ObjectiveTo investigate the effects of ABI-1 gene knockdown upon the proliferation and migration of human gastric cancer cell NCI-N87 in vitro. MethodsNCI-N87-ABI-I-ShRNA cell model was successfully constructed and validated by Real-time PCR and Western blot. The cellular morphous and skeleton, proliferative and migrative potents, and also AKT expression were compared between NCI-N87-ABI-1-ShRNA and its parents by immunofluorental staining, CCK-8 assay, transwell chamber and Western blotting.ResultsCCK-8 assay showed there was no significant difference in the proliferation rates at different time points between the NCI-N87-Vector and NCI-N87 cells while the proliferation rates at the time points of 36 and 48 hours of the NCI-N87-ABI-1-ShRNA were significantly lower than the NCI-N87( t =2. 85and 4. 166, P ＜ 0. 05 ). Transwell assay showed that migrated cell number were 66 ± 8, 65 ± 8 and 30 ± 4,respectively, and there was significant difference between the NCI-N87-ABI-1-ShRNA and NCI-N87 cells (t =9. 550,P ＜0. 05). Finally, ABI-1- knock-down altered the cellular morphoos and skeleton of 90％NCI-N87 cells and inhibited p-AKT expression.ConclusionABI-1 inhibits proliferation and migration of NCI-N87 cells in vitro probably by PI3K/AKT pathway.

Objective To determine whether acute withdrawal of simvastatin treatment in healthy normocholesterolemic men impairs the brachial artery endothelial function.Methods The study was performed on 16 healthy,young male subjects with desirable serum levels of cholesterol.They were administered with simvastatin(20 mg/d)for 4 weeks.Endothelial dependent flow-mediated vasodilation (FMD)was assessed on the brachial artery using high-resolution ultrasound,and fasting serum lipid profiles as well as vasoactive substances[NO,endothelin(ET)and 6-keto-PGF1α] were measured.The parameters mentioned above were obtained at indicated time points before and after simvastatin treatment. Resuts Simvastatin treatment for 4 weeks significantly improved FMD and reduced low density LDL-C and total TC levels.Withdrawal of simvastatin.however,resulted in dramatic impairment of endothelium- dependent relaxation on the first day after with drawal [(4.6±0.48)%and(10.9±0.89)%,P＜0.01 ], Furthermore,FMD decreased significantly as compared with baseline level[(4.6±0.48)%vs(6.4±0.47)%,P＜<0.01].Serum NO level varied according to the change of endothelial-dependent relaxation(γ=0.496,P＜0.01).After discontinuing simvastatin therapy,plasma ET increased and plasma 6-keto-PGF1α decreased progressively.In addition,serum TC and LDL-C were not significantly modified during the initial 2 days.No correlation was shown between FMD and serum LDL-C level(γ=-0.172,P=0.101). Conclusions Withdrawal of simvastatin not only abrogates the beneficial effect on endothelial function of healthy normocholesterolemic men rapidly,but also induces further endothelial deterioration as compared with pretreatment status.This adverse effect is independent of serum cholesterol.The underlying mechanism may be related to the suppression of endothelial NO production.

Objective To study the effect and mechanism of CKLF-like Marvel transmembrane domain containing 5 (CMTMS) on prostate cancer cell proliferation,migration and invasion.Methods The inhibitory effects of CMTM5 on the migration of DU145 cells were studied in vitro by wound healing assay.The expression of the cell signal pathway PI3K-AKT protein was detected by Western blot.The inhibition of tumor growth was also studied in transplanted prostate cancer nude mice model treated with CMTM5 adenovirus.The expression of CMTM5 and ki-67 in transplanted prostate cancer tissue of the nude mice model was analyzed immunohisochemistically.Prostate tumor volume in the nude mice model and the proliferation were measured two weeksafter.injection..Results Wound healing assay showed that over-expression of CMTM5 can inhibit the migration of DU145 cells.The expression of pAKT and NF-kB was significantly decreased after the overexpression of CMTM5.The tumor volume (573.39 ± 175.24) mm3,weight (0.55 ± 0.11) g and proliferation index of prostate in CMTM5 orthotopic injection nude mice model were significantly smaller and decreased than those in the control group (1482.50 ± 327.86) mm3 and (1.31 ± 0.29) g (P ＜ 0.05).Conclusions Both in vitro and in vivo experiments demonstrate that overexpression of CMTM5 could suppress prostate cancer cell proliferation,migration and invasion.The effect may be conducted by PI3K-AKT signaling pathway.

OBJECTIVETo investigate the interaction between myocardial norepinephrine (NET) and protein interacting with kinase Cα (PICK1), and examine the myocardial expression pattern of NET and PICK1 in mice with adriamycin-induced congestive heart failure.

METHODS(1) Cellular experiments: 293T cells were transfected with NET, GFP-PICK1, NET+GFP-PICK1 or NET+GFP-PICK1(KD-AA), respectively. Immunofluorescence staining was performed 48 h after the transfection. (2) Animal experiments: 40 male C57BL/6J mice were divided into control group and adriamycin group (intraperitoneal injection of 2 mg/kg adriamycin with a cumulative amount of 22 mg/kg). The myocardial mRNA and protein expression level of NET, PICK1 and adrenergic receptor (β1-AR) were detected by real-time PCR and Western blot after 10 weeks.

RESULTS(1) PICK1 mediates the intracellular trafficking of NET. (2) Compared to controls, cardiac mRNA expression of NET remained unchanged, but PICK1 and β1-AR mRNA level were significantly reduced in the heart failure mice. (3) Myocardial NET protein expression level was significantly reduced, whereas tyrosine hydroxylase (TH) protein expression was significantly upregulated in heart failure mice. (4) The myocardial density of sympathetic nerve fibers remained unchanged in heart failure mice.

CONCLUSIONSCardiac expression of NET and PICK1 are down-regulated in heart failure mice. Reduced PICK1-mediated intracellular trafficking of NET may be involved in the impairment of NET function in this congestive heart failure mice model.

Animals ; Carrier Proteins ; genetics ; metabolism ; Disease Models, Animal ; Doxorubicin ; adverse effects ; Heart Failure ; chemically induced ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Myocardium ; metabolism ; Norepinephrine Plasma Membrane Transport Proteins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics