Effect of Mir-127-3p on Proliferation，Apoptosis，Migration and Invasion of Uveal Melanoma Cells via Targeting MAPK4 / 中山大学学报(医学科学版)

@#【Objective】To investigate the effect of miR- 127-3p on proliferation，apoptosis，migration and invasion of uveal melanoma cells.【Methods】The expression of miR- 127- 3p and MAPK4 mRNA in human uveal melanoma tissues and cells，normal tissues and cells were detected by RT-qPCR. The mimic-NC，miR-127-3p mimic，pc-MAPK4 plasmids were transfected into SP6.5 or OM431 cells，respectively，by Lipofectamine 2000. The relationship between miR-127-3p and MAPK4 counterstaining was detected by dual luciferase assay. Cell proliferation was detected by CCK- 8 method，apoptosis was detected by flow cytometry，cell migration ability was detected by scratch test，cell invasion ability was detected by Transwell method，and relative expression level of AKT/mTOR pathway protein was detected by Western blot.【Results】In uveal melanoma tissues and cell lines，the expression of miR- 127-3p was down-regulated（P < 0.01）while that of MAPK4 expression was significantly up-regulated（P < 0.01）. The binding site of miR-127-3p and MAPK4 3′UTR region，the high expression of miR-127-3p significantly inhibited the luciferase activity of wild-type MAPK4 plasmid（P < 0.01），but the mutant MAPK4 plasmid Luciferase activity has no effect. Compared with the Control group ，the proliferation of SP6.5 cells and OM431 cells in miR- 127-3p mimic group were significantly decreased（P < 0.01），and the apoptotic rate was significantly increased（P < 0.01）. The scratch closure rate was obvious. The decrease（P < 0.01）， the number of invading cells per field was significantly decreased（P < 0.01），and the expression of p-AKT（T308）/AKT and p-mTORr（S473）/mTOR protein were significantly down-regulated（P < 0.01）. Transfection of pc-MAPK4 reversed the above changes.【Conclusion】MiR-127-3p inhibits proliferation，migration and invasion of uveal melanoma cells and induces apoptosis by down-regulating MAPK4，which may be involved in the inhibition of AKT/mTOR pathway activation.

A Pilot Study for Discovering Candidate Genes of Chromosome 18q21 in Methamphetamine Abusers: Case-control Association Study

OBJECTIVE: It was previously suggested that the malic enzyme 2 (ME2) as the candidate gene for psychosis in fine mapping of chromosome 18q21. Chromosome 18q21 is also one of the possible regions that can contribute to addiction. METHODS: We performed a pilot study for discovering candidate gene of chromosome 18q21 in the methamphetamine abusers for elucidating the candidate gene for methamphetamine addiction leading to psychosis. We have selected 30 unrelated controls (16 males, 14 females; age=59.8+/-10.4) and 37 male methamphetamine abusers (age=43.3+/-7.8). We analyzed 20 single nucleotide polymorphisms (SNPs) of 7 neuronal genes in chromosome 18q21 for DNA samples that was checked for the data quality and genotype error. The association between the case-control status and each individual SNP was measured using multiple logistic regression models (adjusting for age and sex as covariates). And we controlled false discovery rate (FDR) to deal with multiple testing problem. RESULTS: We found 3 significant SNPs of 2 genes in chromosome 18q21 (p-value<0.05; adjusting for age as covariate) in methamphetamine abusers compared to controls. We also found 2 significant SNPs of 1 gene (p-value<0.05; adjusting for age and sex as covariates) (rs3794899, rs3794901:MAPK4). Two SNPs in MAPK4 gene were significant in both statistical groups. CONCLUSION: MAPK4, the gene for mitogen-activated protein kinase 4, is one of the final 6 candidate genes including ME2 in 18q12-21 in our previous finemapping for psychosis. Our results suggest that MAPK4 can be a candidate gene that contribute to the methamphetamine addiction leading to psychosis.

Effect of clozapine exposed in the pregnant day on the development and the expression of MAPK44/42 in brain of filial rats / 中华行为医学与脑科学杂志

Objective To set up the experimental animal model of the filial rats exposed clozapine in the pregnant clay and expect to know what is the effect on the offspring exposed the antipsychiatrie in gestation.Methods Female Wister rats were divided into both the control group (n=17)and elozapine group (n=2.5) which were intraperitoneally injected saline and clozapine respectively from pregnant day 6 to day 15,and their pup rats were examined development indexes such as the physical growth ,neural reflex,neural behavior and the expression of MAPK44/42 in brain in neonatal stage.Results it was shown that clozapine- exposed in pregnant day lead to the retardation of the weight of filial rats in earlier age such as postnatal day 4 (PND4) and PND7(F=12.56/7.51,P=0.001/0.008).As for neural reflex,the time of cliff avoidance of rats was significantly delayed in PND 3 and day 9 (F=4.969/4.348 ; P=0.03/0.041),and the ratio of positive response of air righting decreased compared with the control group in PND13 and PND15 (F=7.959/6.475,P=0.007/0.016).There was no difference between two groups on the behavior of the filial rats in the open field of filial rats.The expression of PhoMAPK44/42 in hippocampi of exposed-clozapine offspring in PND15 was significantly lower than the control' s(F=18.729/23.824,P=0.001) ,however,no difference was demonstrated on the expression of MAPK44/42 in hippocamaias well as the expression of both Pho-MAPK44/42 and MAPK44/42 in forehead cortex in two groups.Conclusion The resuits suggest the aberration of both physical and mental development of filial rats exposed clozapine in gestation.