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<p><b>OBJECTIVE</b>To investigate the possible biological function and mechanism of miR-143 in the metastasis of human nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>Using bioinformatics to predict the target gene of miR-143, the 3'UTR and mutant 3'UTR of GLI3 gene was cloned into psiCHECK-2 vector. Dual-luciferase reporter gene assay was employed to examine the repression of the GLI3 gene. miR-143 and GLI3 expression levels in 5-8F cells transfected with miR-143 mimics, inhibitor, or siGLI3 were examined, and the changes in the cell migration ability was assessed by Transwell invasion assay.</p><p><b>RESULTS</b>Bioinformatics prediction indicated the Hh pathway transcription gene GLI3 as a target gene of miR-143, and dual-luciferase reporter assay showed that miR-143 directly combined with the 3'UTR of GLI3. qRT-PCR and Western blotting demonstrated that the expression of miR-143 in 5-8F cells was negatively correlated to GLI3 and suppressed the migration of 5-8F cells.</p><p><b>CONCLUSION</b>MiR-143 can inhibit the invasion of NPC cells by negative regulation of GLI3 gene, which sheds light on the role of miR-143 and Hh pathway in NPC.</p>
Subject(s)
Humans , Carcinoma , Cell Line, Tumor , Cell Movement , Genetics , Cell Proliferation , Gene Expression Regulation, Neoplastic , Genes, Reporter , Kruppel-Like Transcription Factors , Genetics , MicroRNAs , Genetics , Nasopharyngeal Neoplasms , Genetics , Pathology , Nerve Tissue Proteins , Genetics , Zinc Finger Protein Gli3ABSTRACT
<p><b>OBJECTIVE</b>To detect miR-143 expression in human nasopharyngeal carcinoma (NPC) cell lines and explore the role of miR-143 in regulating the adhesion ability of NPC cells.</p><p><b>METHODS</b>Fluorescence quantitative RT-PCR was used to detect miR-143 expression levels in 5 NPC cell lines (CEN1, CNE2, HONE1, 5-8F, and 6-10B) and an immortalized human nasopharyngeal epithelial cell line (NP69). The retrovirus plasmid pMSCV-puro-miR-143 was constructed and the packaged retroviruses pMSCV-puro and pMSCV-puro-miR-143 were infected in 5-8F cells to establish a cell line with stable miR-143 overexpression, whose adhesion ability was tested with adhesion assay.</p><p><b>RESULTS</b>The expression of miR-143 was down- regulated in the NPC cell lines, and the highly metastatic NPC cell line 5-8F had a expression of only 3.0% of the control level, as compared with the level of 63.59% in the tumorigenic but nonmetastatic NPC cell line 6-10B. The transfected 5-8F cells showed a 1080-fold increase of miR-143 expression (P<0.05) with a significantly lowered adhesion ability.</p><p><b>CONCLUSION</b>miR-143 plays a role in regulating the invasiveness and metastasis of NPC, and overexpression of miR-143 causes a significant reduction of the adhesion ability of the highly metastatic NPC cell line 5-8F.</p>
Subject(s)
Humans , Carcinoma , Cell Adhesion , Cell Line, Tumor , Cell Movement , MicroRNAs , Metabolism , Nasopharyngeal Neoplasms , Metabolism , PathologyABSTRACT
Objective To observe the effect of Shutongan Decoction (STA) on abirritation and generative function of morphine dependence rats and explore its mechanism. Method The model of morphine dependent rat was established. The change of ?-endorphin in hypothalamus and plasma,substance P in ganglion nodosum and nucleus tractus solitarii and incretion hormones in serum of morphine dependence rats were detected by radio-immuno-assay. Result The level of ?-endorphin in hypothalamus of addicted rats was reduced obviously than that in normal rats (P
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Objective To investigate changes in the expression of 5-HT2A receptor mRNA and platelet membrane adhesion molecules in migraineurs during the periods of attack and intermittence therapy by Xinzhengtian Pill (XZTP). To study the interrelation between the expression of 5-HT2A receptor mRNA and platelet membrane adhesion molecules. Methods 68 patients were randomly divided into XZTP (therapy) group and Flunarizine (control) group. The RT-PCR method and the image pattern analysis semiquantitation method were used to determine the 5-HT2A receptor mRNA expression levels in the migraineur groups and the normal control group. Flow cytometry was used to detect the expression of whole blood platelet membrane adhesion molecules in those groups. The interrelation between the expression of 5-HT2A receptor mRNA and adhesion molecules was analysed. Results The total effect have show obvious difference between therapy group and control group. 5-HT2A receptor mRNA in the attack group was higher than that of the intermittence group and the normal group, and XZTP can reduce this expression. The expression of whole blood platelet membrane adhesion molecules had a higher increase during the periods of attack and intermittence, compared with the normal control group and XZTP. The 5-HT2A receptor mRNA expression level in the migraineur groups had a high positive correlation with CD41 and moderate positive correlation with CD62P. Conclusion 5-HT2A receptor mRNA and platelet membrane adhesion molecule expression level during the period of attack was significantly augmented and platelet activation was increased. XZTP maybe restrained the expression of above indexes and resulted in platelet activation reduced.
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[Objective] To investigate the effects of Yiqi Huoxue Tablet (YHT, mainly composed of Radix Angelicae Sinensis, Herba Leonuri, Rhizoma Chuanxiong , Radix Astragali ) on expression of platelet membrane adhesion molecules (PMAM) in whole blood of rats with cerebral infarction. [Methods] Flow cytometry was used to detect the expression of PMAM (CD
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To observe the analgesic effect of Jiedu Tuoyin Decoction (JTD) and its influence on reproduction of morphine dependence rats. JTD is composed of Rhizoma Coptidis, Radix Rehmanniae, Radix Aconiti, Radix Astragali, Radix Codonopsis, Ramulus Uncariae cum Uncis, etc..Sixty rats were randomly allocated to six groups: normal saline group (Group A), morphine and normal saline group (Group B), morphine and clonidine group(Group C),morphine and low dosage of JTD group (Group D), morphine and moderate dosage of JTD group (Group E) and morphine and high dosage of JTD group (Group F).Content of ? endophin(? EP) in hypothalamus and plasma, substance P (SP) content in ganglion nodosum(GN) and nucleus tractus solitarii(NTS) and serum levels of sexual hormones were examined.(1) ? EP content in hypothalamus of morphine dependence rats was lower and SP content in ganglion nodosum and nucleus tractus solitarii was higher than that of normal rats (P
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[Objective] To observe the apoptosis of cortical cells and expression of related gene bcl-2 in hippocampal gyrus and frontal lobe of rats with incomplete cerebral ischemia (ICI) and to investigate the effects of Yiqi Huoxue Tablet (YHT, mainly composed of Radix Astragali , Rhizoma Chuanxiong, Radix Angelicae Sinensis, Herba Leonuri, Rhizoma Acori Tatarinowii, Scorpio, etc) . [ Methods ] Rats were randomized into six groups: model group, mimic operation group, nimodipine group and low-dose, moderate-dose and high-dose YHT groups. Flow cytometry was used to detect the apoptotic rate and bcl-2 protein content in cortical cells. Histological changes were also observed under light microscope. [Results] Early apoptotic changes were found in the model group and the apoptotic rate and bcl-2 protein content were higher than those in the mimic operation group; after treatment of YHT, the apoptotic rate in frontal lobe was decreased as compared with that in the model group and that in hippocampal gyrus remained higher than that in the mimic operation group and bcl-2 protein content was reduced to the level of the mimic operation group. [ Conclusion ] ICI is related to the apoptosis of nerve cells and YHT has been proved an effectual drug for the apoptosis.
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[Objective] To investigate effects Toutong Xin No.l (TX1) on nitric oxide (NO) and nitric oxide synthetase (NOS) levels in platelet and plasma of migraineurs during episode and remission and to explore its mechanism. [Methods] NO and NOS levels in platelet and plasma of migraineurs during episode and remission were measured and were compared with those of healthy volunteers (Group A). [Results] NO and NOS levels in the plasma of migraineurs during episode (Group B) were higher than those during remission (Group C) and Group A (P
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Objective:To investigate the effect of Yishen Bunao Tablet on Alzheimer's Disease(AD).Methods:91 cases of AD were randomly divided into a treatment group(n=46)treated with Yishen Bunao Tablet and a control group(n=45)treated with huperzine A. Their clinical therapeutic effects,changes of superoxide dismutase(SOD)activity,malondialdehyde(MDA)and nitric oxide(NO) contents before and after treatment were determined.Results:There was no significant difference between the two groups in the total therapeutic effect;After treatment,the scores for language comprehend and order obey in the treatment group increased significantly(P0.05).SOD activity increased significantly,and contents of MDA and NO decreased significantly in the control group(P