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Objective@#To investigate the expressions of extracellular signal-regulated kinase (ERK) and p-ERK in benign and malignant prostate tissues, and whether it can be used as a marker for the prognosis of advanced prostate cancer (PCa).@*METHODS@#Using immunohistochemical Envision, we detected the expressions of ERK1/2 and p-ERK1/2 in 20 cases of benign prostatic hyperplasia (BPH) and 40 cases of advanced PCa and analyzed their correlation with PCa metastasis, Gleason score, PSA level, and prognosis.@*RESULTS@#The expression of ERK1/2 was remarkably higher in the advanced PCa than in the BPH cases (82.5% vs 55%, P5 yr, and survival ≤ 5 yr groups were 61.9%, 89.5%, 57.9%, and 90.5%, respectively, with statistically significant differences among these groups (P<0.05).@*CONCLUSIONS@#ERK1/2 and p-ERK1/2 proteins are highly expressed in advanced PCa and p-ERK1/2 is associated with the metastasis and prognosis of advanced PCa.
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Humans , Male , Biomarkers, Tumor , Metabolism , Extracellular Signal-Regulated MAP Kinases , Metabolism , Mitogen-Activated Protein Kinase 1 , Metabolism , Mitogen-Activated Protein Kinase 3 , Metabolism , Neoplasm Grading , Neoplasm Metastasis , Prognosis , Prostate , Prostate-Specific Antigen , Metabolism , Prostatic Hyperplasia , Pathology , Prostatic Neoplasms , Mortality , PathologyABSTRACT
ObjectiveTo study the effect of C3G gene on apoptosis and proliferation of H9C2 cardiomyocytes and its mechanism. MethodsThe RNA lentivirus was constructed, and pCXN2-Flag plasmid (empty plasmid) and pCXN2-Flag-hC3G plasmid (over-expressed human C3G mRNA) were purchased. H9C2 cardiomyocytes were respectively infected and transfected with blank reagent, negative lentivirus, C3G siRNA lentivirus, C3G siRNA lentivirus+pCXN2-Flag plasmid and C3G siRNA lentivirus+pCXN2-Flag-hC3G plasmid with stochastic method. Thus the experiments were randomly divided into five groups, namely blank group, negative control group, C3G silence group, C3G silence+empty plasmid group, and C3G silence+C3G overexpression group. Seventy two hours after plasmid transfection, the expression of C3G mRNA was detected by PT-RCR, cell apoptosis was determined by flow cytometry, cell proliferation rate was determined by MTT, and the protein levels of C3G, p-ERK1/2, Bax and Flag were determined by Western blotting. Results As screened by puromycin, it was found that more than 85% of the cells in negative control group and C3G silence group were labeled with green fluorescent protein, showing that more than 85% of the cells were infected with lentivirus. Compared with blank group and negative control group, the expressed Bax protein and cell apoptosis rate were increased significantly (P<0.01, P<0.05), and the expression levels of C3G mRNA, C3G protein and p-ERK1/2 protein and the cell proliferation rate were remarkably decreased in C3G silence group and C3G silence+empty plasmid group (P<0.01, P<0.05); Compared with C3G silence group and C3G silence+empty plasmid group, the expression level of Bax protein and the cell apoptosis rate were decreased obviously (P<0.01, P<0.05), while the expression levels of C3G mRNA, C3G protein and p-ERK1/2 protein and the cell proliferation rate were remarkably increased in C3G silence+C3G overexpression group (P<0.01, P<0.05). Conclusion C3G gene silence can induce apoptosis and inhibit proliferation in H9C2 cardiomyocytes, and overexpression of C3G gene can reverse the effects of C3G gene silence affecting in H9C2 cardiomyocytes, characterized by a reduction of apoptosis rate and promotion of proliferation, and they may be related to p-ERK1/2 protein and pro-apoptotic molecule Bax.
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This study was aimed to investigate the effects of ursolic acid on human hepatic stellate cells (HSC-LX-2) proliferation and its mechanism.Different doses of ursolic acid were incubated with HSC-LX-2 cellin vitrof or 48 h.MTT was used for the detection of HSC-LX-2 cell proliferation.The expressions of PDGF-ERK signaling pathway associated proteins were measured by western blot.The results showed that the proliferation of HSC- LX-2 cells was inhibited by ursolic acid in a dose-dependent manner.The inhibition rate of 20,30 and 40μmol·L-1 of ursolic acid was 9.1%,42.3% and 62.6%,respectively.The IC50 was 35.2μmol·L-1.After incubated with ursolic acid for 48 h,protein levels of PDGF-R and p-ERK in 30 and 40μmol·L-1 group were significantly decreased when compared with the normal group (P<0.05 orP<0.01),except the ERK protein.It was concluded that ursolic acid can inhibit HSC-LX-2 cell proliferation.Its mechanism may be related to the blockage of PDGF-ERK signaling pathway.
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Objective To investigate degenerative changes of auditory functions and age -related expression of phosphorylated extracellular signal -regulated protein kinases1/2(p-ERK1/2) in the cochlea of the senescence accelerated mouse .Methods The 3 ,5 ,7 months old mice were selected for the study and each group had 6 mice . The 8 kHz tone burst auditory thresholds and age -related expression of p -ERK1/2 in the cochlea were studied in the senescence accelerated mouse/prone 8(SAMP8) at 3 ,5 ,7 months .The expression of p -ERK1/2 was analyzed by the optical density of immunohistochemical staining .Results For the auditory function evaluation :The SAMP 8 developed a progressive hearing loss at 8 kHz which showed an age -related significant increase (P< 0 .05) .The ABR thresholds in 3 ,5 ,7 months old groups in the left ear were 31 .817 ± 1 .228 ,54 .329 ± 1 .459 ,58 .330 ± 1 .252 dB SPL ,respectively .In the right ears ,the ABR threshold were 32 .474 ± 1 .041 ,53 .485 ± 1 .385 ,57 .842 ± 1 .173 dB SPL ,respectively .p-ERK1/2 protein expressed in the cochlea of the SAMP8 throughout the development sta‐ges ,which developed an age-related significant decrease (P<0 .05) .The average optical density of p -ERK1/2 in the spiral ganglion cells in the 3 ,5 ,7 months old mice were 0 .699 7 ± 0 .018 8 ,0 .621 5 ± 0 .014 7 ,0 .575 3 ± 0 .015 5 ,respectively .In the hair cells ,they w ere 0 .651 9 ± 0 .025 2 ,0 .591 2 ± 0 .010 2 ,0 .559 3 ± 0 .006 7 respec‐tively .Conclusion The expression level of p -ERK1/2 protein decreases when the SAMP 8 develops a progressive hearing loss .This indicates that p-ERK1/2 protein probably has relationship with maintaining functional status of the cochlea and the auditory formation .
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Objective To mvestigate the effect of phospholated-ERKl/2 on NF-κB p65 expresson nn the substania nigra(SN) of l-msthyi-4-phenyi-l,2, 3, 6-tetrahydropyeidine (MPTP)-induced mouse model of Parkinson's disease(PD). Methods PD mouse model was induced by MPTP and the behavoor of mouse was observed. Immunohistochemistry and Western bloiting analysis were used to observe the changes in expression of tyrosine hydroxylase (TH), NF-κB p65 and p-ERKl/2 in the SN of midbrain. Meanwhite, the above changes were also observed after treatment with U0126, a specific inhibitor of ERK. Results 1 h after the third MPTP administration, there were much more p-ERKl/2 positive cells than NF-κB p65 positive cells in the SN. 24 h after the fifth injection of MPTP, NF-κB p65 positive cells were significantly increased and p-ERKl/2 positive celiswere decreased, accompanted by marked loss of TH positive neurons. The above changes were greatly alleviated in animais treated with U0l26. Conclusion ERKl/2 pathway may regulate NF-κB p65 activation in MPTP-induced mouse model of Parkinson's disease, which leads to loss of dopamine neurons.
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AIM:To investigate the effect of ERK1/2/c-Fos signal pathway during angiotensin-(1-7)inhibiting proliferation of rat glomerular mesangial cell strain(GMCS)induced by angiotensin Ⅱ.METHODS:Rat glomerular mesangial cells(GMC)were co-cultured with angiotensin Ⅱ and different doses of angiotensin-(1-7).The numbers of GMC were evaluated by crystal violet staining.The amounts of p-ERK1/2 and c-Fos expressions were detected by Western blotting.RESULTS:Angiotensin-(1-7)showed its inhibitory effects on GMC number increasing induced by angiotensin Ⅱ as well as the amounts of p-ERK1/2 and c-Fos expressions in a concentration dependent manner.CONCLUSION:ERK/c-Fos signal pathway is involved in the inhibitory effects of angiotensin-(1-7)on angiotensin Ⅱ-induced GMC proliferation.