The clinical and prognostic significance of LGR5 in GC: A meta-analysis of IHC assay and bioinformatics analysis.

Recently, leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5) is a newly identified cancer stem cell marker and Wnt target gene. However, the role of LGR5 in gastric cancer (GC) remains uncertain. This study was performed to investigate the effect of LGR5 expression in GC. The eligible studies were searched via electronic databases. The odds ratios (ORs) with 95% confidence intervals (CIs) or hazard ratios (HRs) with 95% CIs were applied to estimate the effect of LGR5. Further bioinformatics validation data were used to confirm our results. Eleven studies consisting of 2,646 GC patients were identified. LGR5 expression was not associated with age, gender, tumor stage, T stage, tumor size, lymphatic invasion, lymph node metastasis, and distal metastasis. LGR5 expression was related to tumor type (intestinal vs. diffuse: OR=2.25, p=0.032). LGR5 expression was negatively correlated with tumor grade (grade 3-4 vs. grade 1-2: OR = 0.40, p=0.033). Further TCGA validation data also showed similar findings, and LGR5 expression was also found to have a negative association with tumor grade. LGR5 expression was associated with worse overall survival (OS) using multivariate Cox analysis (HR=2.54, p=0.009). Further bioinformatics data showed that LGR5 expression was still correlated with shorter OS in 876 GCs. LGR5 expression was negatively correlated with tumor grade and its expression was higher in intestinal-type than in diffuse-type. Moreover, LGR5 may be a potential prognostic factor for survival prediction in GC.

MicroRNA-410 functions as a tumor suppressor by targeting angiotensin II type 1 receptor in pancreatic cancer.

MicroRNAs (miRNAs) act as key regulators of gene expression in diverse biological processes and are intimately involved in tumorigenesis. However, the underlying molecular mechanisms of miR-410 in pancreatic cancer remain poorly understood. In this study, we found that miR-410 overexpression suppressed pancreatic cancer cell growth in vitro and in vivo as well as cell invasion and migration. miR-410 also resulted in G1/S cell-cycle arrest. We then showed that angiotensin II type 1 receptor (AGTR1) was a direct target of miR-410, with miR-410 suppressing AGTR1 expression levels. In contrast, inhibition of miR-410 increased the expression of AGTR1. Silencing of AGTR1 inhibited cell growth and invasion, similar to miR-410 overexpression. In addition, we found that the induction of vascular endothelial growth factor and the activation of the ERK signaling pathway by angiotensin II were blocked by miR-410, similar to the angiotensin II inhibitor losartan. miR-410 overexpression inhibited angiogenesis in mice through the repression of CD31 expression. ERK pathway knockdown suppressed pancreatic cancer cell proliferation, invasion, and angiogenesis. Finally, we found that miR-410 was downregulated in pancreatic cancer tissues compared to adjacent nontumor tissues, whereas AGTR1 was upregulated in pancreatic cancer tissues. Pearson correlation analysis showed that miR-410 and AGTR1 were inversely expressed. In conclusion, our data indicate that miR-410 suppresses pancreatic cancer growth, cell invasion, migration, and angiogenesis via the downregulation of AGTR1, acting as a tumor-suppressive miRNA. In addition, our results suggest that miR-410 is a potential diagnostic biomarker and therapeutic target for patients with pancreatic cancer.

3-O-Methyl-D-Glucose Blunts Cold Ischemia Damage in Kidney via Inhibiting Ferroptosis.

BACKGROUND: The glucose derivative 3-O-methyl-D-glucose (OMG) is used as a cryoprotectant in freezing cells. However, its protective role and the related mechanism in static cold storage (CS) of organs are unknown. The present study aimed to investigate the effect of OMG on cod ischemia damage in cold preservation of donor kidney. METHODS: Pretreatment of OMG on kidney was performed in an isolated renal cold storage model in rats. LDH activity in renal efflux was used to evaluate the cellular damage. Indicators including iron levels, mitochondrial damage, MDA level, and cellular apoptosis were measured. Kidney quality was assessed via a kidney transplantation (KTx) model in rats. The grafted animals were followed up for 7 days. Ischemia reperfusion (I/R) injury and inflammatory response were assessed by biochemical and histological analyses. RESULTS: OMG pretreatment alleviated prolonged CS-induced renal damage as evidenced by reduced LDH activities and tubular apoptosis. Kidney with pCS has significantly increased iron, MDA, and TUNEL+ cells, implying the increased ferroptosis, which has been partly inhibited by OMG. OMG pretreatment has improved the renal function (p <0.05) and prolonged the 7-day survival of the grafting recipients after KTx, as compared to the control group. OMG has significantly decreased inflammation and tubular damage after KTx, as evidenced by CD3-positive cells and TUNEL-positive cells. CONCLUSION: Our study demonstrated that OMG protected kidney against the prolonged cold ischemia-caused injuries through inhibiting ferroptosis. Our results suggested that OMG might have potential clinical application in cold preservation of donor kidney.

MicroRNA-182 promotes cell growth, invasion, and chemoresistance by targeting programmed cell death 4 (PDCD4) in human ovarian carcinomas.

As an important tumor suppressor, programmed cell death 4 (PDCD4) influences transcription and translation of multiple genes, and modulates different signal transduction pathways. However, the upstream regulation of this gene is largely unknown. In this study, we found that microRNA-182 (miRNA-182, miR-182) was upregulated, whereas PDCD4 was downregulated in ovarian cancer tissues and cell lines. Blocking or increase of miR-182 in ovarian cancer cell lines led to an opposite alteration of endogenous PDCD4 protein level. Using fluorescent reporter assay, we confirmed the direct and negative regulation of PDCD4 by miR-182, which was dependent on the predicted miR-182 binding site within PDCD4 3' untranslated region (3' UTR). MTT and colony formation assays suggested that miR-182 blockage suppressed, whereas miR-182 mimics enhanced viability and colony formation of ovarian cancer cells. These effects may partly be attributed to the cell cycle promotion activity of miR-182. miR-182 also contributed to migration and invasion activities of ovarian cancer cells. Furthermore, miR-182 reduced the chemosensitivity of ovarian cancer cells to CDDP and Taxol, possibly by its anti-apoptosis activity. Importantly, all the alterations of the above cellular phenotypes by blocking or enhancing of miR-182 could be alleviated by subsequent suppression or ectopic expression of its target PDCD4, respectively. We conclude that in ovarian cancer cells, miR-182 acts as an oncogenic miRNA by directly and negatively regulating PDCD4.

[Effect of 5-aza-2'-deoxycytidine on growth and methylation of RUNX3 gene in human pancreatic cancer cell line MiaPaca2].

OBJECTIVE: To investigate the effect of demethylating agent 5-aza-2'-deoxycytidine (5-Aza-CdR) on the growth of human pancreatic cancer cell line MiaPaca2 and the expression and methylation of tumor suppressor gene RUNX3. METHODS: Human pancreatic cancer cell line MiaPaca2 cells were treated with different concentrations of 5-Aza-CdR. Morphological changes of MiaPaca2 cells were observed by light microscopy. The activity of cell proliferation was analyzed by MTT assay. The changes of RUNX3 mRNA expression were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Changes of RUNX3 gene methylation was detected by methylation-specific polymerase chain reaction. RESULTS: MiaPaca2 cells were treated with 2.5, 5, 10 and 20 µmo1/L 5-Aza-CdR, respectively. The inhibition rates of MiaPaca2 cells treated for 24 h were (9.17 ± 2.15)%, (10.75 ± 2.04)%, (12.57 ± 1.64)% and (18.70 ± 1.51)%, respectively. The inhibition rates were (14.94 ± 1.68)%, (18.60 ± 1.57)%, (22.84 ± 1.58)% and (33.24 ± 1.53)%, respectively, after 48 h treatment; (21.46 ± 1.60)%, (28.62 ± 1.72)%, (35.14 ± 1.64)% and (45.06 ± 1.47)%, respectively, after 72 h treatment; and (26.35 ± 1.71)%, (34.48 ± 1.69)%, (40.05 ± 1.60)% and (49.99 ± 1.61)%, respectively, after 96 h treatment. The differences between inhibition rates of each experimental and control groups (0.00 ± 0.00)% were statistically significant (P < 0.05). At the same time, the inhibition rates of different concentration groups showed significant differences (P < 0.05). At 48 h, 72 h and 96 h, the inhibition rates of each pair concentration groups showed significant differences (P < 0.05). 5-Aza- CdR inhibited the growth of MiaPaca2 cells, and the higher the concentration, the stronger the inhibition after 24 h. 5-Aza-CdR also reversed the methylation status of RUNX3 gene, and restored the expression of RUNX3 mRNA with a dose-effect relationship. CONCLUSIONS: The methylation of RUNX3 gene is significantly related with the occurrence and development of pancreatic cancer, and abnormal methylation of RUNX3 gene may contribute to the loss of RUNX3 mRNA expression. 5-Aza-CdR may effectively cause reversion of RUNX3 methylation, and treatment with 5-Aza-CdR can reactivate the gene expression and inhibit the cell growth. This may provide a new way for diagnosis and treatment of pancreatic cancer.

NKD2 is correlated with the occurrence, progression and prognosis of thyroid carcinoma.

BACKGROUND: Thyroid carcinoma (THCA) is the most prevalent type of tumor in endocrine system. NKD2 has been increasingly evidenced to play crucial roles in many cancers, except for THCA. We herein aimed to explore the potential role of NKD2 in THCA. METHODS: Totally 502 THCA patient data were downloaded from TCGA (The Cancer Genome Atlas) database. Overall survival was estimated by Kaplan-Meier method. Gene set enrichment analysis was conducted to obtain significant functional pathways. Wilcoxon rank sum test was used to determine the NKD2 expression differences among various groups. The NKD2 expression was validated in cell lines and tissue microarray. RESULTS: Significantly higher NKD2 expression was observed in THCA samples compared with adjacent samples, which were successfully verified in cell lines and tissue microarray. Moreover, NKD2 expression gradually elevated along with the increase of TNM Stage, and NKD2 expression was significantly higher in elder THCA patients compared with young patients. NKD2 highly expressed THCA patients had worse prognosis compared with NKD2 low-expressed patients. Furthermore, 53 pathways were significantly activated in the high NKD2 expression patients compared with low NKD2 expression THCA patients. CONCLUSIONS: In summary, high NKD2 expression was probably related to the progression and poor prognosis of THCA. NKD2 is a promising prognostic biomarker and pathogenic target of THCA.

FGF Exhibits an Important Biological Role on Regulating Cell Proliferation of Breast Cancer When it Transports Into The Cell Nuclei.

The endocrine system is closely related to the development of the breast cancer. Many studies have shown that FGF1 (Fibroblast growth factor-1) is involved the occurrence and development of the breast cancer. But up to now, the cellular behavior and characteristics of FGF1 in breast cancer have not been fully revealed. In the current study, breast cancer cell was used as an in vitro cell model to investigate FGF's cell property. The results showed that FGF1 internalized into cells in a time-dependent manner. Further study indicated that both clathrin-mediated and caveolin-mediated endocytic pathway are involved in the internalization of FGF/FGFR (Fibroblast growth factor receptor), and both clathrin-mediated endocytosis and caveolin-mediated endocytosis are involved in the process of FGF1's nuclear localization. Further study showed that Rab5 also plays an important role in the process of nuclear localization of FGF-1. In addition, we found that FGF1 and FGFR transported to the cell nuclei of breast cancer. Further experimental results indicated that the nuclear-localized FGF1 and/or FGFR is closely associated to cell proliferation of breast cancer cell. Taken together, the current work lays the foundation for exploring the relationship between nuclear-localized FGF1/FGFR and the occurrence and development of breast cancer.

Effects of prolonged cold ischemia on the DCD kidney function and Inflammasome expression in rat kidney transplants.

BACKGROUND: Acute kidney injury (AKI) is the main reason for the bad outcome of the donation of circulatory death (DCD) kidney after transplantation. Prolonged cold storage (CS) is a risk factor for the occurrence of the delayed graft function in DCD kidney. The protein NLR-domain containing receptor 3 (NLRP3) plays a crucial role in renal ischemia reperfusion injury by triggering inflammasome formation. Herein, we investigated whether the NLRP3 signal participate in the CS-induced damage of DCD kidney in rat kidney transplantation models. MATERIALS AND METHODS: DCD kidney and living donor (LD) kidney of SD rats were preserved in UW solution at 4 °C for 2 h or 18 h, and then transplanted into syngeneic recipient. Thus, the animals were randomly divided into 4 groups: 2-h LD group, 2-h DCD group, 18-h LD group and 18-h DCD group. The renal function and pathological changes were determined. The expressions of NLRP3 and inflammatory factor IL-1ß were assessed. The concentration of ferrous iron (Fe2+) was analyzed both in kidneys and in the preservation solution. The renal morphological changes were examined by hematoxylin eosin staining. RESULTS: Our results showed that the levels of Cr and BUN were higher in 18-h LD group as compared to the 2-h LD group, which were remarkably increased in 18-h DCD group. The expression levels of NLRP3 and IL-1ß were increased by 18-h CS compared to 2-h CS in both LD kidney and DCD kidney. In addition, the Fe2+ concentration has significantly increased in 18-h LD group than that in 2-h LD group, and the elevation of Fe2+ was more remarkable in DCD kidneys. CONCLUSION: In conclusion, our study demonstrated that prolonged hypothermic storage of DCD kidney deteriorated the graft function via the increased Fe2+ concentration, which was associated with the upregulation of NLRP3 expression.

Detection of MAGE-1, MAGE-3 and AFP mRNA as multimarker by real-time quantitative PCR assay: a possible predictor of hematogenous micrometastasis of hepatocellular carcinoma.

BACKGROUND/AIMS: The aim of this study was to explore the relationship between MAGE-1, MAGE-3 and AFP mRNA in the peripheral blood of patients with hepatocellular carcinoma and micrometastasis in circulation, real-time quantitative-PCR (real-time Q-PCR) assay was applied to detect the expression of the multimarker. METHODOLOGY: Peripheral blood samples were obtained from 86 patients with hepatocellular carcinoma (HCC) and real-time Q-PCR technique was used to detect the MAGE-1, MAGE-3, and AFP mRNA in the blood. RESULTS: In 86 tumor specimens, the positivity for MAGE-1, MAGE-3, and AFP genes was 34.9% (30/86), 60.5% (52/86) and 69.8%(60/86) respectively, and all specimens expressed at least one marker. MAGE-1, MAGE-3, and AFP transcripts were detected in 12 (14.0%),18 (20.1%) and 29 (33.7%) of 86 blood specimens from hepatocellular carcinoma patients, respectively, while 45 specimens (52.3%) were positive for at least one marker. In addition, MAGE-1, MAGE-3 and AFP gene transcripts were not detected in any peripheral blood specimens from 25 chronic liver disease patients and 28 normal healthy volunteers. The positive rate correlated with the TNM clinical stages, extrahepatic metastasis and portal vein carcinothrombosis (p<0.05). No correlation was found between tumor size, tumor number, differentiation, serum a-fetoprotein (AFP) and the positive rate. CONCLUSIONS: Our results indicate that a multimarker real-time Q-PCR assay with cancer-specific markers such as MAGE-1 and MAGE-3 in combination with a hepatocyte-specific AFP marker may be a promising diagnostic tool for monitoring hepato-cellular carcinoma patients with better sensitivity and specificity.

P53 and vascular endothelial growth factor expressions are two important indices for prognosis in gastric carcinoma.

AIM: This study examined the correlation between P53 and vascular endothelial growth factor (VEGF) expression together with tumour vascularity and investigated their clinical significance in the prognosis of gastric carcinoma. SUBJECTS AND METHODS: Ninety-five patients with gastric carcinoma who underwent curative surgical resection were studied using immunohistochemical staining. Correlation between the expression of p53, VEGF microvessel count (MVC) and various clinicopathologic factors were studied. RESULTS: No significant correlation was found between p53 expression and clinicopathologic factors. The rate of VEGF positivity was significantly higher in patients with haematogenous metastasis than in those without haematogenous metastasis. Both p53 and VEGF expression were associated with MVC. The MVC in p53 positive tumours was significantly higher than that in p53 negative tumours. Similarly, the same trend was seen between VEGF expression and MVC. The p53 and VEGF were co-expressed in 61 of 95 tumours (64.2%), and a significant (p < 0.01) association between p53 and VEGF expressions was demonstrated. The rate of VEGF positivity was significantly (p < 0.01) higher in the patients with disease recurrence than in those without recurrence, whereas no significant correlation was found between disease recurrence and the expression of p53. CONCLUSIONS: The p53 expression may play an important role in controlling angiogenesis by regulating VEGF expression and VEGF expression is associated closely with disease recurrence. In addition, both p53 and VEGF expression might be useful in indicating the prognosis in patients with gastric carcinoma.

MiR-451 Promotes Cell Proliferation and Metastasis in Pancreatic Cancer through Targeting CAB39.

Emerging evidence shows that microRNAs (miRNAs) play important roles in the regulation of various biological and pathologic processes in human cancers and the aberrant expression of miRNAs contributes to the tumor development. In this study, our findings indicate that miR-451 is significantly overexpressed in pancreatic cancer tissues and cell lines and elevated expression of miR-451 contributes to promoted cell viability (in vitro and in vivo). Moreover, overexpression of miR-451 is closely linked to poor prognosis and lymphatic metastasis. Inhibition of miR-451 dramatically suppresses cell viability and invasion, promotes cell apoptosis, and induces cell cycle arrest. Furthermore, miR-451 directly targets CAB39 and negatively regulates its expression and inhibition of CAB39 contributes to the promoted cell viability and invasion. Our findings improve our understanding of the function of miR-451 in the identification and therapy of pancreatic cancer.

[Identification of gastric cancer-related genes by multiple high throughput analysis and data mining].

OBJECTIVE: To investigate gastric cancer-related genes by combined multiple high throughput analysis and data mining, and to further identify gene markers that may be useful in the diagnosis and treatment of gastric cancer. METHODS: Data of expressed sequence tags (EST) and serial analysis of gene expression (SAGE) in Cancer Genome Anatomy Project (CGAP) were employed to analyze differential gene expression between normal and cancerous gastric epithelium,the obtained genes were further analyzed by virtual Northern blotting and compared with microarray data from Stanford Microarray Database (SMD). RESULTS: NCBI digital differential display (DDD), cDNA digital gene expression displayed (DGED) and SAGE DGED produced 165,286 and 181 differential expression genes.All these genes were analyzed by virtual Northern blotting and 45 genes were obtained. Comparing with microarray data, candidate genes were reduced to 12. Further RT-PCR analyses validated 4 genes, including ANXA1, MSMB, ANXA10 and PSCA, were differentially expressed in normal and cancerous gastric tissues. CONCLUSIONS: Combined multiple high throughput analysis and data mining is an effective strategy for identification of gastric cancer-related genes. Further analyses of these genes from data mining will provide biomarkers for the diagnosis and treatment of gastric cancer.

P53 and vascular endothelial growth factor expressions are two important indices for prognosis in gastric carcinoma / Las expresiones del P53 y del factor de crecimiento del endotelio vascular constituyen dos índices importantes para la prognosis del carcinoma gástrico

AIM: This study examined the correlation between P53 and vascular endothelial growth factor (VEGF) expression together with tumour vascularity and investigated their clinical significance in the prognosis of gastric carcinoma. SUBJECTS AND METHODS: Ninety-five patients with gastric carcinoma who underwent curative surgical resection were studied using immunohistochemical staining. Correlation between the expression of p53, VEGF, microvessel count (MVC) and various clinicopathologic factors were studied. RESULTS: No significant correlation was found between p53 expression and clinicopathologic factors. The rate of VEGF positivity was significantly higher in patients with haematogenous metastasis than in those without haematogenous metastasis. Both p53 and VEGF expression were associated with MVC. The MVC in p53 positive tumours was significantly higher than that in p53 negative tumours. Similarly, the same trend was seen between VEGF expression and MVC. The p53 and VEGF were co-expressed in 61 of 95 tumours (64.2%), and a significant (p < 0.01) association between p53 and VEGF expressions was demonstrated. The rate of VEGF positivity was significantly (p < 0.01) higher in the patients with disease recurrence than in those without recurrence, whereas no significant correlation was found between disease recurrence and the expression of p53. CONCLUSIONS: The p53 expression may play an important role in controlling angiogenesis by regulating VEGF expression and VEGF expression is associated closely with disease recurrence. In addition, both p53 and VEGF expression might be useful in indicating the prognosis in patients with gastric carcinoma.

OBJETIVO: Este estudio examinó la correlación entre el P53 y la expresión del factor de crecimiento del endotelio vascular (VEGF) junto con las vascularidad del tumor, e investigó su importancia clínica en la prognosis del carcinoma gástrico. SUJETOS Y MÉTODOS: Noventa y nueve pacientes con carcinoma gástrico que fueron sometidos a resección quirúrgica curativa, fueron estudiados usando teñido inmunohistoquímico. Se estudió la correlación entre la expresión de p53, VEGF, el conteo de microvasos (MVC) y varios factores clínico-patológicos. RESULTADOS: No se halló una correlación significativa entre la expresión de p53 y los factores clínico-patológicos. La tasa de positividad de VEGF, fue significativamente más alta en pacientes con metástasis hematogénica que en pacientes sin metástasis hematogénica. Tanto la expresión de p53 como la de VEGF estuvieron asociadas con el conteo MVC. El MVC en tumores p53 positivos fue significativamente más alto que en tumores p53 negativos. De manera similar, la misma tendencia se observó entre la expresión de VEGF y MVC. El p53 y el VEGA fueron co-expresados en 61 de 95 tumores (64.2%), y se demostró una asociación significativa (p < 0.01) entre las expresiones de p53 y VEGA. La tasa de positividad VEGA fue significativamente más alta (p < 0.01) en los pacientes con recurrencia de la enfermedad que en aquellos sin recurrencia, en tanto que no se halló una correlación significativa entre la recurrencia de la enfermedad y la expresión de p53. CONCLUSIONES: La expresión p53 puede desempeñar un importante papel en el control de la angiogénesis mediante la regulación de la expresión de VEGF y la expresión de VEGF está estrechamente asociada con la recurrencia de la enfermedad. Además, tanto la expresión de p53 como la de VEGF podrían ser útiles para indicar la prognosis de pacientes con carcinoma gástrico.