OBJECTIVETo study the effect of a microRNA-132 antagonist on lithium-pilocarpine-induced status epilepticus (SE) in young Sprague-Dawley (SD) rats.

METHODSForty-five 3-week-old SD rats were randomly and equally divided into epilepticus model group, microRNA-132 antagonist group, and microRNA-132 antagonist negative control group. The young SD rat model of SE was established using lithium-pilocarpine. For the microRNA-132 antagonist group and the negative control group, pretreatment was performed 24 hours before the model establishment. Behavioral observation was performed to assess the latency of SE and success rate of induction of SE. The scale of Lado was used to evaluate the seizure severity. Electroencephalography (EEG) was used to assess the frequency and amplitude of epileptiform discharges. The mortality rate was calculated in each group.

RESULTSThere was no significant difference in the success rate of induction of SE between the three groups (P>0.05). Compared with the microRNA-132 negative control group and the epilepticus model group, the microRNA-132 antagonist group had significantly prolonged SE latency after model establishment (P<0.05), a significantly lower Lado score of seizure (P<0.05), significantly lower frequency and amplitude of epileptiform discharges on EEG (P<0.05), and a slightly reduced mortality rate.

CONCLUSIONSThe treatment with the microRNA-132 antagonist shows an inhibitory effect on the development and progression of lithium-pilocarpine-induced SE in young SD rats. The inhibition of microRNA-132 is likely to be a potential target or direction for drug treatment of SE.

Animals ; Electroencephalography ; Male ; MicroRNAs ; antagonists & inhibitors ; Pilocarpine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Status Epilepticus ; chemically induced ; drug therapy

OBJECTIVETo investigate the effect of microRNA-205 reduction by antagomirs on adhesion ability of normal human corneal epithelial keratinocytes (NHCEKs).

METHODSAntagomir-205, complementary and inhibitory to microRNA-205, was used to suppress endogenous microRNA-205 in NHCEKs. The adhesion ability of treated NHCEKs was then assessed by cell adhesion assay. Immunoblot and immunohistochemistry were conducted to determine the level of two focal adhesion-related proteins, focal adhesion kinase (FAK) and paxillin (Pax). Phalloidin staining was performed to measure the level of filamentous actin in antagomir-treated NHCEKs.

RESULTSAntagomir-205 markedly reduced the level of microRNA-205 in NHCEKs and significantly enhanced adhesion ability of NHCEKs (P<0.01). Further protein analysis validated that inhibition of microRNA-205 increased the number of phosphorylated FAK and phosphorylated Pax, and decreased filamentous actin.

CONCLUSIONOur findings suggest that microRNA-205 has down-regulating effect on cell motility in NHCEKs.

Base Sequence ; Cell Adhesion ; genetics ; Cells, Cultured ; Epithelium, Corneal ; cytology ; Humans ; Keratinocytes ; cytology ; MicroRNAs ; antagonists & inhibitors ; Oligonucleotide Probes

OBJECTIVETo investigate miRNA-221 expression in human colorectal carcinoma (CRC) cells and the effects of miR-221-specific inhibitor on the proliferation and apoptosis of CRC cells.

METHODSFour human CRC cell lines (HT-29, Lovo, SW-480, and CaCO2) were examined for miRNA-221 expression using real-time Q-PCR. The specific 2,-methoxy-modified RNA oligonucleotides of miR-221 (anti-miR-221) were synthesized and transfected into Caco2 cells via liposome, and the changes in the expression of miR-221 in the cells were detected by real-time Q-PCR. The proliferation and apoptosis of the transfected CRC cells were detected using MTT assay and flow cytometry.

RESULTSThe 4 human CRC cells showed significantly upregulated expression of miR-221 compare with HUVECs (P<0.01). The miR-221-specific inhibitor, anti-miR-221, significantly inhibited the expression of miR-221 in Caco2 cells and suppressed the cell proliferation, causing also obvious cell apoptosis (P<0.01).

CONCLUSIONThe miR-221-specific inhibitor shows potent inhibitory effect on the growth of CRC cells, suggesting its value as a potential anti-tumor candidate for treatment of CRC.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colorectal Neoplasms ; pathology ; Humans ; MicroRNAs ; antagonists & inhibitors ; metabolism

OBJECTIVE: To study the effect of microRNA-205 (miRNA-205) on proliferation of laryngeal carcinoma cell line Hep-2. METHOD: The expressions of miRNA-205 in 27 cases laryngeal carcinoma tissues and adjacent normal tissues were detected by Real-time quantitative PCR, the expression of PTEN protein was detected by Western blot. The expressions of PTEN were detected by Western blot after miRNA-205 inhibitor or miRNA-205 mimics was transfected into Hep-2 cells and Hep-2 cells proliferation was measured by CCK-8 kit. RESULT: The expression level of miRNA-205 was significantly higher in laryngeal carcinoma tissues than in adjacent normal tissues (P < 0.01), and the expression of PTEN protein was lower in laryngeal carcinoma tissues than in adjacent normal tissues (P < 0.01). The proliferation rate of Hep-2 cells was decreased significantly and the expression of PTEN protein in Hep-2 cells was increased significantly after miRNA-205 inhibitor was transfected into (P < 0.01), and the proliferation rate of Hep-2 cells was increased significantly and the expression of PTEN protein in Hep-2 cells was decreased significantly after miRNA-205 mimics was transfected into (P < 0.01). CONCLUSION miRNA-205 might promote the proliferation of Hep-2 cells by regulating the expression of PTEN.
Cell Line, Tumor ; Cell Proliferation ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; MicroRNAs ; antagonists & inhibitors ; metabolism ; PTEN Phosphohydrolase ; metabolism ; Transfection

OBJECTIVETo investigate the effects of microRNA (miRNA) on proliferation of cultured human squamous cell carcinoma of tongue Tca8113 cells.

METHODSThe mimics or inhibitors of miRNA-31 or miRNA-139 were transfected into Tca8113 cells using liposome. Tca8113 cell proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay.

RESULTSThe absorbance (A) values of control group at 24 h, 48 h and 72 h were 0.125 +/- 0.002, 0.169 +/- 0.002 and 0.216 +/- 0.004, respectively. The mimics of miRNA-31 increased Tca8113 cell proliferation, with A values increasing to 0.136 +/- 0.001 (P < 0.001), 0.186 +/- 0.004 (P < 0.001) and 0.249 +/- 0.012 (P < 0.01), respectively. The inhibitors of miRNA-139 also increased A values to 0.148 +/- 0.002 (P < 0.001), 0.214 +/- 0.002 (P < 0.001) and 0.250 +/- 0.009 (P < 0.01), respectively. Contrast with these results, the inhibitors of miRNA-31 decreased Tca8113 cell proliferation, with A values decreasing to 0.145 +/- 0.001 and 0.155 +/- 0.011 (both of P < 0.001) at 48 h and 72 h, respectively. The mimics of miRNA-139 also decreased A to 0.135 +/- 0.001 and 0.170 +/- 0.009 (both of P < 0.001).

CONCLUSIONSmiRNA-31 and miRNA-139 play an important role in the carcinogenesis of human tongue carcinomas. It may become a new method for the treatment of tongue carcinomas by adjustment the activities of miRNA.

Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; MicroRNAs ; antagonists & inhibitors ; metabolism ; Tongue Neoplasms ; genetics ; metabolism ; pathology ; Transfection

Translational medicine is a novel concept about combination of basic research and clinical application. The aim of translational medicine is to realize the translation of basic research into clinical practice. microRNAs (miRNAs) are non-coding single-stranded RNAs with 21-25 nucleotides in length as newly discovered factors in regulating gene expression. Recently, the key regulatory role of miRNA in the cardiovascular system has been elucidated and amount of remarkable results has been achieved, particularly in the regulation of cardiac arrhythmias. A series of studies demonstrate that miRNAs are involved in the regulation of expression of a variety of proteins associated with cardiac electrical activity, and are the potential targets of occurrence of cardiac arrhythmias and anti-arrhythmic drugs. miRNAs as a therapeutic target regulate the stability of mRNAs of target genes or play an inhibitory role in the translation process. Stability of the corresponding miRNA expression levels in the target organ may be a new approach for the disease therapy. Regarding the dysfunction of miRNA, we employed miRNA re-expression strategy and anti-miRNA strategy to correct target protein function and provide a new entry for the therapy of arrhythmia. With the technology of miRNA mimics and antagomirs, miRNAs are expected to treat various cardiovascular diseases and will provide a fresh impetus to achieve transform medicine.
Animals ; Anti-Arrhythmia Agents ; therapeutic use ; Arrhythmias, Cardiac ; enzymology ; therapy ; Gene Expression Regulation ; Humans ; Ion Channels ; metabolism ; MicroRNAs ; antagonists & inhibitors ; genetics ; metabolism ; physiology ; RNA Interference ; Translational Medical Research

OBJECTIVETo investigate the effect of anti-miR-145 on human airway smooth muscle cell (HASMC) proliferation and osteopontin systhesis in vitro and explore the mechanisms.

METHODSHASMCs were treated with 10-100 nmol/L anti-miR-145, and the cell proliferation and apoptosis were investigated using a CCK-8 assay and flow cytometry, respectively. The changes in osteopontin synthesis after the treatment was quantified with Western blotting.

RESULTSTreatment with 10 and 50 nmol/L anti-miR-145 significantly promoted the proliferation and osteopontin synthesis in HASMCs (P<0.05 or <0.01), and 50 nmol/L anti-miR-145 obviously inhibited the cell apoptosis (P<0.01).

CONCLUSIONAnti-miR-145 promotes HASMC proliferation and osteopontin synthesis and inhibits HASMC apoptosis in vitro, indicating the important role of anti-miR-145 in the pathogenesis of airway remodeling.

Airway Remodeling ; Apoptosis ; Cell Proliferation ; Cells, Cultured ; Humans ; MicroRNAs ; antagonists & inhibitors ; Myocytes, Smooth Muscle ; drug effects ; Osteopontin ; biosynthesis ; Respiratory System ; cytology

The Wnt signaling pathway plays crucial roles during embryonic development, whose aberration is implicated in a variety of human cancers. Axin, a key component of canonical Wnt pathway, plays dual roles in modulating Wnt signaling: on one hand, Axin scaffolds the "β-catenin destruction complex" to promote β-catenin degradation and therefore inhibits the Wnt signal transduction; on the other hand, Axin interacts with LRP5/6 and facilitates the recruitment of GSK3 to the plasma membrane to promote LRP5/6 phosphorylation and Wnt signaling. The differential assemblies of Axin with these two distinct complexes have to be tightly controlled for appropriate transduction of the "on" or "off" Wnt signal. So far, there are multiple mechanisms revealed in the regulation of Axin activity, such as post-transcriptional modulation, homo/hetero-polymerization and auto-inhibition. These mechanisms may work cooperatively to modulate the function of Axin, thereby playing an important role in controlling the canonical Wnt signaling. In this review, we will focus on the recent progresses regarding the regulation of Axin function in canonical Wnt signaling.
Animals ; Axin Protein ; antagonists & inhibitors ; chemistry ; metabolism ; Epigenesis, Genetic ; Humans ; MicroRNAs ; genetics ; metabolism ; Neoplasms ; genetics ; Protein Processing, Post-Translational ; Wnt Signaling Pathway ; genetics

Microglia are important cells involved in the regulation of neuropathic pain (NPP) and morphine tolerance. Information on their plasticity and polarity has been elucidated after determining their physiological structure, but there is still much to learn about the role of this type of cell in NPP and morphine tolerance. Microglia mediate multiple functions in health and disease by controlling damage in the central nervous system (CNS) and endogenous immune responses to disease. Microglial activation can result in altered opioid system activity, and NPP is characterized by resistance to morphine. Here we investigate the regulatory mechanisms of microglia and review the potential of microglial inhibitors for modulating NPP and morphine tolerance. Targeted inhibition of glial activation is a clinically promising approach to the treatment of NPP and the prevention of morphine tolerance. Finally, we suggest directions for future research on microglial inhibitors.
Humans ; Calcitonin Gene-Related Peptide/antagonists & inhibitors* ; Drug Tolerance ; Hypoglycemic Agents/pharmacology* ; Microglia/physiology* ; MicroRNAs/physiology* ; Minocycline/pharmacology* ; Morphine/pharmacology* ; Neuralgia/etiology* ; Plant Extracts/pharmacology* ; Signal Transduction/physiology*

Animals ; Antineoplastic Agents ; therapeutic use ; Gene Silencing ; Genetic Therapy ; methods ; Humans ; MicroRNAs ; antagonists & inhibitors ; therapeutic use ; Neoplasms ; genetics ; therapy ; RNA Interference ; physiology ; RNA, Small Interfering ; antagonists & inhibitors ; therapeutic use ; RNA-Induced Silencing Complex ; metabolism