PXD101 induces apoptosis of human prostate cancer cell line PC3 by mitochondrial signal pathway / 中国病理生理杂志

AIM:To explore the mitochondrial pathway in the apoptosis of PC3 cells induced by PXD101 (also named as belinostat).METHODS:PC3 cells were treated with PXD101 at different doses or stimulated with PXD101 for different time.The effect of PXD101 on the viability of PC3 cells was measured by CCK-8 assay.The apoptotic rates and the mitochondrial membrane potential (MMP) were analyzed by flow cytometry.The protein levels of Bcl-2, Bax and cytochrome C (Cyt C) were determined by Western blot.The caspase-3 activity were tested by caspase-3 activity assay kit.RESULTS:The viability of the PC3 cells was inhibited by PXD101 in a dose-and time-dependent manner.Flow cytometric analysis showed that the apoptotic rates were increased in the cells treated with PXD101 (P<0.01).At the same time, PXD101 induced the decreases in MMP and Bcl-2, the release of Cyt C, and the increase in caspase-3 activity.CONCLUSION:PXD101 induces the apoptosis of human prostate cancer cell line PC3 by mitochondrial signal pathway.

STC2 promotes proliferation and EMT-related migration of HepG2 cells / 中国病理生理杂志

AIM:To explore the effects of stanniocalcin 2 (STC2) on the proliferation, migration and the process of epithelial-mesenchymal transition (EMT) in human hepatocellular carcinoma HepG2 cells.METHODS:The expression levels of STC2 in the hepatocellular carcinoma cell lines and normal liver cells were assessed by Western blot.Colony formation assay was used to test the effect of STC2 on the proliferation of HepG2 cells.The effects of STC2 on the expression of proliferation-related molecules at mRNA and protein levels were determined by RT-qPCR and Western blot.The effect of STC2 on the migration ability was measured by Transwell assay.The mRNA and protein levels of vimentin and E-cadherin in STC2-overexpressing and-silencing cell lines were detected by RT-qPCR and Western blot.RESULTS:Compared with the normal liver cell line, the protein expression of STC2 was up-regulated in the hepatocellular carcinoma cell lines.The results of colony formation assay indicated that STC2 promoted the proliferation of HepG2 cells.STC2 significantly regulated the proliferation-related gene expression, such as cyclin D1.The results of Transwell assay showed that STC2 enhanced the migration ability of the HepG2 cells and influenced the EMT process.CONCLUSION:STC2 promotes the proliferation of HepG2 cells and affects the expression of proliferation-related genes.STC2 influences the process of EMT and promotes the migration of HepG2 cells.

Screening of plasma biomarkers in patients with unstable angina pectoris with proteomics analysis / 解放军医学杂志

Objective To analyze and compare the differentially expressed plasma proteins between patients with stable angina pectoris (SAP) and unstable angina pectoris (UAP), and search for the biomarkers that maybe used for early diagnosis of UAP. Methods Sixty plasma samples were collected respectively from normal controls group (N group), SAP group and UAP group during Jun. 2014 to Apr. 2015 from the Third Affiliated Hospital of Southern Medical University. Ten samples (100μl) of each group were selected randomly to pool into 3 groups severally. After removing high-abundance proteins from plasma, two-dimensional difference gel electrophoresis (DIGE) was used to isolate the total proteins, and then the protein spots with more than 2-fold changes between UAP and SAP were picked up after the differential software analysis. Afterward, the varied proteins were identified by matrix assisted laser desorption ionization-time of flight/time of flight (MALDI-TOF/TOF) mass spectrometry (MS). Finally, 40 plasma samples were collected respectively from N, SAP and UAP group, and the UAP specific differential proteins were selected to be verified by ELISA. Results A total of 10 varied protein spots with more than 2-fold changes in UAP and SAP were found including 9 up-regulated proteins and 1 down-regulated one. MS identification indicated that the up-regulated proteins included fibrinogen gamma chain (FGG), complement C4-B (C4B), immunoglobulin (Ig) kappa chain C region (IGKC) and hemoglobin subunit alpha (HBA1), whereas the down-regulated one was haptoglobin (HP). After comparing the varied proteins with that in N group, 2 specifically UAP-related proteins, IGKC and HP, were detected totally. IGKC was selected to validate by ELISA, and the corresponding results showed that IGKC was increased specifically in UAP plasma (P<0.05) when compared with N and SAP group, which was consistent with DIGE. Conclusion IGKC and HP have been detected as specifically related proteins to UAP, and IGKC might serve as a potential specific biomarker for screening and early diagnosis of UAP.

Screening of plasma tumor biomarkers for hypopharyngeal carcinoma by proteomics method / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen tumor biomarkers in the plasma close related with hypopharyngeal carcinoma.</p><p><b>METHODS</b>Pooled plasma from 6 patients with hypopharyngeal carcinoma and 6 healthy individuals were collected. After removal of high-abundance plasma proteins, two-dimensional gel electrophoresis (2-DE) was performed to isolate the total proteins, and the protein spots with more than 2-fold differential expressions were detected by 2D analysis software followed by identification by MALDI-TOF/TOF mass spectrometer. Western blotting was performed to validate the expression level of α2-HS-glycoprotein.</p><p><b>RESULTS</b>A total of 11 differentially expressed protein spots were selected, including 5 upregulated proteins and 6 downregulated proteins. MALDI-TOF/TOF identified the upregulated proteins in hypopharyngeal carcinoma patients as alpha-2-HS-glycoprotein and haptoglobin and downregulated ones as Ig kappa chain C region and apolipoprotein A-I. Western blotting demonstrated that α-2-HS- glycoprotein expression level was consistent with that detected by 2-DE.</p><p><b>CONCLUSION</b>Patients with hypopharyngeal carcinoma show different plasma protein profiles from healthy individuals. These differentially expressed proteins may serve as potential specific tumor biomarkers for hypopharyngeal carcinoma.</p>

Screening of differential proteins binding to Nox1 promoter in A549 cell model of inflammation and oxidative stress / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen the regulatory proteins involved in Nox1 promoter activation in a cell model of inflammation and oxidative stress.</p><p><b>METHODS</b>A cell model of inflammation and oxidative stress was established by stimulating A549 cells with tumor necrosis factor-α (TNF-α). The differential proteins binding to Nox1 promoter were screened by DNA pull-down and the binding proteins were separated by 2D electrophoresis and selected according to the their differential expression levels (with over 1.5-fold changes relative to the control level). The screened proteins were finally identified by MALDI-TOF/TOF-MS.</p><p><b>RESULTS</b>Seven differentially expressed protein spots (all upregulated in the cell model) were obtained, among which GLE1, DDX19A, KRT1 and KRT10 were identified by mass spectrometry.</p><p><b>CONCLUSION</b>GLE1, DDX19A, KRT1 and KRT10 participate in the activation of Nox1 promoter in TNF-α-induced A549 cells, and this result provides new insights into the biological roles of the regulatory proteins of Nox1 promoter in inflammation and oxidative stress.</p>

Screening and mass spectrometry analysis of differentially expressed proteins of plasm between laryngocarcinoma and healthy individuals / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To compare and analyze the expression difference of proteins between laryngocarcinoma and healthy individuals to search for protein biomarkers that may be detected in plasm of laryngocarcinoma patients.@*METHOD@#Pooled plasm from 6 laryngocarcinoma patients and 6 healthy individuals as controls were collected. Two-dimensional gel electrophoresis (2-DE) was used to isolate the total proteins, and the differential protein spots were identified by MALDI-TOF/TOF mass spectrometer, and then the biological information of the proteins was analyzed.@*RESULT@#Twenty differential expressed protein spots with more than 1.5-fold between the laryngocarcinoma and healthy individuals were selected, and there were 8 proteins upregulated and 12 proteins downregulated among these proteins. After identifying by MALDI-TOF/TOF, compared with healthy controls, the spots that were L1, C-reactive protein, haptoglobin, ceruloplasmin; the spots that were downregulated were: Serotransferrin, alpha-2-HS-glycoprotein, fibrinogen gamma chain, haptoglobin related protein, Ig lambda-1 chain C regions, Ig kappa chain C region, apolipoprotein A-I, transthyretin, apolipoprotein C-III.@*CONCLUSION@#Compared with laryngocarcinoma and healthy individuals, the plasm proteins profile showed differently. The proteins of differential expressed are expected to be the specific plasm biomarkers for laryngocarcinoma patients.