MicroRNA-375 targets PAX6 and inhibits the viability, migration and invasion of human breast cancer MCF-7 cells.

[This retracts the article DOI: 10.3892/etm.2017.4593.].

Targeting TRPV1 on cellular plasticity regulated by Ovol 2 and Zeb 1 in hepatocellular carcinoma.

The landscape of cellular plasticity and sources with relevant niche signals in hepatocellular carcinoma is still obscure. Transient receptor potential vanilloid 1 (TRPV1), a non-selective cation channel, is involved in a variety of malignancies and overexpressed in hepatocellular carcinoma (HCC). We have investigated the role of TRPV1 in HCC from different angles by various experimental techniques, such as in vivo and in vitro experiments, and by bioinformatics analysis of data from genetic models induced by diethylnitrosamine (DEN), mice samples and human HCC samples. We find that TRPV1 knockout promotes to hepatocarcinogenesis and deconstructs the portal triad adjacent to tumor border that is contributed by originations of tumor initiating cells and biliary cells. Epithelial to mesenchymal transition (EMT) is involved and transcription factors Ovol2 and Zeb1 coordinated with Sox 10 drive gene expression in the event which is also confirmed by the expression of these proteins in human HCC samples. Treatment with TRPV1 agonist Capsaicin inhibits the growth of HCC cells in xenograft models. Our findings demonstrate that TRPV1 is a potential therapeutic target in human HCC and exerts effects on cellular plasticity with modulation of Ovol2, Zeb1 and Sox10.

MicroRNA-375 targets PAX6 and inhibits the viability, migration and invasion of human breast cancer MCF-7 cells.

MicroRNAs (miRs) are a type of small non-coding RNA that serve crucial roles in the development and progression of breast cancer. However, the exact role and underlying molecular mechanism of miR-375 in mediating the growth and metastasis of breast cancer remains unknown. In the present study, reverse transcription-quantitative polymerase chain reaction and western blot analysis were conducted to examine RNA and protein expression. A luciferase reporter assay was performed to determine the association between miR-375 and paired box 6 (PAX6). The results of the current study indicate that the expression of miR-375 was reduced in breast cancer tissues compared with matched adjacent normal tissues. Transfection with miR-375 mimics led to a significant increase in levels of miR-375 in human breast cancer Michigan Cancer Foundation (MCF)-7 cells (P<0.05). The increase in miR-375 expression caused a significant decrease in the viability, migration and invasion of MCF-7 cells (P<0.05), accompanied by a reduced expression of matrix metalloproteinase (MMP) 2 and MMP9 proteins. Luciferase reporter assay identified PAX6 as a novel target of miR-375 and miR-375 in turn, negatively regulated the protein expression of PAX6 in MCF-7 cells. By contrast, overexpression of PAX6 led to a significant increase in MCF-7 cell viability (P<0.01) but did not affect the migration and invasion of MCF-7 cells, suggesting that the inhibitory effect of miR-375 on MCF-7 cell viability may be occurring, in part, via the direct targeting of PAX6.

AEG-1 is associated with hypoxia-induced hepatocellular carcinoma chemoresistance via regulating PI3K/AKT/HIF-1alpha/MDR-1 pathway.

Hypoxia is a common characteristic of hepatocellular carcinoma (HCC) associated with reduced response to chemotherapy, thus increasing the probability of tumor recurrence. Astrocyte elevated gene-1 (AEG-1) has been involved in a wide array of cancer progression including proliferation, chemoresistance, angiogenesis and metastasis, but its effect on HCC chemoresistance induced by hypoxia is unclear. In this study, expression of AEG-1 and multiple drug resistance (MDR-1) were examined in HCC using immunohistochemical staining and RT-PCR. Furthermore, their expression levels were detected in HCC HepG2 cells in normoxia or hypoxia via RT-PCR and Western blot assays. Specific shRNAs were used to silence AEG-1 expression in HepG2 cells. Results showed AEG-1 and MDR-1 expression were higher in HCC tissues than in adjacent normal tissues. Incubation of HepG2 cells in hypoxia increased expression of AEG-1 and MDR-1, compared to incubation in normoxia. Exposure to hypoxia blunted sensitivity of HepG2 cells to Adriamycin, 5-fluorouracil and cis-platinum, as evidenced by modest alterations in cell viability and apoptosis rate, however the sensitivity was elevated with AEG-1 knockdown. PI3K/AKT/HIF-1/MDR-1 pathway was attenuated following AEG-1 knockdown in hypoxia. Based on these data, it was suggested that AEG-1 is associated with hypoxia-induced hepatocellular carcinoma chemoresistance via regulating PI3K/AKT/HIF-1/MDR-1 pathway. This study uncovered a novel potential target for development of an effective therapy against hypoxia-induced HCC chemoresistance.

Numb downregulation suppresses cell growth and is associated with a poor prognosis of human hepatocellular carcinoma.

Numb, an endocytic adaptor, is a known cell fate determinant that participates in asymmetric cell division. The present study aimed to explore the potential roles of Numb in hepatocarcinogenesis. Numb expression was investigated in hepatocellular carcinomas (HCC) with reverse transcription­quantitative polymerase chain reaction and immunohistochemical examination; its association with the prognosis of HCC patients was analyzed. In addition, the effects of Numb deletion on proliferation of HCC cells and its relevant molecules were evaluated in Huh7 and HepG2 cells. Numb overexpression was observed in 62% of adjacent non­tumor tissues and 46% of tumor tissues. Overexpression of Numb in HCC was associated with histological grade, portal vein invasion and the number of tumors (P=0.001, 0.022 and 0.034 respectively). Multivariate analysis revealed that Numb expression was an independent prognostic indicator of HCC patients. Methylation of the Numb promoter contributed to hepatocarcinogenesis. In vitro assays demonstrated that Numb silencing resulted in inhibition of cell proliferation, induction of apoptosis, downregulation of cyclin­dependent protein kinase 4 (CDK4) and S­phase kinase­associated protein 2 (SKP2), and upregulation of Bcl­2 homologous antagonist/killer (BAK) and cyclin­dependent kinase inhibitor 1 (p21). The present study suggests that downregulation of Numb inhibits colony formation and cell proliferation, induces apoptosis of HCC cells and independently predicts the poor prognosis of HCC patients. Thus, Numb has a potential role in the development and progression of HCC.

[Application of liver hanging maneuver in anterior approach for isolated complete liver caudate lobectomy].

OBJECTIVE: To explore the technique and effect of liver hanging maneuver in anterior approach for isolated complete liver caudate lobectomy. METHODS: We recruited 17 patients with liver caudate lobe tumor (13 primary hepatocellular carcinoma, 3 cholangiocarcinoma and 1 liver metastasis from colorectal cancer). Isolated complete caudate lobectomy with liver hanging maneuver was performed in 17 patients. RESULTS: All 17 patients were successfully received the above-mentioned operation. The operative time was 166-427 (211.5 ± 20.1) min and the intraoperative blood loss was 372-1 208 (472.7 ± 83.6) mL. There was no operative death. The survival rates of follow up for 1, 3 and 5 years were 76.5%, 52.9% and 23.5%, respectively. CONCLUSION: Liver hanging maneuver for isolated complete resection of the caudate lobe is an ideal approach for liver neoplasms resection.

Cytotoxicity of hollow silica nanoparticles loaded with photosensitizes on huh-7 cells.

To observe the cytotoxic effect of the photodynamic therapy mediated by the traditional photosensitizer polyhematoporphyrin (C(34)H(38)N(4)NaO(5), Photosan-II Photosan-II was loaded into HSNP by one-step wet chemical, PS) and hollow silica nanoparticles (HSNP) loaded PS on Huh-7 cells and compare the cytotoxic effects. -based synthetic route. The cellular viability was determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Apoptotic and necrotic cells were measured by flow cytometry. The suitable drug concentrations of free PS and HSNP loaded PS on Huh-7 cells were 20mg/L and 5mg/L respectively, and the suitable incubation time were 4 h and 2 h respectively. Under the same drug concentration, the survival rates of free PS and HSNP loaded PS were 62.46%±1.93% and 6.54%±1.24% <. Under the same drug concentration and incubation time, the total rates of apoptosis necrosis caused by free PS and HSNP loaded PS mediated PDT were respectively 22.00% ± 2.24% and 87.23% ± 2.1% <. PS-loaded HSNP mediated PDT can inhibit proliferation of cancer cells and induce apoptosis more quickly and effectively. As the loading system of PS, HSNP can make the photosensitizer have stronger solubility and drug concentration efficiency, which can significantly improve the therapeutic effect of PDT.

microRNA-200a inhibits cell proliferation by targeting mitochondrial transcription factor A in breast cancer.

microRNAs (miRNAs) are endogenous 19-25 nucleotide noncoding single-stranded RNAs that regulate gene expression by blocking the translation or decreasing the stability of mRNAs. In this study, we showed that miR-200a expression levels were decreased while mitochondrial transcription factor A (TFAM) mRNA expression levels were increased in breast cancer (BC) tissues and cell lines, and identified TFAM as a novel direct target of miR-200a. Overexpression of miR-200a suppressed TFAM protein expression, mtDNA copy number, and attenuated cell proliferation. Forced expression of TFAM can partly rescue the inhibitory effect of miR-200a in the cells. Taken together, these findings will shed light on the role and mechanism of miR-200a in regulating BC cells growth and mtDNA copy number via miR-200a/TFAM axis, and miR-200a may serve as a potential therapeutic target in BC in the future.

Ferroportin in the progression and prognosis of hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is the fifth most common malignant tumor in men and the seventh in women and understanding the molecular mechanisms of HCC and establishing more effective therapies are critical and urgent issues. Our objective was to study the expression of ferroportin in hepatocellular carcinoma (HCC) tissue samples and the relationship between ferroportin expression and HCC characteristics. METHODS: Sixty HCC tissues and their corresponding para-cancer liver tissues (PCLT) were obtained from sixty HCC patients who had undergone hepatectomy in the Second Xiangya Hospital of Central South University. Ten normal liver tissue samples were also obtained as a control. Immunohistochemistry (IHC) was performed to analyze the ferroportin expression in HCC, and the relationship between ferroportin expression and HCC clinical pathological characteristics also was analyzed. For the evaluation of IHC results, the comprehensive scoring criteria were met according to the staining intensity and the number of positive staining cells. Western blotting was performed to detect the expression level of ferroportin in HCC cell lines. RESULTS: Ferroportin expression in HCC tissue was significantly lower compared to PCLT and normal liver tissue (P <0.05). Moreover, ferroportin expression was related to liver cancer cell de-differentiation, the severity degree in TNM staging, Edmondson-Steiner grading, intrahepatic metastasis and portal vein invasion. In addition, high expression of ferroportin was observed in normal human liver cell lines L02 and HL7702, whereas weak positive expression and even negative expression of ferroportin were observed in HCC cell lines FOCUS, MHCC-97H, HepG2 and SMMC-7721. Furthermore, among the four kinds of HCC cell lines, the expression level of ferroportin was the lowest in MHCC-97H cells. CONCLUSIONS: Ferroportin expression level declines along with the progression of liver cancer, suggesting that the reduction of ferroportin may serve as an important marker for poor HCC prognosis and as a new therapeutic target.

Peliosis hepatis mimicking cancer: A case report.

Peliosis hepatis (PH) is a rare condition characterized by the presence of blood-filled cavities within the liver. The etiology of PH remains unknown, but it has been reported to be associated with infections or malignancies. However, the cause of PH is not clear in 20-50% of patients. The current study presents the case of a 19-year-old male who presented with right upper quadrant pain that had lasted for three days. The patient was a student with no previous medical history. Contrast-enhanced computer tomography (CT) and ultrasonography showed a neoplasm in the right liver and a diagnosis of primary liver cancer was made due to the manifestation of the disease and the results of physical tests. The individual was treated successfully with an irregular right hemihepatectomy and was in good health at 6-months post-surgery. A tissue specimen was obtained and determined to be PH by pathological examination and immunohistochemistry analysis. Consequently, a diagnosis of PH must be considered in cases like this.

[Influence of BRAF interference on SW579 cell lines in thyroid cancer].

OBJECTIVE: To determine the influence of v-raf murine sarcoma viral oncogene homolog B1 (BRAF) interference on SW579 cell lines in thyroid cancer. METHODS: We designed 2 pairs of siRNA interference sequences, transfected them into SW579 cell line with liposome, and detected the interference with RT-PCR method. For successfully interfered cell lines, the changes in cell proliferation, cell cycle, and the expression of related proteins in mitogen-activated protein signal-regulated kinase /extracellular signal-regulated kinase (MEK/ ERK) signal pathway were detected. RESULTS: After 2 pairs of siRNA transfection, the expressions of BRAF mRNA and protein of SW579 cell lines were significantly inhibited (P<0.01). The proliferation was inhibited, the cell cycle was changed, G1/S phase increased, and MEK/ERK signal pathway was inhibited. CONCLUSION: Inhibition of growth and proliferation of SW579 cell lines by BRAF may be functioned by de-activating MEK/ERK signal pathway.

Inhibitory effect of geraniol in combination with gemcitabine on proliferation of BXPC-3 human pancreatic cancer cells.

OBJECTIVE: To investigate the inhibitory effect of geraniol alone, or in combination with gemcitabine, on the proliferation of BXPC-3 pancreatic cancer cells. METHODS: BXPC-3 cells were treated under different conditions: with geraniol at 10, 20, 40, 80 and 160 µmol/l each for 24 h, 48 h or 72 h; with 20 µmol/l geraniol for 24 h or 0 h before 20 µmol/l gemcitabine for 24 h; with 20 µmol/l geraniol for 24 h, 48 h and 72 h following 20 µmol/l gemcitabine for 24 h; or with 20 µmol/l gemcitabine alone as a control. Cell proliferation was assessed and changes in cell morphology were assessed by light and fluorescence microscopy. Apoptosis was detected using flow cytometry. RESULTS: Geraniol inhibited BXPC-3 cell proliferation in a time- and dosa-dependent manner. Geraniol alone or combined with gemcitabine induced BXPC-3 cell apoptosis. BXPC-3 inhibition rates with combined treatment were 55.24%, 50.69%, 49.83%, 41.85% and 45.27% following treatment with 20 µmol/l geraniol for 24 h or 0 h before 20 µmol/l gemcitabine for 24 h, or 20 µmol/l geraniol for 24 h, 48 h and 72 h, following 20 µmol/l gemcitabine for 24 h, respectively. CONCLUSION: Geraniol inhibited the proliferation of BXPC-3 cells. Geraniol significantly increased the antiproliferative and apoptosis-inducing effects of gemcitabine on BXPC-3 cells. Maximum inhibition of BXPC-3 cells was achieved with geraniol treatment for 24 h before gemcitabine treatment.

Optimization extraction of Ganoderma lucidum polysaccharides and its immunity and antioxidant activities.

Extraction of Ganoderma lucidum polysaccharides (GLP) was optimized by response surface method (RSM). By running the optimization program with design expert within the experimental range investigated, the following optimum values were obtained: extraction time 230 min; extraction temperature 95 °C, and extraction number 5. The predicted polysaccharides production was 1.45%. Results showed that GLP significantly reduced the levels of serum IL-6 and TNF-α levels and increased the levels of serum IL-2, IL-4 and IL-10 in GLP-treated rats compared to gastric cancer model rats. In addition, administration of Ganoderma lucidum polysaccharides to GLP-treated group of rats improved the levels of serum and gastric tissue SOD, CAT and GSH-Px toward the control values in a dose-dependent manner. These findings show that GLP can enhance immunity and antioxidant activities in gastric cancer rats.

Effect of the PA-MSHA vaccine on septic serum-induced inflammatory response.

Sepsis is defined as a complex clinical syndrome caused by a serious infection followed by an amplified and deregulated inflammatory response. The complex syndrome is associated with a high rate of morbidity and mortality, despite substantial clinical advances. A vaccine derived from the outer membrane proteins of the Gram-negative bacteria Pseudomonas aeruginosa (PA-MSHA) has been demonstrated to exhibit immune modulatory properties. In the present study, the effect of the PA-MSHA vaccine on the inflammatory response induced by serum from septic patients in peripheral blood mononuclear cells was determined. It was observed that PA-MSHA pretreatment inhibits the production of septic serum-induced tumor necrosis factor-α. In addition, PA-MSHA treatment increases interleukin-10 levels and promotes the generation of CD4+CD25+Foxp3+ T cells. Thus, the results of the current study provide mechanistic insight relevant to the potential application of PA-MSHA in the treatment of sepsis.

Adiponectin (ADIPOQ) rs2241766 G/T polymorphism is associated with risk of cancer: evidence from a meta-analysis.

Associations between adiponectin (ADIPOQ) genetic polymorphisms (rs2241766 G/T and rs266729 G/C) and cancer risk have been extensively studied in the past decade, while conflicting results were reported. Therefore, this study would explore the associations by using a meta-analysis. The databases of Medline, Embase, and Wangfang were retrieved, and the latest updated time was 1 August 2012. Effect sizes of odds ratio and 95 % confidence interval (OR and 95 % CI) were calculated by using a fixed- or random-effect model. A total of 12 studies with 10,368 participants were identified for association between ADIPOQ rs2241766 G/T and risk of cancer, and ten studies with 12,665 participants were for association between ADIPOQ rs266729 G/C and risk of cancer. Overall combined analyses indicated that neither ADIPOQ rs2241766 G/T nor rs266729 G/C was associated with risk of cancer incidence (OR (95 % CI), 0.89 (0.61-1.30) for GG vs. TT and 0.94 (0.83-1.06) for G carriers vs. T carriers for rs2241766 G/T; 0.99 (0.85-1.16) for GG vs. CC and 0.96 (0.87-1.06) for G carriers vs. C carriers for rs266729 G/C). When stratified analyses were conducted according to the participants' ethnicity, sources of controls, types of cancer, and sample size, we found that G allele of ADIPOQ rs2241766 G/T was significantly associated with decreased risk of cancer based on population-based case-control studies (OR (95 % CI), 0.65 (0.50-0.85) for GG vs. TT and 0.88 (0.79-0.98) for G carriers vs. T carriers). In contrast, there was no association between rs266729 G/C polymorphism and risk of cancer when subgroup analyses were conducted. In summary, this meta-analysis indicated that ADIPOQ rs2241766 G/T rather than rs266729 G/C polymorphism was closely associated with risk of cancer development.

Extraction and bioactivity of polygonatum polysaccharides.

The present study is to explore the optimal extraction parameters and liver protective effect of the polygonatum polysaccharides in vivo. The order of factor effects on polysaccharides production was found to be extraction time (min, A)>ratio of solvent to solid (C)>extraction temperature (°C, B)>extraction number (D). The results show that the effects of extraction time (min, A) and ratio of solvent to solid (C) were more significant than those of the other factors. Optimal extraction parameters were as followings: extraction time 120 min, extraction temperature 100 °C, ratio of solvent to solid 5, and extraction number 4. Polygonatum polysaccharides was administered orally at doses of 150, 300 and 450 mg/(kg day) to carbon tetrachloride (CCl(4))-treated rats. Results showed that administration of polygonatum polysaccharides could increase rats' final body weight, liver antioxidant enzymes activities (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and glutathione reductase (GR)), decrease serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities and liver malondialdehyde (MDA) level. The liver sections obtained from animals supplemented with polygonatum polysaccharides extract demonstrated reduced pathological damages, supporting that polygonatum polysaccharides extract could effectively decrease the toxicity of CCl(4). It can be concluded that polygonatum polysaccharides treatment may prevent CCl(4)-induced liver oxidative injury in experimental rats.

Situs inversus totalis with carcinoma of gastric cardia: a case report.

Situs inversus is an uncommon anomaly with rare incidence. Some cases of situs inversus totalis have been described with different types of associations. Here we report a case of situs inversus with carcinoma of the gastric cardia.

Anti-proliferative effects of anandamide in human hepatocellular carcinoma cells.

In our previous study, we reported that the cannabinoid receptors CB1 and CB2 are overexpressed in human hepatocellular carcinoma (HCC) tissues. Recently, the antitumor potential of the endogenous cannabinoid anandamide (AEA) has also been addressed. The present study was conducted to investigate the anti-proliferative effects of AEA in HCC cells. The human HCC cell line Huh7 was used. Cell proliferation was measured by MTT assay and flow cytometry. Apoptotic analysis was investigated by TUNEL assay. Real-time PCR and western blot analysis were used to analyze the expression of relevant molecules. The results of this study demonstrated that AEA inhibited the proliferation of Huh7 cells, resulted in G1 cell cycle arrest and induced apoptosis. Furthermore, downregulation of CDK4 and upregulation of p21 and Bak by AEA were observed. This study defines the anti-proliferative effects of anandamide in HCC cells and suggests that AEA has therapeutic potential in the management of HCC patients.

[Expression of BRAF and its extracellular signal-regulated kinase 1/2 signal pathway in papillary thyroid cancer].

OBJECTIVE: To determine the association between activity of BRAF and mitogen-activated protein/ extracellular signal-regulated kinase kinase (MEK) / extracellular signal-regulated kinase (ERK) signal pathway in papillary thyroid cancer and its mechanism. METHODS: We collected the clinical data and blood samples from 73 cases of papillary thyroid cancer and another 16 cases of benign thyroid gland tumor, and detected the expression of rat sarcoma (RAS), BRAF, MEK1/2, and ERK1/2 in all tumor specimens and benign thyroid tissues with immunohistochemistry and Western blot. RESULTS: The expression of RAS, BRAF, pMEK1/2, and pERK1/2 protein in papillary thyroid cancer tissues was higher than those in the benign thyroid tissues(P<0.05 or P<0.01). The expression of RAS, BRAF, MEK1/2, and ERK1/2 was associated with the tumor size, the lymph node metastasis, and the clinical stage of papillary thyroid cancer(P<0.05 or P<0.01). CONCLUSION: The expression of RAS, BRAF, pMEK1/2, and pERK1/2 is associated with the pathogenesis, the lymph node metastasis, and the clinical stage of papillary thyroid cancer. The MEK/ERK signaling pathway may be activated by BRAF in papillary thyroid cancer.