Exosomal MicroRNA-10a Is Associated with Liver Regeneration in Rats through Downregulation of EphA4 / 中华医学杂志(英文版)

<p><b>Background</b>MicroRNAs (miRNAs) have been reported to play vital roles in liver regeneration. Previous studies mainly focused on the functions of intracellular miRNAs, while the functions of circulating exosomal miRNAs in liver regeneration remain largely unknown. The aim of this study was to identify the key exosomal miRNA that played vital roles in liver regeneration.</p><p><b>Methods</b>The Sprague-Dawley male rats were assigned to 70% partially hepatectomized group (n = 6) and sham surgery group (n = 6). The peripheral blood of both groups was collected 24 h after surgery. The exosomal miRNAs were extracted, and microarray was used to find out the key miRNA implicated in liver regeneration. Adenovirus was used to overexpress the key miRNA in rats, and proliferating cell nuclear antigen (PCNA) staining was applied to study the effect of key miRNA overexpression on liver regeneration. Western blotting was used to validate the predicted target of the key miRNA.</p><p><b>Results</b>Exosomal miR-10a was upregulated more than nine times in hepatectomized rats. The level of miR-10a was increased in the early phase of liver regeneration, reached the top at 72 h postsurgery, and decreased to perioperative level 168 h after surgery. Moreover, enforced expression of miR-10a by adenovirus facilitated the process of liver regeneration as evidenced by immunohistochemical staining of PCNA. Erythropoietin-producing hepatocellular receptor A4 (EphA4) has been predicted to be a target of miR-10a. The protein level of EphA4 was decreased in the early phase of liver regeneration, reached the bottom at 72 h postsurgery, and rose to perioperative level 168 h after surgery, which was negatively correlated with miR-10a, confirming that EphA4 served as a downstream target of miR-10a. Moreover, inhibition of EphA4 by rhynchophylline could promote the proliferation of hepatocytes by regulating the cell cycle.</p><p><b>Conclusion</b>Exosomal miR-10a might accelerate liver regeneration through downregulation of EphA4.</p>

miR-10a inhibits cell proliferation and promotes cell apoptosis by targeting BCL6 in diffuse large B-cell lymphoma

The BCL6 (B-Cell Lymphoma 6) gene is a proto-oncogene that is often expressed in diffuse large B-cell lymphomas (DLBCLs). BCL6 loss of function can kill DLBCL cells, demonstrating that BCL6 is necessary for the survival of DLBCL cells and could be a therapeutic target. In this study, we found that BCL6 protein levels were consistently upregulated in DLBCL tissues, whereas its mRNA levels varied randomly in tissues, suggesting that a post-transcriptional mechanism was involved in BCL6 regulation. We used bioinformatics analysis to search for miRNAs, which potentially target BCL6, and identified specific targeting sites for miR-10a in the 3'-untranslated region (3'-UTR) of BCL6. We further identified an inverse correlation between miR-10a levels and BCL6 protein levels, but not mRNA levels, in DLBCL tumor tissue samples. By overexpressing or knocking down miR-10a in DLBCL cells, we experimentally validated that miR-10a directly recognizes the 3'-UTR of the BCL6 transcript and regulated BCL6 expression. Furthermore, we demonstrated that negatively regulating BCL6 by miR-10a suppressed the proliferation and promoted apoptosis of DLBCL cells.

Research progress of miR-10a in tumors / 白血病·淋巴瘤

miR-10a,as one member of miRNA family,belongs to the fannily of miR-10.It locates between HOXB4 and HOXB5 genes on the short arm of chromosome 17.miR-10a is abnormal expressed in a variety of human tumors and is closely related to the occurrence,development and prognosis of tumor.The relationship of miR-10a with a variety of tumors is reviewed in this paper.

DETECTION AND CLINICAL SIGNIFICANCE OF MICRORNA IN OVARIAN CANCER / 现代医院

Objective To study the expressions of miR-10a, miR-93 and miR-200a in malignant ovarian tumor tissues and their clinical significances.Methods Real-time quantitative PCR was used to detect the expression of 40 cases of normal ovarian tissue, 40 cases of benign ovarian cyst and 40 cases of miR-10a, miR-93 and miR-200a in malignant ovarian tumor tissues.Analysis of variance and t-Test were respectively applied to compare the expression of miR in different tissues and analyze the correlation between miR and clinicopathological characteristics of malignant ovarian tumor .Kaplan-Meier Log-rank test and Cox proportional hazards regression model were adopted in prognosis.Results The expression lev-els of miR-10a in ovarian cancer tissues were significantly higher than that in benign ovarian tumor tissues and normal ovari-an tissues (p0.05).The expression quantity of miR-10a in malignant ovarian tumor tissues with greater omen-tum metastasis, lymph node metastasis and distant organ metastasis was significantly higher than that without metastasis ( p<0.05).The median survival time of patients with higher expression of miR -10a was lower than that of patients with low ex-pression of miR-10a (p=0.01).Multi-factor Cox model analysis showed that the expression quantity of miR -10a was an independent factor affecting survival prognosis in patients(p=0.002).Conclusion Our data suggest that miR-10a is asso-ciated with ovarian cancer metastasis, which is the main factor affecting prognosis in ovarian cancer.It might serve as a bio-marker for judging the prognosis in ovarian cancer.

Effects of miR-10a down-regulated by siRNA on migration and invasion of human pancreatic cancer cell AsPC-1 / 中华胰腺病杂志

Objective To investigate the effects of miR-10a expression on migration and invasion of human pancreatic cancer cells AsPC-1.Methods Small interfering RNA targeting at miR-10a (miR-10a-siRNA) was constructed,then it was transfected into pancreatic cancer AsPC-1 cells,and nonsense siRNA (Nc-siRNA) group and blank control group was established.Real time PCR assay was used to detect the expression of miR-10a in the 3 groups,and wound healing assay and Transwell assay were used to determine the migration and invasion abilities of cancer cells.The amount of matrix metalloproteinase-13 (MMP-13) in supernatant of cancer cell culture of each group was examined by ELISA assay.Results The miR-10a levels in control group,NC-siRNA group and miR-10a-siRNA group were 1.05 ±0.08,1.03 ±0.06,0.02 ±0.01 ; and the number of transmembrane cell were (150 ± 2.6),(145 ± 2.2),(62 ± 1.8),the levels of MMP-13 in the supernatant were (108.5 ± 2.8),(107.8 ± 2.5),(35.8 ± 1.5) pg/ml.The values were significantly lower in miR-10a-siRNA group than those in control group and NC-siRNA group (P ＜ 0.01).The distance of cultured clone in miR-10a treated cancer cells (736± 18 μm) was significantly longer than those in the controls (385 ±5 μm) and NC-siRNA group (395± 13 μm,P＜0.01).Conclusions Down-regulation of miR-10a by siRNA may inhibit migration and invasion of pancreatic cancer AsPC-1 cells,and the downregulated expression of MMP-13 may be one of the important mechanisms.