miR-663a inhibits hepatocellular carcinoma cell proliferation and invasion by targeting HMGA2

Hepatocellular carcinoma (HCC) is a highly aggressive solid malignancy throughout the world. Dysregulation of miRNAs play essential roles in HCC progression via aberrant regulation of cell proliferation, apoptosis, as well as metastasis. miR-663a is a poorly investigated miRNA. Whether miR-663a regulates HCC development remains unknown. The aim of the study was to explore the role of miR-663a in HCC development. To determine the expression level of miR-663a in HCC, we analyzed the data from GSE21362 and TCGA. The results showed that miR-663a was significantly down-regulated in HCC tissue compared with adjacent non-tumor tissue. Gain of function and loss of function assays revealed that miR-663a distinctly inhibited cell proliferation, migration and invasion. Mechanistic investigations demonstrated that miR-663a modulated cell functions through targeting and suppressing high mobility group A2 (HMGA2). In addition, overexpression of HMGA2 remarkably attenuated the tumor repressive effect of miR-663a. Taken together, miR-663a inhibits HCC cell proliferation and motility by targeting HMGA2.     

microRNA-622 acts as a tumor suppressor in hepatocellular carcinoma

microRNAs (miRNAs) are important regulators of tumor development and progression. In this study, we aimed to explore the expression and role of miR-622 in hepatocellular carcinoma (HCC). We found that miR-622 was significantly downregulated in human HCC specimens compared to adjacent noncancerous liver tissues. miR-622 downregulation was significantly associated with aggressive parameters and poor prognosis in HCC. Enforced expression of miR-622 significantly decreased the proliferation and colony formation and induced apoptosis of HCC cells. In vivo studies demonstrated that miR-622 overexpression retarded the growth of HCC xenograft tumors. Bioinformatic analysis and luciferase reporter assays revealed that miR-622 directly targeted the 3′-untranslated region (UTR) of mitogen-activated protein 4 kinase 4 (MAP4K4) mRNA. Ectopic expression of miR-622 led to a significant reduction of MAP4K4 expression in HCC cells and xenograft tumors. Overexpression of MAP4K4 partially restored cell proliferation and colony formation and reversed the induction of apoptosis in miR-622-overexpressing HCC cells. Inhibition of JNK and NF-κB signaling phenocopied the anticancer effects of miR-622 on HCC cells. Taken together, miR-622 acts as a tumor suppressor in HCC and restoration of miR-622 may provide therapeutic benefits in the treatment of HCC.     

CircKIF4A靶向调控miR-384表达对卵巢癌SKOV3细胞增殖和凋亡的影响

Objective?To explore the effect of circKIF4A on the proliferation and apoptosis of ovarian cancer SKOV3 cells and its regulatory mechanism on miR-384. Methods?The qRT-PCR method was used to detect the expression of circKIF4A and miR-384 in ovarian cancer tissues and adjacent tissues. si-NC, si-circKIF4A, miR-NC, miR-384 mimics, si-circKIF4A and anti-miR-NC, si-circKIF4A and anti-miR-384 were transfected into SKOV3 cells respectively. The qRT-PCR method was used to detect the expression of circKIF4A and miR-384 in SKOV3 cells. MTT experiment and flow cytometry experiment were used to detect cell proliferation and apoptosis respectively. The dual luciferase reporter experiment was used to detect the targeting relationship between circKIF4A and miR-384. Results?Compared with adjacent tissues, the expression level of circKIF4A in ovarian cancer tissues was increased [(1.00±0.06), (4.28±0.32)] (t=62.915, P＜0.05), and the expression level of miR-384 was decreased [(1.00± 0.05), (0.43±0.03)] (t=61.047, P<0.05). After transfection with si-circKIF4A, the OD value of SKOV3 cells was decreased [(0.75±0.05), (0.41±0.03)] (t=17.493, P＜0.05), and the apoptosis rate was increased [(6.36±0.53)%, (23.19± 2.21)%] (t=22.216, P<0.05). After transfection of miR-384 mimics, the OD value of SKOV3 cells was decreased [(0.73±0.05), (0.47±0.04)] (t=12.182, P＜0.05), and the apoptosis rate was increased [(7.53±0.41)%, (19.11)±1.06)%] (t=30.567, P<0.05). The dual luciferase report experiment confirmed that circKIF4A could adsorb miR-384 and can act as a sponge molecule for miR-384. After co-transfection with si-circKIF4A and anti-miR-384, the OD value of SKOV3 cells was increased [(0.40±0.04), (0.65±0.03)] (t=15.000, P＜0.05), and the apoptosis rate was decreased [(25.20± 2.21)%, (10.37±0.86)%] (t=18.761, P＜0.05). Conclusion?Inhibition of circKIF4A expression could negatively regulate the expression of miR-384, thereby inhibiting the proliferation of ovarian cancer cells and inducing apoptosis.     

RNA Biomarker Release with Ultrasound and Phase-Change Nanodroplets

Microbubbles driven by ultrasound are capable of permeabilizing cell membranes and allowing biomarkers or therapeutics to exit from or enter cancer cells, respectively. Unfortunately, the relatively large size of microbubbles prevents extravasation. Lipid-based perfluorobutane microbubbles can be made seven-fold smaller by pressurization, creating 430-nm nanodroplets. The present study compares microbubbles and nanodroplets with respect to their ability to enhance miR-21 and mammaglobin mRNA release from cultured ZR-75-1 cells. Mammaglobin mRNA and miR-21 release increased with escalating concentrations of nanodroplets up to, respectively, 25- and 42-fold with 2% nanodroplets (v/v), compared with pre-ultrasound levels, whereas cell viability decreased to 62.4%. Sonication of ZR-75-1 cells incubated with microbubbles or nanodroplets caused relatively similar levels of cell death and miR-21 release, suggesting that nanodroplets are similar to microbubbles in enhancing cell permeability, but may be more advantageous because of their smaller size, which may allow extravasation through leaky tumor vasculature.     

Down-regulation of microRNA-27b promotes retinal pigment epithelial cell proliferation and migration by targeting Nox2

Aberrant proliferation and migration of retinal pigment epithelium (RPE) cells contributes to the pathology of various ocular diseases. miR-27b has been reported to be crucial in the regulation of cell differentiation, proliferation, apoptosis, and migration. However, the role of miR-27b on RPE proliferation and migration remains largely unknown. Here the effect of miR-27b on ARPE-19 cells under platelet-derived growth factor (PDGF)-BB stimulation was explored. In this study, we found that the expression level of miR-27b was significantly reduced in ARPE-19 cells under PDGF-BB stimulation. Ectopic expression of miR-27b remarkably inhibited PDGF-BB-induced proliferation and migration in ARPE-19 cells. Furthermore, bioinformatic analysis and luciferase reporter assay showed that NADPH oxidase 2 (Nox2) was a direct target for miR-27b, and that knockdown of Nox2 expression mimicked the inhibitory effect of miR-27b on PDGF-BB ?induced proliferation and migration in ARPE-19 cells, whereas, restoration of Nox2 expression showed an opposite effect. In addition, the ROS production and the activation of P13K/AKT/mTOR signaling induced by PDGF-BB were also suppressed by miR-27b overexpression or Nox2 silencing. Thus, these findings indicated that miR-27b exerted its protective role in RPE cells under PDGF-BB stimulation was partially through regulation of Nox2 and its downstream P13K/AKT/mTOR signaling, which might be a potential therapeutic approach for treatment of diseases caused by RPE proliferation, and migration.     

狼疮性肾炎肾血管损害和miR-145的表达

目的评估狼疮性肾炎(LN)患儿肾血管损害(RVLs)并检测肾血管中mi R-145的表达。方法收集41例经肾活检证实为LN的系统性红斑狼疮(SLE)患儿的临床资料,评估肾小球损害和RVLs。根据RVLs评分及病理类型分组。采用原位杂交法检测mi R-145在肾血管中的表达。比较不同病理分型组间RVLs、肾血管mi R-145表达及肾小球损害评分差异;比较不同RVLs组间临床指标、肾小球损害评分及肾血管mi R-145表达差异。结果不同LN病理类型间,RVLs的差异无统计学意义(P0.05),肾血管mi R-145表达及肾小球损害评分差异有统计学意义(P0.01)。不同RVLs组间,临床指标及肾小球损害差异均无统计学意义(P0.05),而肾血管mi R-145表达差异有统计学意义(P0.01)。结论 LN患儿存在RVLs,mi R-145可能参与RVLs的发生机制。     

肝硬化患者血清miR-122的表达及与Child-Pugh分级的关系

目的探讨肝硬化患者血清MicroRNA(miR)-122的表达水平与肝硬化Child-Pugh分级及并发症的关系。方法肝硬化患者87例,采用PCR实时荧光定量法测定患者血清miR-122水平,根据Child-Pugh分级对肝硬化患者进行分级,分析血清miR-122与肝硬化分级的关系。结果并发肝硬化腹水、消化道出血、自发性细菌性腹膜炎及失代偿期患者血清miR-122表达水平均高于未发生患者(P<0.05);不同肝硬化Child-Pugh分级血清中miR-122水平有统计学差异(P<0.05),A级与B级肝硬化患者血清miR-122水平无显著差异(P>0.05),A级和B级患者水平均显著低于C级(P<0.05)。多变量线性相关性显示,血清miR-122与丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、γ-谷氨酰转移酶(GGT)、碱性磷酸酶(ALP)呈正相关(P<0.05);与国际标准化比值(INR)及肌酐(Cr)水平呈负相关(P<0.05)。结论血清miR-122水平上升提示肝硬化已经失代偿,并与腹水、消化道出血、肝肾衰竭有关。因此,血清miR-122可作为评价肝硬化患者肝脏功能及预后的潜在生物学指标。     

MiR-876-5p acts as an inhibitor in hepatocellular carcinoma progression by targeting DNMT3A

Hepatocellular carcinoma (HCC) is one of the biggest challenges that human beings faced with in 21st century. Previous researches have revealed that miRNAs can serve as regulators in various cancers. MiR-876-5p, a member of miRNA family, has been studied in lung cancer for its anti-oncogenic function. However, the exact function of it is not reported in HCC. Our study aims to find out the effects of miR-876-5p expression on HCC progression. Two HCC cells were chosen to do functional assays after miR-876-5p expression was detected in cell lines by qRT-PCR. HepG2 cell was transfected with miR-876-5p mimics, whereas LM3 cell was transfected with miR-876-5p inhibitors. Next, cell activities of these two indicated cells were analyzed by means of MTT assay, colony forming assay, transwell migration assay and western blot analysis. Consequently, we found that miR-876-5p could inhibit both cell proliferation and metastasis. Moreover, we found out a target gene (DNMT3A) of miR-876-5p by performing bioinformatics analysis, dual luciferase reporter assay and biotin-avidin pull-down assay. Finally, rescue assays were carried out in HepG2 cells. We found that DNMT3A could reverse miR-876-5p mimics-induced inhibition. Therefore, we concluded that miR-876-5p suppressed hepatocellular carcinoma progression by targeting DNMT3A.     

mTOR负调控miR-27a并通过靶向降低GP73抑制人肝癌细胞侵袭

目的探索m TOR促进肝癌细胞侵袭能力机理。方法使用q-PCR方法检测miR-27a和GP73表达;将miR-27a的mimic转染GP73高表达的M97H细胞中,并将miR-27a inhibitor转入GP73低表达Hep G2细胞中,q-PCR和Western blot观察GP73的表达;使用荧光素酶报告基因系统验证miR-27a在GP73的3UTR区的结合位点;将miR-27a mimic转染GP73高表达的M97H细胞和GP73低表达的Hep G2细胞,Transwell实验观察细胞的侵袭。结果m TOR下调miR-27a,并上调GP73的表达;miR-27a下调GP73的表达,抑制miR-27a则上调GP73;GP73是miR-27a的靶基因;miR-27a显著抑制M97H细胞的侵袭,但对Hep G2细胞无抑制效果。结论 m TOR负调控的miR-27a靶向GP73抑制肝癌细胞的侵袭。