Shelf-life extension of Pacific white shrimp (Litopenaeus vannamei) using sodium alginate/chitosan incorporated with cell-free supernatant of Streptococcus thermophilus FUA 329 during cold storage.

Seafood is highly perishable and has a short shelf-life. This study investigated the effect of chitosan and alginate (CH-SA) coating combined with the cell-free supernatant of Streptococcus thermophilus FUA329 (CFS) as a preservative on the quailty of white shrimp (Litopenaeus vannamei) refrigerated at 4° for 0, 3, 6, 9, 12, 15 days. Freshly shrimps were randomly divided into four groups: the CFS group (400 mL); the CH-SA group (1% chitosan/1% alginate); the CFS-CH-SA group (1% chitosan/1% alginate with 400 mL CFS) are treatment groups, and the control group (400 mL sterile water). The CFS-CH-SA coating effectively suppressed microbial growth total viable count and chemical accumulation (pH, total volatile basic nitrogen, thiobarbituric acid reactive substance) compared with the control. Additionally, the CFS-CH-SA coating improved the texture and sensory characteristics of shrimp during storage. The coated shrimp exhibited significantly reduced water loss (p < 0.05). The combination of CH-SA coating with CFS treatment can extend the shelf life of shrimp. PRACTICAL APPLICATION: Recently, edible films have received more consideration as a promising method to enhance the shelf life of seafood. The presence of Lactic acid bacteria metabolites in edible films reduces spoilage and improves consumer health. Our findings encourage the application of edible coating incorporated with cell-free supernatant of Streptococcus thermophilus FUA 329 to design multifubctional foods and preserve the qualities of shrimp.

Improvement of Urolithin A Yield by In Vitro Cofermentation of Streptococcus thermophilus FUA329 with Human Gut Microbiota from Different Urolithin Metabotypes.

Streptococcus thermophilus FUA329 converts ellagic acid (EA) to urolithin A (Uro-A), which is not autonomously converted by the gut microbiota to produce highly bioavailable and multibiologically active Uro-A in urolithin metabotype 0 (UM-0) populations. We consider that Streptococcus thermophilus FUA329 has the potential to be developed as a probiotic. Therefore, we utilized S. thermophilus FUA329 for in vitro cofermentation with gut microbiota. The results revealed that strain FUA329 increased the production of EA-converted Uro-A during in vitro cofermentation with the human gut microbiota of different urolithin metabotypes (UMs), with a significant increase in the production of Uro-A in the experimental group of UM-0. In addition, changes in the in vitro cofermentation microbial community were determined using high-throughput sequencing. Strain FUA329 modulated the structure and composition of the gut microbiota in different UMs, thereby significantly increasing the abundance of beneficial microbiota in the gut microbiota while decreasing the abundance of harmful microbiota. Of greatest interest was the significant increase in the abundance of Actinobacteria phylum after the cofermentation of strain FUA329 with UM-0 gut microbiota, which might be related to the significant increase in the production of Uro-A.

RNF2 inhibits E-Cadherin transcription to promote hepatocellular carcinoma metastasis via inducing histone mono-ubiquitination.

RNF2 is a RING domain-containing E3 ubiquitin ligase that mediate histone H2A mono-ubiquitination to repress gene transcription, but its expression patterns and molecular function in hepatocellular carcinoma (HCC) remain unclear. Herein, we extracted data from TGCA database and validated RNF2 expression in our own cohort, which revealed that RNF2 was highly expressed in HCC and was associated with malignant characteristics and poor prognosis of HCC. Moreover, RNF2 was demonstrated to promote HCC metastasis via enhancing epithelial-mesenchymal transition (EMT) both in vitro and in vivo. Mechanistically, RNF2 repressed E-Cadherin transcription by increasing the deposition of H2K119ub at the E-Cadherin promoter region. In addition, RNF2-regulated crosstalk between H2AK119ub, H3K27me3 and H3K4me3 synergistically reduced E-Cadherin transcription, which promoted EMT and HCC metastasis. These results indicate that RNF2 played an oncogenic role in HCC progression via inducing EMT, and RNF2 could be a potential therapeutic target for HCC.

Identification of a prognostic cuproptosis-related signature in hepatocellular carcinoma.

BACKGROUND: Cuproptosis is a new type of copper-induced cell death that is characterized by the aggregation of lipoylated tricarboxylic acid (TCA) cycle proteins. However, its role in hepatocellular carcinoma (HCC) remains unclear. The goal of this research is to develop a cuproptosis-related signature predicting the prognosis of HCC. METHODS: The cuproptosis-related genes were defined using Pearson correlation coefficients. LASSO-Cox regression analysis was used to evaluate the prognostic values of cuproptosis-related genes to construct a cuproptosis-related prognostic model. The immune microenvironment analysis was performed by "ssGSEA" to reveal the associated immune cell infiltration patterns with the cuproptosis-related genes signature. The expression levels of one of the prognostic genes PDXK were then verified in HCC samples by Western Blot and immunohistochemistry. The potential roles of target genes in cuproptosis were further explored during in-vitro experiments. RESULTS: A total of 136 cuproptosis-related genes were discovered using Pearson correlation analysis in HCC. A cuproptosis-related signature that included 5 cuproptosis-related genes (PDXK, HPN, SLC25A28, RNFT1, CLEC3B) was established in the TCGA-LIHC training cohort. TCGA validation cohort and another two external validation cohorts confirmed the robustness of the signature's predictive value. Moreover, a nomogram using the risk score was created to best predict the survival of HCC patients. The immune microenvironment analysis revealed distinct immune infiltrations patterns between different risk groups based on the signature model. Furthermore, the upregulation of PDXK was confirmed in HCC tumor tissues in 30 clinical HCC specimens. The knockdown of PDXK reduced the proliferation, migration and invasion of HCC cells. Besides, the expression of PDXK was upregulated after the induction of cuproptosis by elesclomol-CuCL2, which could be suppressed when pretreated with a copper ion chelator. And PDXK deficiency increased the sensitivity of HCC cells to cuproptosis inducer. CONCLUSION: Our study identified a new cuproptosis-related gene signature that could predict the prognosis of HCC patient. Besides, the upregulated PDXK could promote the proliferation and metastasis of HCC. And PDXK deficiency facilities cuproptosis in HCC. Therefore, these fundings highlighted that PDXK might serve as a potential diagnostic and therapeutic target for HCC.

A comprehensive analysis of FOX family in HCC and experimental evidence to support the oncogenic role of FOXH1.

Hepatocellular carcinoma (HCC) remains the second leading cause of cancer related deaths worldwide. Understanding about the molecular biology of HCC and development of targeted therapies are still the main focuses of this type of disease. Here, by connecting the expression levels of FOX proteins with their associated clinical characteristics using TCGA LIHC dataset, we found that 27/40 FOX proteins were highly expressed in HCC tumors compared to normal liver tissues and their expression levels were tightly associated with HCC tumor stage, tumor grade and overall survival. Our experimental results also confirmed that FOXH1 indeed played an oncogenic role in HCC development by promoting cell growth and cell migration/invasion. Mechanistic dissection demonstrated that FOXH1-induced cell growth and cell migration/invasion relied on mTOR signaling because inhibition of mTOR signaling by rapamycin could attenuate FOXH1-mediated phenotypic alterations of HCC cells. The results from orthotopic mouse model also validated that FOXH1 promoted HA22T tumor growth via triggering mTOR activation. Overall, this study not only comprehensively examines the clinical values of FOX proteins in HCC but also provides experimental evidence to support the role of FOXH1 in HCC development, building rationale to develop more effective therapies to treat HCC patients.

Testicular orphan receptor 4 (TR4) promotes papillary thyroid cancer invasion via activating circ-FNLA/miR-149-5p/MMP9 signaling.

The incidence and mortality of papillary thyroid cancer (PTC) have steadily increased. Although conventional therapies are very effective toward differentiated PTC patients, very limited therapeutic options are applicable to those patients with distant metastases. Therefore, better understanding of the molecular biology of metastatic PTC helps identify novel targets and facilitates the development of new therapies. In this study, we first found that testicular orphan receptor 4 (TR4) was significantly increased in PTC tumors spreading to lymph nodes compared to the paired primary tumors. Experimental evidence suggested that TR4 drove PTC progression via promoting its cell invasion and cell migration. Mechanistically, TR4 transcriptionally regulated the expression level of circ-filamin A (FLNA), which competed with matrix metalloproteinase 9 (MMP9) for microRNA (miR)-149-5p binding and led to an increased protein level of MMP9. Interruption assays with various gene manipulations verified that the TR4/circ-FLNA/miR-149-5p/MMP9 signaling axis played a central role in cell invasion and cell migration of PTC cells. Moreover, a xenografted mouse model also confirmed that the TR4/circ-FLNA signal promoted PTC tumor growth. Overall, our study pinpoints the oncogenic role of TR4 in PTC development, and the targeting of TR4/circ-FLNA/miR-149-5p/MMP9 signaling may be an alternative option for metastatic PTC patients.

Androgen receptor (AR) decreases HCC cells migration and invasion via miR-325/ACP5 signaling.

Hepatocellular carcinoma (HCC) is the most 5th commonly diagnosed and 2nd most lethal tumor in the world. The obvious gender advantage of HCC indicates that androgen receptor (AR) may play an important role in the tumor occurrence, develop and metastasis of HCC. Here we found that decreased AR could alter miR-325 to increase ACP5 expression in HCC cells, to increase HCC cells migration and invasion capacities. Mechanism dissection revealed that AR could regulate miR-325 expression through transcriptional regulation and miR-325 might directly target the 3'UTR of ACP5-mRNA to suppress its translation. The in vivo orthotopic xenografts mouse model with oemiR-325 also validated in vitro data. Together, these findings suggest that AR may decrease HCC progression through miR-325/ACP5 signaling and targeting the AR/miR-325/ACP5 signaling may help in the development of the novel therapies to better suppress the HCC progression.

Elevated CELSR3 expression is associated with hepatocarcinogenesis and poor prognosis.

Cadherin EGF LAG seven-pass G-type receptor 3 (CELSR3) has been reported to exhibit a cancer-specific pattern. The present study aimed to investigate the clinical value and functional role of CELSR3 in hepatocellular carcinoma (HCC), and determine the underlying molecular mechanism in patients with HCC. CELSR3 expression in tumor and paracancerous HCC tissues was obtained from The Cancer Genome Atlas. Differential expression analysis was performed using the edgeR package. Pearson correlation analysis was used to analyze the correlation between methylation and mRNA levels of CELSR3. Pathways affected by aberrant CELSR3 expression were identified through Gene Set Enrichment Analysis. The results demonstrated that CELSR3 was highly expressed in the early stage of cancer and was present throughout the entire cancer process, which suggested that CELSR3 may serve a key role in the carcinogenesis of HCC. In addition, upregulation of CELSR3 was associated with its methylation level; high CELSR3 expression indicated a shorter overall survival time. Multiple candidate genes were screened by integrating differentially expressed (DE) mRNAs and target genes of DE microRNAs (miRs). Subsequent pathway enrichment analysis demonstrated that the upregulated genes were predominantly enriched in the 'Neuroactive ligand-receptor interaction' and 'Cell cycle' pathways, whereas the downregulated genes were primarily enriched in 'Cytokine-cytokine receptor interaction' and 'Metabolic pathways'. CELSR3 and its connected nodes and edges were initially removed from the miRNA-mRNA regulatory network in order to prevent bias and compared with the network containing CELSR3 alone. The frequently dysregulated miRNAs were identified as miR-181 family members, and the results suggested that miR-181 and the Wnt/ß-catenin signaling pathway influenced CELSR3 expression.

SEMA3F Promotes Liver Hepatocellular Carcinoma Metastasis by Activating Focal Adhesion Pathway.

Previous studies have shown that semaphorin-3F (SEMA3F) functions as a tumor suppressor in several tumor types. However, the role of SEMA3F in the metastasis and prognosis of liver hepatocellular carcinoma (LIHC) remains unknown. In this study, by performing bioinformatics analysis on the transcriptome profiles from The Cancer Genome Atlas (TCGA), we demonstrated that SEMA3F was significantly upregulated in LIHC tissues, compared with normal controls. Moreover, the expression value of SEMA3F was positively correlated with patients' pathological stages and tumor metastasis, predicting a poor overall survival. Besides, SEMA3F expression level was negatively correlated with its methylation level, but positively correlated with its gene copy number. Differential expression analysis of LIHC samples with high or low SEMA3F expression values suggested that 983 genes were differentially expressed, among which 723 genes were upregulated and 260 genes were downregulated. Furthermore, enrichment analysis of differentially expressed genes revealed that SEMA3F was involved in the activation of focal adhesion pathway, which induced tumor metastasis. Taken together, our results suggested that the oncogenic function of SEMA3F promoted hepatocellular carcinoma metastasis by activating focal adhesion pathway.

MiR-425-5p promotes invasion and metastasis of hepatocellular carcinoma cells through SCAI-mediated dysregulation of multiple signaling pathways.

MicroRNAs (miRNAs) play critical roles in hepatocellular carcinoma (HCC) progression and are key determinants of prognosis. In this study, we found that miR-425-5p was elevated in HCC and correlated with poor prognostic clinicopathological features and low post-operative long-term survival. Multivariate survival analysis indicated that miR-425-5p expression was an independent risk factor for overall and disease-free survival. Interestingly, miR-425-5p promoted invasion and metastasis by HCC cells, but not HCC cell proliferation or apoptosis in vitro. SCAI and PTEN were determined to be downstream targets of miR-425-5p. miR-425-5p-mediated effects were inhibited by ectopic expression of SCAI, and PTEN exhibited a smaller inhibitory effect. SCAI also suppressed PTEN expression. In addition, miR-425-5p promoted epithelial-to-mesenchymal transition (EMT), which was antagonized by SCAI. miR-425-5p also promoted HCC cell invasion and metastasis via SCAI-mediated dysregulation of integrin ß1-Fak/Src-RhoA/CDC42, PTEN-AKT, and TIMP2-MMP2/MMP9 signaling. Finally, miR-425-5p promoted metastasis in a xenograft mouse model of HCC. These results indicate that miR-425-5p facilitates EMT and extracellular matrix degradation and promotes HCC metastasis through SCAI-mediated dysregulation of multiple signaling pathways. MiR-425-5p is therefore a potential prognostic biomarker and novel therapeutic target in HCC.

The lncRNA XIST interacts with miR-140/miR-124/iASPP axis to promote pancreatic carcinoma growth.

Long non-coding RNA (lncRNA) X-inactive specific transcript (XIST) is involved in the development and progression of many tumors. In this study, XIST was specifically upregulated in pancreatic carcinoma tissues and cell lines; a higher XIST expression was correlated to poorer clinicopathologic features. After XIST knockdown, the proliferation of PC cell lines was suppressed and cell cycle stagnated in G1 phase; XIST knockdown also reduced the protein levels of inhibitor of apoptosis-stimulating protein of p53 (iASPP) and Cyclin-dependent kinase 1 (CDK1), increased the protein level of P21, a potent CDK inhibitor. In PC cell lines, XIST and miR-140/miR-124, two tumor-associated miRNAs, could inversely regulate each other, respectively; miR-140/miR-124 could bind to XIST and the 3'UTR of PPP1R13L, respectively. XIST and miR-140/miR-124 exerted opposite effects on iASPP, CDK1, P21 and P27 proteins; whereas the effects of LV-sh-XIST on the indicated protein levels could be partially reversed by miR-140 and/or miR-124 inhibitor. In PC tissues, miR-140 and miR-124 expression was down-regulated, iASPP and CDK1 mRNA expression was up-regulated. XIST positively correlated with iASPP and CDK1, inversely correlated with miR-140 and miR-124, respectively. Taken together, our data indicated that XIST might be an oncogenic lncRNA that promoted proliferation of PC cell line through inhibiting miR-140/miR-124 expression and promoting cell cycle-related factor expression, and could be regarded as a therapeutic target in human pancreatic carcinoma.

MicroRNA-140 regulates cell growth and invasion in pancreatic duct adenocarcinoma by targeting iASPP.

MicroRNAs are ∼22 nucleotide RNAs processed from RNA hairpin structures that play important roles in regulating protein expression level via binding to mRNA, either suppressing its translation or speeding up its degradation. In humans, they regulate most protein-coding genes, including genes important in cancer and other diseases. In this study, the expression of microRNA-140 (miR-140) was demonstrated to be significantly suppressed in pancreatic duct adenocarcinoma specimens and cell lines, compared with their adjacent normal tissues. With the help of bioinformatics analysis, inhibitor of apoptosis-stimulating protein of p53 (iASPP) was identified to be a direct target of miR-140, and luciferase reporter experiment confirmed this discovery. Overexpression of miR-140 decreases the protein expressions of iASPP, ΔNp63, MMP2, and MMP9. Growth and invasion of PANC-1 cells were attenuated by overexpression of miR-140 in vitro. The suppressive effect of miR-140 on PANC-1 cell line could be partly balanced out by manual overexpression of iASPP. Above all, these findings provided insights into the functional mechanism of miR-140, suggested that the miR-140/iASPP axis may interfere with the proliferative and invasive property of pancreatic duct adenocarcinoma cells, and indicated that miR-140 could be a potential therapeutic target for pancreatic duct adenocarcinoma.

DNA cytosine and methylcytosine deamination by APOBEC3B: enhancing methylcytosine deamination by engineering APOBEC3B.

APOBEC (apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like) is a family of enzymes that deaminates cytosine (C) to uracil (U) on nucleic acid. APOBEC3B (A3B) functions in innate immunity against intrinsic and invading retroelements and viruses. A3B can also induce genomic DNA mutations to cause cancer. A3B contains two cytosine deaminase domains (CD1, CD2), and there are conflicting reports about whether both domains are active. Here we demonstrate that only CD2 of A3B (A3BCD2) has C deamination activity. We also reveal that both A3B and A3BCD2 can deaminate methylcytosine (mC). Guided by structural and functional analysis, we successfully engineered A3BCD2 to gain over two orders of magnitude higher activity for mC deamination. Important determinants that contribute to the activity and selectivity for mC deamination have been identified, which reveals that multiple elements, rather than single ones, contribute to the mC deamination activity and selectivity in A3BCD2 and possibly other APOBECs.

WD40 repeat-containing 62 overexpression as a novel indicator of poor prognosis for human gastric cancer.

AIM: WD40 repeat-containing 62 (WDR62) is a centrosome-associated gene involved in cell cycling and proliferation. However, the role of WDR62 in human malignancies remains unknown. The present study aimed to identify the role, if any, of WDR62 in the pathogenesis of human gastric cancer (GC). METHODS: WDR62 expression in 372 cases of human GC and eight GC cell lines was determined using quantitative reverse transcription-polymerase chain reaction (qRT-PCR), immunohistochemistry and Western blotting. Correlations between WDR62 expression and clinicopathological characteristics, as well as GC prognosis were determined. WDR62 regulation of GC cell proliferation, invasion, migration and cell cycle distribution were studied both in vitro and in vivo. RESULTS: WDR62 expression was significantly increased in GC tissues and cell lines and was associated with poor differentiation and prognosis of GC. WDR62 expression was elevated in GC multidrug resistant cells. Suppressing WDR62 significantly decreased cell proliferation and induced G2/M phase arrest of GC cells. Consistently, WDR62 knockdown inhibited gastric carcinogenesis in nude mice. Regulation of Akt/p38-mitogen-activated protein kinase (MAPK)/multidrug resistance gene 1 (MDR1) expression and activation by WDR62 contributed to the chemoresistance of GC cells. WDR62 overexpresses in GC and the suppression of WDR62 inhibits GC cell growth by inducing G2/M cell cycle arrest. CONCLUSION: WDR62 may be a novel prognostic marker and a potential chemotherapy target for GC.

Overexpression of MMSET is correlation with poor prognosis in hepatocellular carcinoma.

The multiple myeloma SET domain (MMSET) involved in the t(4;14)(p16;q32) chromosomal translocation encodes a histone lysine methyltransferase. High expression of MMSET is common translocation in multiple myeloma (MM) and is associated with the worst prognosis. Recent studies have shown that overexpression of MMSET is significant in other tumor types compared to their normal tissues. However, little is known about its role in hepatocellular carcinoma (HCC). In these study we investigate the expression of MMSET in HCC and to make correlations with clinicopathologic features. Twenty-eight pairs of HCC and adjacent non-tumor tissues, and eight normal liver tissues were collected for MMSET detection by western blotting and real time-PCR analysis. Immunohistochemistry was used to determine the expression of MMSET in HCC and adjacent non-tumor tissues from 103 patients. Overexpression of MMSET was significantly associated with Edmondson stage, vascular invasion. Moreover, Kaplan-Meier curves showed that MMSET upregulated was associated with shorter overall survival and disease-free survival in HCC patient. In conclusion, our study demonstrates for the first time that overexpression of MMSET is an independent prognostic factor and is correlated with poor survival in HCC patients.

[Construction and application of a noval gene-target therapy system in hepatocellular carcinoma].

This paper is aimed to explore the efficiency of a noval gene-target therapy system in hepatocellular carcinoma (HCC) treatment in vitro, by using survivin gene as the target. A new fusion promoter (AV) driving pcDNA3. 1 (-)AV plasmid, which contained the cytomegalovirus (CMV) enhancer and alpha-fetoprotein (AFP) promoter, was constructed by molecular biologic method. The eukaryotic expression plasmid pcDNA3. 1(-)AVGFP and pcDNA3. 1 (-)AVsiRNA-survivin were constructed by cloning and inserting the green fluorescent protein (GFP) and siRNA-survivin sequence into pcDNA3. 1(-) AV plasmid separately. Then these two plasmids were transfected into HepG2, SMMC-7721 and Hela cells by using nanoparticles of calcium phosphate. The transfection efficiencies were detected by GFP. Reversed transcript polymerase chain reaction (RT-PCR) and western-blot were used to evaluate the knockdown efficiency of siRNA-survivin. The growth curves and cell death of HepG2 cells transfected with or without pcDNA3. 1(-)AVsiRNA-survivin were detected by MTT assay and flow cytometry assay, respectively. The results showed that after transfected with pcDNA3. 1(-)AVGFP vector, only HepG2 cells displayed strong GFP signaling, whereas, no GFP was found in Hela cells, suggesting that AV promoter can specifically drive downstream of gene expression in HCC cells. Furthermore, the mRNA and protein expression levels of survivin in HepG2 cells, but not in Hela and SMMC-7721 cells, were significantly silenced after pcDNA3. 1(-)AVsiRNA-survivin transfection. Finally, compared with the control cells, HepG2 cells, which were transfected with pcDNA3. 1(-) AVsiRNA-survivin plasmid, presented 68.8% cell death, including 38.68% apoptosis and 30.12% necrosis, and significant cell growth inhibition (P < 0.05). These findings indicated that this noval gene-target therapy system could specifically target HCC cells with high efficiency, providing a new gene therapy strategy for HCC.

Downregulation of MAGI1 associates with poor prognosis of hepatocellular carcinoma.

OBJECTIVE: MAGI1 (membrane-associated guanylate kinase, WW and PDZ domain containing 1) plays an important role in stabilization of adherens junctions and suppression of invasiveness and metastasis. However, its expression and clinical revelance in hepatocellular carcinoma (HCC) are still unknown. So, this study was performed to detect the expression and clinical significance of MAGI1 in human HCC. METHODS: The mRNA and protein expression levels were examined by using reverse transcription-PCR (RT-PCR) and Western blotting in 31 paired HCC and paracarcinomatous liver tissues (PCLT). Furthermore, samples of 104 HCC patients were determined immunohistochemically for MAGI1 expression and the correlation of MAGI1 levels with prognosis was analyzed. RESULTS: The expression levels of MAGI1 were significantly downregulated in HCCs than those in PCLTs (p < .01). Importantly, the decreased expression of MAGI1 correlated with multiple tumor nodules (p < .05), absence of capsular formation (p < .05), worse Edmondson-Steiner grade (p < .05), vein invasion (p < .01), shortened median overall survival time (25 versus 42 months; p = .013), and disease-free survival time (20 versus 40 months; p = .018) of HCC. Multivariable Cox regression analysis revealed that MAGI1 expression level was an independent factor for prognosis (p = .038). CONCLUSIONS: MAGI1 expression is decreased in HCC, which correlates with poor prognosis, suggesting MAGI1 as a novel prognostic marker for HCC.

MAGI1 inhibits cancer cell migration and invasion of hepatocellular carcinoma via regulating PTEN.

OBJECTIVE: To explore the biological function and molecular mechanism of membrane associated guanylate kinase, WW and PDZ domain containing 1 (MAGI1) in hepatocellular carcinoma. METHODS: HepG2(MAGI1) stable cell line was constructed by transfecting HepG2 cells with pcDNA3.1-MAGI1 plasmid. Wound healing and invasion assay were performed to compare the migration and invasion ability of HepG2(MAGI1) and HepG2 cells. Furthermore, the expression of MAGI1 and phosphatase and tensin homolog deleted on chromosome ten (PTEN) was also examined by Western blot and the relationship was analyzed. RESULTS: The wound healing assay showed that the closure of HepG2(MAGI1) cells was significantly slower than that of HepG2 cells [(90 ± 10)% vs. (50 ± 15)%, P<0.05], and the invasion assay showed that the number of HepG2(MAGI1) cells that passed through the matrigel was fewer than HepG2 cells (68 ± 18 vs. 150 ± 30, P<0.05). The protein expression level of PTEN was significantly elevated in HepG2(MAGI1) cells compared with HepG2 cells (1.40 ± 0.32 vs. 0.28 ± 0.15, P<0.05). MAGI1 and PTEN protein expression levels were positively correlated (r=0.913, P<0.01). CONCLUSION: MAGI1 may inhibit the cancer cell migration and invasion of hepatocellular carcinoma via regulating PTEN.

[RNA interference of HDAC7 expression in hepatocellular carcinoma].

OBJECTIVE: To investigate the role of histone deacetylase 7 (HDAC7) in the occurrence and development of hepatocellular carcinoma. METHODS: HepG2 cells and human microvascular endothelial cells (HMEC-1) were divided into 3 groups after transfection pSUPER-HDAC7 retroviral interference plasmid: a pSUPER(HDAC7RNAi+) group, a pSUPER(HDAC7RNAi-) group, and a pSUPER group as the control group. The expression of HDAC7, p21, cyclin E, matrix metalloproteinases 10 (MMP10), and hypoxiainducible factor-1alpha (HIF-1alpha) were detected by Western blot. The expression of HDAC7 in cell lines was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry, the nude mice modle, and vascular endothelial cells 2-dimensional tubulogenesis in vitro and in vivo. RESULTS: Compared with the control group and the pSUPER(HDAC7RNAi-) group, the expression of HDAC7 was downregulated, the rate of cell growth inhibition and apoptosis in the pSUPER(HDAC7RNAi+) group increased more significantly; the expression of p21 and HIF-1alpha was increased significantly, while the expression of cyclin E and MMP10 in the pSUPER(HDAC7RNAi+) group was downregulated (P<0.05). CONCLUSION: The expression of HDAC7 protein plays an important role in the apoptosis and vascular tubulogenesis of hepatocellular carcinoma by the upregulation of p21 and HIF-1alpha and the downregulation of cyclin E and MMP10.

Prognostic significance of neuroepithelial transforming protein 1 in hepatocellular carcinoma.

OBJECTIVE: Recent works have demonstrated that neuroepithelial transforming protein 1 (NET-1) gene expression is associated with proliferation, metastasis, and clinical stages of human hepatocellular carcinoma (HCC). To investigate its prognostic significance in HCC, which currently is unknown, the authors examined the correlation between NET-1 expression and prognosis in patients with HCC. METHODS: Immunohistochemical staining was used to determine NET-1 expression level in 368 paired HCC and normal liver tissue (NLT) specimens. Kaplan-Meier survival and Cox regression analyses were performed to evaluate the prognosis of HCC. RESULTS: Among 368 specimens of HCC, the positive rate of NET-1 protein expression in HCC tissues was 86.7% (319/368), and the increased NET-1 expression was correlated significantly with high Edmondson-Steiner grade (p = .02) and TNM stage (p = .01). The expression level of NET-1 in HCC tissues was associated with intrahepatic metastasis (p = .008) and portal vein infiltration (p = .007). HCC patients with the moderate-strong NET-1 positive expression had either poorer disease-free survival or poorer overall survival than patients with negative-low positive NET-1 expression (p = .001 and .002, respectively). A multivariate Cox regression analysis revealed that NET-1 protein expression (relative risk 5.8; p = .01) was an independent prognostic factor for patients with HCC. CONCLUSION: Taken together, our study suggests for the first time that NET-1 status may be a new predictor of survival for HCC patients and provides the rationale for developing a novel therapy of targeting NET-1 against this fatal malignancy.