastasis( 1.38 ± 0.98, P＜0. 05). The result of miRNA biochip was consistent with that of RT-PCR. Conclusion The difference of miRNA expression might relate to liver metastasis of colorectal cancer. The specific miRNAs expression profile might provide new target for diagnosis and treatment of colorectal cancer with liver metastasis.