MIA3 promotes the degradation of GSH (glutathione) by binding to CHAC1, thereby promoting the progression of hepatocellular carcinoma.

MIA3 (melanoma inhibitory active protein 3)/TANGO1 (Golgi transporter component protein) plays an important role in the initiation, development, and metabolism of cancer. We aimed to explore the role and underlying molecular mechanisms of MIA3/TANGO1 in the growth and migration of hepatoma cells. According to the analysis of The Cancer Genome Atlas (TCGA) database, MIA3 is expressed at higher levels in hepatocellular carcinoma (HCC) tissues than in normal tissues. Real-time quantitative polymerase chain reaction (qRT-PCR), immunohistochemistry, and western blotting were used to detect mRNA and protein expression in HCC tissues and cells. The in vitro function of MIA3 in HCC cells was evaluated using Cell Counting Kit-8 (CCK-8), colony formation, cell migration and invasion, and flow cytometry assays. Hep-G2 cells with MIA3 overexpression were subjected to RNA-seq, and the downstream target gene CHAC1 (glutathione-specific γ-glutamyl cyclotransferase 1) was selected according to the results of the volcano map of gene enrichment. The relationship between MIA3 and CHAC1 was revealed by coimmunoprecipitation and confocal microscopy. MIA3 expression was upregulated in HCC organizations and HCC samples in the TCGA dataset. Knocking out MIA3 inhibited the proliferation, migration, and invasion of Hep-G2 cells and promoted the apoptosis of Hep-G2 cells. Overexpression of MIA3 in Huh7 cells promoted the proliferation, migration, and invasion and suppressed the apoptosis of Huh7 cells. Overexpression of MIA3 promoted the expression of CHAC1 and the degradation of glutathione (GSH), thereby promoting the growth and metastasis of HCC cells. Knocking out MIA3 inhibited the expression of CHAC1 and slowed the degradation of glutathione, thereby inhibiting the growth and metastasis of HCC cells. MIA3 further promotes the growth, metastasis, and invasion of hepatoma cells by binding to the CHAC1 protein and promoting GSH degradation.

Study on the Effects of Deoxyschizandrin on the Proliferation ，Migration and Invasion of Human Nasopharyngeal Carcinoma Cell HONE- 1 via Met/PI 3K/Akt Signaling Pathway / 中国药房

OBJECTIVE：To study the effects and potential mechani sm of deoxyschizandrin on the proliferation ，migration and invasion of nasopharyngeal carcinoma cell HONE- 1. METHODS ：HONE-1 cell was set as cell model ，while CCK- 8 test，wound healing assay and Transwell chamber test were used to detect the proliferation ，migration and invasion ability changes of HONE- 1 cells after treatment with different concentrations [ 0（blank control ），10，20，40 μmol/L] of deoxyschizandrin. Computer molecular docking was performed to analyze the binding ability between deoxyschizandrin and Met protein. Western blotting assay was used to detect the relative protein expressions of p-Met ，p-PI3K，p-Akt，Bcl-2 and N-cadherin in cells. RESULTS ：Compared with blank control ，the proliferation ，migration and invasion ability of cells after treated with 10，20，40 μmol/L deoxyschizandrin were all decreased significantly （P＜0.05）. Results of molecular docking revealed that deoxyschizandrin could stably bind with the activity pocket of Met protein. Results of Western blotting assay demonstrated that compared with blank control ，10，20，40 μmol/L deoxyschizandrin all decreased the relative protein expressions of p-Met ，p-PI3K，p-Akt，Bcl-2 and N-cadherin in cells significantly（P＜0.05）. CONCLUSIONS ：Deoxyschizandrin can inhibit the proliferation ，migration and invasion of HONE- 1 cell via inhibiting the activation of Met/PI 3K/Akt signaling pathway.

Levels of histone modifications in activated primarily cultured rat hepatic stellate cells / 中国病理生理杂志

AIM:To investigate the changes of histone modifications during the activation of primarily cultured rat hepatic stellate cells ( HSCs) and the relationship between histone modification patterns andα-smooth muscle actin (α-SMA) expression, and to explore the roles of histone modifications in the activation of HSCs.METHODS:The rat HSCs were isolated by in situ perfusion of collagenase combined with density gradient centrifugation, cultured in vitro and identi-fied by immunofluorescence staining.The morphological features of the cells were observed under inverted microscope.The changes of desmin and α-SMA during the activation of HSCs were detected by immunofluorescence staining and Western blotting.The levels of histone 3 lysine 4 dimethylation (H3K4me2), histone 3 lysine 9 dimethylation (H3K9me2), his-tone 3 lysine 9 acetylation (acH3K9) and histone 4 lysine 12 acetylation (acH4K12) in quiescent HSCs and activated HSCs were determined by Western blotting.RESULTS: The morphology of HSCs shifted from a quiescent phenotype to highly activated myofibroblast during the culture.Immunofluorescence staining and Western blotting showed that the expres-sion levels of α-SMA and desmin were increased over time and reached maximum at 15 d.According to the results of cell morphology and immunofluorescence staining, the cells cultured for 24 h and 15 d were quiescent and activated HSCs, re-spectively.Compared with quiescent HSCs, there were higher H3K4me2 and lower H3K9me2, acH3K9 and acH4K12 modification levels in activated HSCs ( P<0.01 ) .CONCLUSION: Histone modifications show anomalous expression during the activation of primarily cultured rat HSCs.Histone modifications may contribute to the transdifferentiation of HSCs and the development of hepatic fibrosis.

Effects of sinomenine and acupuncture on amygdala in heroin-withdrawal rats / 中国药理学通报

Aim To explore the effects of acupuncture and sinomenine on the withdrawal symptoms,on the spontaneous discharge and synaptophysin expression of amygdala of heroin-addicted rats.Methods 75 SD rats were randomly and equally divided into 5 groups consisted of control,heroin-addicted,acupuncture,sinomenine and combined treatment group.The model of heroin-addicted rats were established with heroin subcutaneous injection,which in acupuncture,sinomenine and combined group were subsequently punctured at 3 acupoint(Neiguan,Baihui and Zusanli),intramuscular injected with sinomenine and treated with combined methods(acupuncture and sinomenine)respectively.The withdrawal symptoms of rats in different group were evaluated,and then the spontaneous discharges and synaptophysin expression of amygdala were detected with electrophysiological and immunohistochemical technology respectively.Results The heroin-addicted rats were successfully established,being suggested by typical withdrawal symptoms induced by naloxone(P<0.01 vs control),which were alleviated in the 3 treatment groups(acupuncture,sinomenine and combined ) with varying degree(P<0.01 vs heroin group).The electrophysiological results suggest that the forms of low-frequency and single irregular discharges in amygdala were increase,bunchy/cluster and complex discharges were decreased(P<0.05 vs control),which were reversed apparently in sinomenine and combined treatment groups(P<0.05 vs heroin group),closed to control.The immunohistochemical results show that expression of synaptophysin in amygdala was increased obviously(P<0.01 vs control),which was decreased in the 3 treatment groups(acupuncture,sinomenine and combined),P<0.01 vs addicted group.Conclusion Treatments of acupuncture and sinomenine administration can alleviate withdrawal symptoms of heroin-addicted,and rectify the abnormal spontaneous discharges and expression of synaptophysin in amygdala of heroin-addicted rats for certain degree,especially with combined treatments.

Two dimensional liquid phase chromatographic fractionation of phosphoproteome of mouse liver / 解放军医学杂志

Objective To fractionate phosphoproteome of mouse liver by two-dimensional (2D) liquid phase chromatography fractionation. Methods Phosphoproteins were extracted from lysates of normal mice livers by phosphate metal affinity chromatography (PMAC) resin. The phosphoproteins were exchanged by start buffer and separated by chromatofocusing in the first dimension. Then the fractions between pH 8.5 and pH 4.0 were separated by non-porous silica (NPS) reverse-phase high performance liquid chromatography (RP-HPLC). Finally, the UV maps were converted into gel-like maps by ProteoVue software. Results Phosphoproteins of mouse liver were successfully extracted and fractionated by two dimensional liquid phase chromatographic fractionation after concentration and desalt. Then pI/UV map of mouse liver phosphoproteome was successfully set-up. There are 16 fractions between pH 8.5 and pH 4.0 after chromatofocusing in the first dimension and the UV maps of each fraction were converted into pI/UV gel-like maps. Conclusions Combination of technique of phosphoproteins enrichment and 2-D liquid phase chromatographic fractionation is an effective approach to research phosphoproteome and the key base for further identification and investigation of phosphoproteins.