Expression and distribution of C1C-3 in human glioma specimen

AIM: To investigate the expression of voltage-gated chloride channels (ClC)-3 protein and mRNA in human glioma specimen and its biological function. METHODS: The expression of C1C-3 was observed by immunohistochemical staining in 24 cases of human glioma, 4 cases of brain metastic cancer specimens and 5 cases of normal brain tissue as control; The C1C-3 mRNA expression were detected in the specimens with positive expression of ClC-3 protein by RT-PCR. RESULTS: ClC-3 protein was found negative in 4 cases of normal brain tissues and positive in 19 cases of human glioma and 4 cases of brain metastic cancer specimens. ClC-3 protein was mainly expressed in the membrane or cytoplasm of neoplastic cells and microvascular endothelial cells. The expression of ClC-3 mRNA was detected in 16 cases of human glioma and 4 cases of brain metastasis cancer specimens among the tissues with the positive expression of ClC-3 protein. The level of protein and RNA of ClC-3 in high malignant oligodendrogliomas was higher than that in low malignant ones. CONCLUSION: ClC-3 is generally expressed in human glioma and brain metastic cancer and is probably correlated with the classification of its pathological malignance.     

Modulation of bacterial redox protein azurin induces human osteosarcoma cell apoptosis by Fas antigen and caspase-8

AIM: To determine the role of Fas antigen and caspase-8 in modulating apoptosis of osteosarcoma cells induced by bacterial redox protein azurin. METHODS: The human osteosarcoma cell line U2OS was treated with bacterial redox protein azurin (150 mg/L) for 6, 12, 24 and 48 h, respectively. Cell immunohistochemistry and quantitative image pattern analysis were applied for detecting the expression of Fas antigen. Caspase-8 activity was detected using caspase-8 fluorescent assay kit. The apoptotic rate was measured by FCM. RESULTS: Compared with the control group, the expression of Fas antigen and activity of caspase-8 significantly increased in U2OS cells treated with 150 mg/L azurin (P<0.01). There was positive correlation between the expression of Fas antigen and activity of caspase-8 (r=0.873, P<0.01). The increased Fas antigen expression had the function to transfer apoptotic signals and the anti-Fas antibody promoted azurin induced apoptosis through increasing Fas antigen expression. IETD-FMK blocked the activation of procaspase-8 and reduced apoptosis of U2OS cells in the presence of azurin or anti-Fas antibody. The apoptotic rates in azurin group and anti-Fas antibody group were significantly different from the inhibitor group (P<0.01). CONCLUSION: The Fas-dependent signalling is the important pathway of azurin inducing apoptosis in U2OS cells. The up-regulation of csepase-8 may play an important role.     

Expression of SAL-like 4 protein in human prostate cancer tissue/cell lines and its relationship with clinical outcome

AIM:To evaluate the expression of SAL-like 4 (SALL4) protein in human prostate cancer cell lines and tissues, and to analyze the relationship between SALL4 expression and the clinicopathological parameters. METHODS:Immunofluorescence, RT-PCR and Western blotting were performed to detect the expression of SALL4 at mRNA and protein levels in 3 common prostate cancer cell lines LNCaP, DU145 and PC-3. The normal prostate epithelial cell line RWPE-1 was used for control. The protein levels of SALL4 in the tissues of benign prostate hyperplasia and prostate cancer tissues were determined by the method of immunohistochemistry. RESULTS:The SALL4 protein was predominantly expressed in the cytoplasm of the cells. The protein levels of SALL4 in 3 common prostate cancer cell lines were significantly higher than that in RWPE-1 cells. However, the mRNA level of SALL4 had no obvious difference among the 4 cell lines. Immunohistochemistry results showed that the expression level of SALL4 in the cancerous tissues was significantly higher than that in noncancerous (benign and normal) prostatic tissues. In addition, we found that the expression level of SALL4 in prostate cancer was significantly correlated with the Gleason score, clinical stage, prognosis estimation and tissue prostate-specific antigen (PSA) expression, but not associated with age, the level of serum total PSA, prostate volume and the expression of androgen receptor in the tissues of the patients. CONCLUSION: The over-expression of SALL4 protein may play an important role in the pathogenesis and progression of prostate cancer, and provides some reference indexes for estimating the malignancy, progression and prognosis of prostate cancer.     

Effect of TAP-SSL5 fusion protein on binding of activated platelets to human lymphocytes

AIM: To study the effect of tick anticoagulant peptide-staphylococcal superantigen like protein 5 (TAP-SSL5), an anti-inflammatory and anticoagulant fusion protein, on the binding of activated platelets to human lymphocytes.METHODS: Human periphery lymphocytes were isolated by magnetic activated cell sorting (MACS). The toxicity of TAP-SSL5 on the viability of Jurkat cell was assessed by CCK-8 assay. Flow cytometry was applied to detect the expression of CD162 (PSGL-1) on the Jurkat cells (human peripheral blood leukemia T lymphocyte cell line) and the inhibitory effect of TAP-SSL5 on the binding of mouse anti-human CD162 monoclonal antibody (KPL-1) to Jurkat cells. Platelets were activated by ADP at concentration of 20 μmol/L, the binding rates of activated platelets to Jurkat cells or human lymphocytes were assayed by flow cytometry. RESULTS: The concentration of TAP-SSL5 below 30 mg/L didn't affect the viability of Jurkat cells. TAP-SSL5 at 10 mg/L competitively inhibited KPL-1 binding to Jurkat cells. The binding rates of activated platelets to Jurkat cells or lymphocytes were (11.86±4.49)% and (8.32±1.00)%, respectively, which decreased to (6.73±2.71)% and (5.51±0.70)% after the Jurkat cells and lymphocytes were pre-incubated with 10 mg/L TAP-SSL5 (P <0.05).CONCLUSION: TAP-SSL5 binds to PSGL-1 expressed on lymphocyte surface and directly inhibits the binding of activated platelets to human lymphocytes, which may be one of the anti-inflammatory mechanisms of TAP-SSL5.     

Effect of down-regulation of X-box binding protein 1 gene expression on viability and apoptosis of glioma cells

AIM: To investigate the effect of down-regulation of X-box binding protein 1 (XBP1) expression on the viability and apoptosis of glioma cells. METHODS: The mRNA expression of XBP1 in the glioma tissues was detected by qPCR. Small interfering RNA (siRNA) interfering with XBP1 expression (XBP1-siRNA) was transfected into human brain glioma U251 cells. At the same time, control group (the cells without special treatment) and negative control (NC-siRNA) group (transfected with siRNA without any interference) were set up. The mRNA expression of XBP1 in the 3 groups 48 h after transfection was detected by qPCR. The protein levels of XBP1, proliferating cell nuclear antigen (PCNA), B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2-associated X protein (Bax), cyclin D1 (cyclin D1), phosphatidylinositol 3-kinase (PI3K) and phosphorylated Akt (p-Akt) were determined by Western blot. The cell viability was measured by CCK-8 assay. The cell cycle distribution and apoptosis were analyzed by flow cytometry. RESULTS: The expression level of XBP1 in the glioma tissues was significantly higher than that in the tumor adjacent tissues (P<0.05). The XBP1 expression at mRNA and protein levels was significantly decreased in the cells transfected with XBP1-siRNA (P<0.05). No statistically significant difference of the cell viability, cell cycle, apoptotic rate and the protein levels of PCNA, Bcl-2, Bax, cyclin D1, PI3K and p-Akt between NC-siRNA group and control group was observed. Compared with control group, the cell viability, S-phase cells and the protein levels of PCNA, Bcl-2, cyclin D1, PI3K, and p-Akt in XBP1-siRNA group were decreased significantly, and the apoptotic rate, G₀/G₁-phase cells and Bax protein expression were significantly increased (P<0.05). CONCLUSION: Down-regulation of XBP1 gene expression in brain glioma cells reduces the viability of cancer cells, blocks the cells in G₁ phase and promote apoptosis. The mechanism is related to the inhibition of PI3K/Akt signaling pathway.     

Effects of IGFBPrP1 and thioacetamide on liver tissues in mice

AIM: To investigate the effects of insulin-like growth factor binding protein related protein 1(IGFBPrP1) and thioacetamide (TAA) on the liver tissues, and to identify the role of IGFBPrP1 in liver fibrosis. METHODS: Thirty-two male C57BL/6 wild-type mice were randomly divided into 4 groups (n=8 in each group): control group, recombinant murine IGFBPrP1(rmIGFBPrP1) 4 weeks group, TAA 2 weeks group and TAA 4 weeks group. The methods of hematoxylin-eosin (HE) staining, picric acid-Sirius red staining, immunohistochemistry and Western blotting were performed. RESULTS: The extensive fatty degeneration of liver cells in rmIGFBPrP1 4 weeks group was observed. The collagen deposition was found in TAA 2 weeks group. In TAA 4 weeks group, the degree of hepatic fibrosis was more serious than that in TAA 2 weeks group. The expression levels of IGFBPrP1, transforming growth factor beta 1(TGF-β₁), Smad3, p-Smad2/3, collagen Ⅲ, collagenⅠand fibronectin (FN) in liver tissues were higher in rmIGFBPrP1 4 weeks group, TAA 2 weeks group and TAA 4 weeks group than those in control group. No significant difference of the expression levels of IGFBPrP1, collagen I and FN between rmIGFBPrP1 4 weeks group and TAA 2 weeks group was observed. CONCLUSION: IGFBPrP1 plays an important role in the process of thioacetamide-induced liver fibrosis. Meanwhile, IGFBPrP1 induces excessive deposition of extracellular matrix through TGF-β₁/Smad3 pathway.     

ACTL8 expression and its correlation with clinicopathological features and prognosis in breast cancer

AIM: To investigate the actin-like protein 8 (ACTL8) expression and its relationship with clinicopathological features and prognosis in breast cancer.METHODS: The expression of ACTL8 in human normal mammary epithelial cell line MCF-10A and 5 breast cancer cell lines was detected by Western blot. The expression of ACTL8 was also investigated by immunohistochemistry in 6 cases of breast cancer specimens with adjacent normal tissues. The data in 488 cases of breast specimens from TCGA dataset were downloaded, and the relationship between the mRNA expression of ACTL8 and the clinicopathological features and prognosis was analyzed.RESULTS: The expression of ACTL8 in 4 breast cancer cell lines was significantly higher than that in breast epithelial cell line MCF-10A.The level of the ACTL8 expression in breast tumors was significantly higher than that in the corresponding adjacent normal breast tissues. The mRNA expression of ACTL8 was correlated with age, tumor size, clinical TNM stage and lymph node metastasis of breast cancer patients (P < 0.05). The high expression level of ACTL8 mRNA indicated a poor prognosis of breast cancer patients. CONCLUSION: ACTL8 protein is highly expressed in breast cancer specimens and is closely correlated with the clinicopathological features and prognosis, suggesting that ACTL8 is a prognostic marker for breast cancer or a potential new target for treatment of breast cancer.     

EGFR-p38 MAPK signaling pathway is involved in expression of HMGB1 in pulmonary tissues of rats with ventilator-induced lung injury

AIM: To investigate the role of epidermal growth factor receptor (EGFR)-p38 mitogen-activated protein kinase (MAPK) pathway in the expression of high mobility group box 1 protein (HMGB1) in the lung tissues of rats with ventilator-induced lung injury (VILI).METHODS: Thirty-two healthy Sprague-Dawley (SD) rats were randomly divided into 4 groups (n=8 each): group A, spontaneous breathing; group B, small tidal volume ventilation (VT=8 mL/kg); group C, high tidal volume ventilation (VT=40 mL/kg); group D, high tidal volume ventilation plus EGFR antagonist AG-1478. The rats in group B, group C and group D were mechanically ventilated for 4 h and then all animals were sacrificed.Total protein content and white blood cell (WBC) count in bronchoalveolar lavage fluid (BALF), the lung wet/dry weight ratio (W/D) and myeloperoxidase (MPO) activity were determined. The histological changes of lung tissues were observed by HE staining. The EGFR protein and mRNA expression, p38 MAPK activity and HMGB1 protein expression in the lung tissues were also detected.RESULTS: The inflammatory responses as evidenced by lung HE staining, total protein and WBC in BALF, the lung W/D and MPO activity were significantly higher in group C than those in group A (P<0.05). The mRNA expression of EGFR, EGFR activity, p38 activity and HMGB1 protein level also significantly increased in group C (P<0.05) as compared with group A. Significant decreases in the above indexes in group D were observed as compared with group C.CONCLUSION: High tidal volume ventilation induces acute lung injury, which may be related to up-regulation of HMGB1 expression through EGFR-p38 MAPK signal pathway.     

Expression and clinical significance of bone morphogenetic protein 3 in hilar cholangiocarcinoma tissues

AIM: To investigate the expression and clinical significance of bone morphogenetic protein 3 (BMP3) in hilar cholangiocarcinoma tissues.METHODS: Thirty cases of hilar cholangiocarcinoma specimens were collected. The expression of BMP3 at mRNA and protein levels in the tumor tissues and paracancerous tissues was detected by real-time PCR and Western blot. The hilar cholangiocarcinoma paraffin-embedded specimens (n=103) were collected. The protein expression of BMP3 was determined by immunohistochemical method, and the relationship of BMP3 protein expression with clinical pathological characteristics was evaluated.RESULTS: In the 30 patients with hilar cholangiocarcinoma, the expressions of BMP3 protein and mRNA in 22 cases of tumor tissues were significantly decreased compared with the adjacent normal tissues. The results of immunohistochemistry showed that 87 cases were negative and 16 cases were weakly positive in all 103 cases of hilar cholangiocarcinoma. The expression of BMP3 protein was associated with the tumor TNM staging, lymph node metastasis and tumor differentiation (P<0.05).CONCLUSION: BMP3 gene might be inhibited in human hilar cholangiocarcinoma. The down-regulation of BMP3 gene might be associated with the carcinogenesis and development of hilar cholangiocarcinoma.     

Effect of P53 binding site on regulation of human NOD8 gene

AIM: To investigated the characterization and biological function of P53 binding element in the regulation of NOD8 gene. METHODS: Using the method of bioinformatics, we found a completely preserved P53 binding site in NOD8 core promoter both in humans and chimpanzees. NOD8 gene was amplified by PCR from human cDNA and correctly connected into the vector p NOD8 (760 bp)-EGFP-C2/ mp NOD8 (750 bp)-EGFP-C2. The constructed plasmids p NOD8 (760 bp)-EGFP- NOD8 and mp NOD8 (750 bp)-EGFP- NOD8 were transiently transfected into the cell line HEK293 by JetPei^TM and treated with pifithrin alpha (PFT-α,an inhibitor of P53) at different concentrations for 24 h. The mRNA and protein expression levels of NOD8 were detected by RT-PCR and Western blotting. In addition, chromatin immunoprecipitation (ChIP) assay was performed to determine the binding of P53 to the NOD8 promoter after recombinant plasmid p NOD8 (760 bp)-EGFP was transfected into HEK293 cells. RESULTS: The plasmids p NOD8 (760 bp) -EGFP- NOD8 and mp NOD8 (750 bp)-EGFP- NOD8 were successfully constructed and conformed by restriction digestion and sequence analysis. The results of ChIP confirmed that P53 bound to the promoter of NOD8 . The mRNA expression of NOD8 in the cells transfected with p NOD8 (760 bp)-EGFP- NOD8 was stronger than that in the cells transfected with control vector pEGFP-C2 (P<0.05). Furthermore, the mRNA expression of NOD8 was reduced in HEK293 cells transfectnt with the mutant plasmid mp NOD8 (750 bp)-EGFP- NOD8 compared with the cells transfected with p NOD8 (760 bp)-EGFP- NOD8 . Meanwhile, PFT-α inhibited the mRNA expression of NOD8 in the cells transfected with p NOD8 (760 bp)-EGFP- NOD8 in a concentration-dependent manner, and PFT-α at concentration of 90 μmol/L was the most effective in inhibiting NOD8 mRNA expression (P<0.01). As expected, the protein expression of NOD8 in pNOD8 (760 bp)-EGFP- NOD8 group significantly increased compared with that in pNOD8 -C2 group, the protein expression of NOD8 in mp NOD8 (750 bp)-EGFP- NOD8 group or pNOD8 (760 bp)-EGFP- NOD8 + PFT-α group was dramatically decreased compared with that in p NOD8 (760 bp) -EGFP- NOD8 group. CONCLUSION: The results suggest that the P53 binding site is critical for the regulation of NOD8 gene and there is positive feedback regulation between P53 binding site and NOD8 , which may maintain efficient balance between defense and self-inflicted injury in response to the invasion of pathogen.     

Role of GRP78 in alteration of myocardium induced by intestinal endotoxemia in cirrhotic rats

AIM: To explore the role of glucose-regulated protein 78 (GRP78) in the alteration of myocardium induced by intestinal endotoxemia in cirrhotic rats. METHODS: Fifty-one male Wistar rats were randomly divided into liver cirrhosis groups of 4-week, 6-week and 8-week, and normal control groups at corresponding time points. The cardiac functions of the 8-week rats were measured. Tumor necrosis factor α(TNF-α) and malondialdehyde(MDA) in myocardial tissues were detected. The number of myocardial cells and the collagen volume fraction (CVF) were determined with toluidine blue and van Giesan staining, respectively. The expression of GRP78 and hypoxia-inducible facotr 1α(HIF-1α) was analyzed by the method of immnunohistochemistry. RESULTS: Compared with normal control group at corresponding time point, left ventricular end-diastolic pressure(LVEDP) and ±LV dp/dt_max in 8-week group were significantly decreased (P<0.05). The levels of TNF-α, MDA and CVF, the protein expression of GRP78 and HIF-1α in the myocardial tissues were significantly increased in every model group (P<0.05), and the number of myocardial cells was gradually decreased (P<0.05). Elevated levels of endotoxin in plasma were positively correlated with the levels of alanine aminotransferase (ALT),homocysteine (Hcy) and TNF-α in plasma, the levels of TNF-α, MDA and CVF, and protein levels of GRP78 and HIF-1α in the myocardial tissues (P<0.05). Elevated protein expression of GRP78 in the myocardial tissues was positively correlated with the levels of ALT, Hcy in plasma and MDA, CVF, HIF-1α protein in the myocardial tissues (P<0.05). CONCLUSION: Intestinal endotoxemia induced by liver cirrhosis may directly or indirectly lead to endoplasmic reticulum stress and overexpression of GRP78. GRP78 may be a key molecule in the pathogenesis of myocardial remodeling and functional alteration induced by liver cirrhosis.     

TMT-tagged quantitative proteomics analysis of differentially expressed proteins and enrichment of signaling pathways in nasopharyngeal carcinoma cells with or without SATB1 over-expression

AIM: To analyze the effects of special AT-rich sequence binding protein 1 (SATB1) expression on the protein expression profiles in human nasopharyngeal carcinoma (NPC) cells, and to enrich the differential signaling pathways through bioinformatics analysis. METHODS: SATB1 over-expressing lentivirus and negative control lentivirus were used to infect the CNE1 cells, and then the cell lines were obtained by puromycin stressed method. The total proteins of the 2 cells were extracted, and the differentially expressed proteins were screened by TMT-labeled protein quantification technique and tandem mass spectrometry. The mRNA levels of the differential protein-coding genes were verified by RT-qPCR. GO analysis was used to annotate and enrich the differentially expressed proteins, and the KEGG database was used to enrich and analyze the signaling pathways of differential proteins. RESULTS: SATB1 over-expressing CNE1 cells were established through infected with associated lentivirus. Compared with the control group, 278 differentially expressed proteins were identified in SATB1 over-expressing CNE1 cells, in which 115 were up-regulated and 163 were down-regulated. 10 representative differential protein-coding genes were verified by RT-qPCR, which showed the consistence with the proteomic results. GO analysis indicated differentially expressed proteins were mainly involved in cellular processes, single-organism processes, biological regulation, metabolic processes, protein binding and catalysis. Cell components of differentially expressed proteins mainly existed in cell part, cells and organelles. KEGG analysis showed that differentially expressed proteins were involved in signaling pathways closely related to tumors, includeing MAPK, PI3K-Akt, AMPK, JAK-STAT, p53, PPAR, Hippo and HIF-1 signaling pathways. CONCLUSION: Over-expression of SATB1 significantly alters the protein expression profiles in the NPC cells and affects multiple signaling pathways closely related to tumors. Proteomics also provides a possible macro approach to the screening of molecular mechanisms, therapeutic and prognostic targets for NPC.     

Pharmaceutical inactivation of Erk1/2 does not affect function of FGF21 to regulate glucose and lipid metabolic hemostasis

AIM: To investigate whether inactivation of extracellular signal-regulated kinase 1/2 (Erk1/2) will affect the function of fibroblast growth factor 21 (FGF21) to regulate glucose and lipid metabolism. METHODS: Male db/db mice (8 weeks old) were treated with U0126 (an inhibitor of Erk1/2 kinase) for 1 week, and then treated with recombinant human FGF21 protein and adenovirus-mediated FGF21 (Ad-FGF21). The profile changes of blood glucose and blood lipid were evaluated at 120 min or 4 weeks after FGF21 administration. Meanwhile, the molecular mechanism was explored by in vitro study. RESULTS: Treatment of db/db mice with recombinant human FGF21 protein significantly reduced blood glucose and triglyceride levels at 120 min after FGF21 administration, but these changes were comparable in U0126-treated mice. Furthermore, abnormal glucose and triglyceride levels, and glucose and insulin tolerance were strongly improved in db/db mice as accompanied with decreasing body fat content after 4 weeks of ad-FGF21 administration. Interestingly, treatment with or without U0126 did not influence these effects of FGF21. Mechanically, treatment with Ad-FGF21 significantly upregulated the protein levels of p-Erk1/2 and peroxisome proliferator-activated receptor γ (PPARγ) as well as the expression of adiponectin at mRNA and protein levels in adipose tissues. However, treatment with or without U0126 did not change the profiles. On the other hand, in vitro experiments also indicated that treatment of adipocytes with recombinant human FGF21 protein significantly activated Erk1/2 phosphorylation, and upregulated the expression levels of PPARγ and adiponectin (P<0.05). However, pre-administration of U0126 did not affect the profiles. CONCLUSION: Pharmaceutical inactivation of Erk1/2 by U0216 does not affect the biological function of FGF21 to regulate blood glucose balance and improve abnormal blood lipids in vivo.     

Progress of c-FLIP overexpression in prostatic carcinoma

Cellular FLICE-inhibitory protein (c-FLIP) is a key anti-apoptotic regulator that inhibits death receptor (DR) signal pathway mediated by suppressing procaspase-8 activation. c-FLIP has been found to be elevated in prostatic carcinoma (PCa), and there is a strong correlation between the overexpression of c-FLIP and PCa. In the present paper, we review the links beteween c-FLIP and androgen receptor (AR) signal pathway, which regulate and promote the survival of androgen-independent prostate cancer (AIPC) and describe the possibility of c-FLIP as a therapeutic target of PCa.     

Expression and functional analysis of novel gene AngRem104 associated with glomerular sclerosis

AIM:To study the expression and function of novel gene AngRem104.METHODS:Northern blot was performed to detect the distribution of AngRem104 in human multiple normal tissues as well as the effect of AngⅡ and AT1R antagonist (losartan) on AngRem104 expression. The sense and antisense eukaryotic expression vectors of AngRem104 were constructed and transfected into human mesangial cells. RT-PCR was used to detect the expression of FN when AngRem104 was over-expressed. Primary sequence and motif analysis of AngRem104 protein were performed by on-line ExPasy predictive tools.RESULTS:AngRem104 was predicted to localize at the cellular nucleus. It was widely expressed in human heart, placenta, liver, muscle, kidney and pancreas. Moreover, the up-regulated expression of AngRem104 induced by AngⅡ was inhibited by losartan in a dose-dependent manner.CONCLUSION:AngRem104 is a novel nuclear protein related to the expression of fibronectin and could be up-regulated by AngⅡ in human MC.     

Cross-species comparison of Smad3 expression in hepatocellular carcinoma tissues

AIM： To study the expression of Smad3 and its significance in hepatocellular carcinoma (HCC) in different species including human, rat and tree shrew, and to verify the feasibility of cross-species oncogenomic approach. METHODS：Real-time fluorescent quantitative polymerase chain reaction and Western blotting were applied to detect the expression of Smad3 at mRNA and protein levels in HCC tissues, corresponding HCC adjacent liver tissues and normal liver tissues collected from different species including human, rat and tree shrew. RESULTS：The mRNA expression of Smad3 in HCC tissues of human, rat and tree shrew was lower than those in the corresponding HCC tissues adjacent liver tissues. The mRNA level of Smad3 in HCC tissues of rat was lower than those in its normal liver tissues, while that in HCC adjacent tissues of tree shrew appeared higher than those in the normal liver tissues. No significant difference of Smad3 expression in other tissues was observed. The protein levels of Smad3 in HCC of human and rat were lower than those in the corresponding HCC adjacent liver tissues and the normal liver tissues. However, the protein expression of Smad3 was at a low level in HCC tissues of tree shrew and was lower than that in the HCC adjacent liver tissues and the normal liver tissues, although without statistical difference. The differences of Smad3 expression between HCC adjacent liver tissues and normal liver tissues in all the 3 species were not statistically significant. CONCLUSION：Smad3 is lowly expressed in HCC tissues of different species, suggesting that it might play a pivotal role in hepatocarcinogenesis and be applied as a key molecular target in HCC prevention and treatment.     

Latent membrane protein 1 regulates expression of p27 and RPLP0 in nasopharyngeal epithelial and nasopharyngeal carcinoma tissues

AIM: To observe the protein expression of p27 and ribosomal phosphoprotein large P0(RPLP0) regulated by latent membrane protein 1 (LMP1) in nasopharyngeal epithelial and nasopharyngeal carcinoma tissues. METHODS: The protein levels of p27 and RPLP0 and the relationship with LMP1 were analyzed by Western blotting. The protein expression of LMP1, p27 and RPLP0 was also detected by the method of immunohistochemistry in 30 nasopharyngeal epithelial and 60 nasopharyngeal poorly-differentiated squamous-cell carcinoma tissues. Meanwhile, the significance of clinical pathology was evaluated. RESULTS: The positive rate of LMP1 protein was 73.3% and 90.0% in nasopharyngeal epithelial and nasopharyngeal poorly-differentiated squamous-cell carcinoma tissues, respectively. Compared with the LMP1-negative tissues, the protein levels of RPLP0 were low in the nasopharyngeal epithelial and nasopharyngeal poorly-differentiated squamous-cell carcinoma tissues with LMP1-positive expression, but the levels of RPLP0 protein were overexpressed. The protein expression of RPLP0 and RPLP0 was related to the age of nasopharyngeal carcinoma patients, the protein level of LMP1, the metastasis of lymph nodes and the TNM classification. The positive expression of p27 protein at high level was usually observed in the patients with young age, or had the characteristics of LMP1 (-), non-metastasis of lymph nodes, and in I or II stage of TNM classification. However, the protein expression of RPLP0 was low (P<0.05). CONCLUSION: LMP1 down-regulates p27 and up-regulates RPLP0 in nasopharyngeal epithelial and nasopharyngeal poorly-differentiated squamous-cell carcinoma tissues.