Correlation between Mnk2 protein expression and prognosis of patients with esophageal squamous cell carcinoma

AIM: To investigate the protein expression of mitogen-activated protein kinase-interacting kinase-2 (Mnk2) and its prognostic effect in the patients with resected esophageal squamous cell carcinoma (ESCC). METHODS: A total of 86 informative patients with surgically resected ESCC and 54 normal esophageal tissues were enrolled. Western blot and immunohistochemistry (IHC) were utilized to assess the protein expression of Mnk2, and its correlation with prognosis was statistically analyzed by the methods of Kaplan-Meier curve and Cox proportional hazard mode. RESULTS: The protein expression of Mnk2 was elevated in most of tumor tissues compared with the adjacent tissues. Clinicopathologic analysis showed that Mnk2 expression was significantly correlated with the TNM stage (P<0.05). Both disease-free survival (DFS) and overall survival (OS) of Mnk2 over-expression patients were shorter than those in Mnk2 negative expression group. Multivariate analysis confirmed that Mnk2 expression, as an independent and significant factor for both DFS and OS, predicted a poor prognosis of the patients with resected ESCC (P<0.05). CONCLUSION: The expression of Mnk2 was significantly related to the TNM stages, and might be a novel predictor for prognosis in ESCC.     

Chronic hypoxia enhances aggressiveness of MCF-7 breast cancer cells through EMT

AIM: To investigate the effects of chronic hypoxia on the aggressiveness of MCF-7, a human breast cancer cell line, and the underlying mechanisms.METHODS: MCF-7 cells were cultured under hypoxia (1% O₂, 5% CO₂ and 94% N₂) or control (95% O₂ and 5% CO₂) condition. The viability, proliferation, and invasion and migration abilities of the MCF-7 cells were determined by MTT assay, CCK-8 assay, cell counting, and cell invasion and migration assays. Anchorage-independent growth and the alteration of cellular polarization of the MCF-7 cells were tested by soft agar colony formation assay and Matrigel-3D culture assay, respectively. The effects of chronic hypoxia on the growth and metastasis of MCF-7 cells in vivo were investigated by xenograft in nude mice. The morphological changes of the MCF-7 cells were observed under an inverted microscope. Hypoxia-induced alterations in the levels of hypoxia inducible factor-1 (HIF-1) and phosphorylated glycogen synthase kinase-3β (p-GSK-3β) as well as epithelial-mesenchymal transition (EMT) molecules, such as E-cadherin, N-cadherin, vimentin, matrix metalloproteinase (MMP)-3 and MMP-9, were determined by Western blot.RESULTS: Chronic hypoxia significantly increased the viability, proliferation, and invasion and migration abilities of MCF-7 cells in vitro, enhanced the anchorage-independent growth, facilitated cellular polarization alteration in Matrigel-3D culture, and promoted cancer metastasis in vivo. Hypoxia up-regulated HIF-1, activated GSK-3β, down-regulated E-cadherin and increased the protein levels of N-cadherin, vimentin, MMP-3 and MMP-9. CONCLUSION: Chronic hypoxia enhances the aggressiveness of breast cancer cells probably through EMT.     

Dexmedetomidine attenuates acute alcoholic hepatic injury in mice

AIM: To investigate the effects of dexmedetomidine (DEX) on acute alcoholic hepatic injury in mice and to explore the possible mechanisms. METHODS: Kunming mice (n=50) were randomly divided into 5 groups (n=10): normal saline control (NS) group, acute alcoholic hepatic injury model (E) group, low-dose (10 μg/kg) DEX (E+L) group, medium-dose (50 μg/kg) DEX (E+M) group and high-dose (100 μg/kg) DEX (E+H) group. The animals were sacrificed at 6 h after gavage of alcohol or normal saline. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), malondialdehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD) were measured. The livers were removed for evaluation of histological characteristics and determining the content of tumor necrosis factor-α (TNF-α) amd interleukin-1β (IL-1β) in the liver tissues by ELISA. The expression levels of cytochrome P450 2E1 (CYP2E1) and nuclear factor-κB (NF-κB) in the liver tissues were evaluated by Western blot. RESULTS: Compared with NS group, the levels of ALT, AST and TG were obviously increased in E group, which were obviously decreased in E+M and E+H groups. Compared with NS group, the levels of TNF-α, IL-1β and MDA were obviously increase in E group, which were obviously decreased in E+M and E+H groups. Compared with NS group, the activity of SOD and the content of GSH were obviously decreased in E group, which were obviously increased in E+M and E+H groups. Compared with NS group, the expression of CYP2E1 and NF-κB was obviously increase in E group, which was obviously decreased in E+M and E+H groups. Compared with NS group, ethanol induced marked liver histological injury, which was less pronounced in E+M and E+H groups. CONCLUSION: DEX has a protective effect on mouse liver with acute alcoholic injury by the involvement in the processes of antioxidation and antiinflammation, and its mechanism may be associated with the inhibition of CYP2E1 and NF-κB expression.     

Effect of antisense locked nucleic acid blocking translation initiation region of c-myc exon 2 on apoptosis of hepatocellular carcinoma cells

AIM: To observe the effect of antisense locked nucleic acid (anti-LNA) blocking the translation initiation region of c-myc exon 2 on the viability and apoptosis of hepatocellular carcinoma cells.METHODS: The anti-LNA that was complementary to the translation initiation region of c-myc exon 2 was designed, synthesized, and introduced into the HepG2 cells by cationic liposome-mediated transfection. The mRNA and protein levels of c-Myc in the cells were determined by RT-PCR and Western blot. The change of cell apoptosis was analyzed by flow cytometry, and the toxicity of anti-LNA to the cells was detected by MTT assay.RESULTS: Five days after transfection, the mRNA level of c-Myc in anti-LNA group was 0.335±0.016, and the protein level was 0.448±0.037, significantly lower than those in control group (both P<0.05). The ratio of apoptotic cells in anti-LNA group was 32%±6%, which was higher than that in control group (P<0.05).CONCLUSION: Antisense locked nucleic acid targeting at the translation initiation region of c-myc exon 2 shows strong inhibitory effects on the apoptosis of hepatocellular carcinoma cells.     

Effect of tetrandrine on radiosensitivity of nasopharyngeal carcinoma cells

AIM: To investigate the mechanism of the radiosensitizing effect of maximum non-cytotoxic doses of tetrandrine (Tet) on nasopharyngeal carcinoma cell lines CNE1 and CNE2.METHODS: The cells were treated with ma-ximum non-cytotoxic doses of Tet (for CNE1 cells at 1.5 μmol/L and for CNE2 cells at 1.8 μmol/L), irradiation at 4 Gy, or combination of irradiation and maximum non-cytotoxic doses of Tet. The cell cycle distribution was analyzed by flow cytometry. The protein levels of γ-H2AX, cleaved caspase-3, p-CDC25C, CDK1, p-CDK1, cyclin B1, ERK and p-ERK were determined by Western blot.RESULTS: The expression of γ-H2AX was increased in CNE1 cells and CNE2 cells after combined treatment with irradiation and maximum non-cytotoxic doses of Tet. The percentages of CNE1 cells and CNE2 cells at G₂/M phase in irradiation group were (18.09±0.42)% and (18.48±1.32)%, respectively, which were decreased to (15.88±1.04)% and (13.80±0.82)% in combined treatment group, respectively (P<0.05). Combined treatment enhanced the increase in the protein level of cleaved caspase-3 caused by irradiation. The protein levels of p-CDC25C and p-CDK1 were increased in a dose-dependent manner by Tet treatment (P<0.05), while the expression of CDK1 showed no difference among different doses of Tet treatments. The protein levels of p-CDC25C, p-CDK1 and CDK1 showed no difference after the treatment with maximum non-cytotoxic doses of Tet. The combined treatment with irradiation and the maximum non-cytotoxic doses of Tet decreased the protein levels of p-CDC25C and p-CDK1 (P<0.05), increased the expression of cyclin B1, and had no influence on the expression of CDK1 (P<0.05). The combined treatment resulted in an increase in the protein level of p-ERK1 (P<0.05).CONCLUSION: The maximum non-cytotoxic doses of Tet enhance the DNA damage and apoptosis in CNE1 cells and CNE2 cells caused by irradiation, and the mechanism might be associated with ending of G₂/M arrest via activation of ERK/CDC25C/CDK1/cyclin B1 pathways.     

山核桃果皮提取物抗肿瘤活性研究

[目的]研究山核桃果皮提取物抗肿瘤活性。[方法]采用提取及萃取过程制备山核桃果皮提取物分离化合物,选取人肝癌SMMC-7721细胞及人宫颈癌Hela细胞为模型,测定山核桃果皮提取物对肿瘤细胞的抑制作用。[结果]从山核桃果皮所提取出的化合物具有一定的体外增殖抑制作用。[结论]山核桃果皮提取物是一种安全有效的抗肿瘤活性物质。     

饲粮生物素水平对团头鲂幼鱼肠道消化酶活性、胴体组成及肝脏抗氧化能力的影响

选择初重为201±001 g团头鲂幼鱼720尾,分别饲喂生物素添加量为0,002,005,016,062和249 mg·kg-1等氮等能的纯化饲料,研究饲料中添加生物素对团头鲂幼鱼肠道消化酶活力、胴体组成及肝脏抗氧化的影响。结果表明: 团头鲂幼鱼肠道脂肪酶和蛋白酶活力随生物素含量的升高呈现先增加后下降的变化,它们分别在生物素含量为062和016 mg·kg-1时达到最大值。饲料生物素含量为016 mg·kg-1组的团头鲂幼鱼与对照组相比具有较低的水分和较高的粗脂肪、粗蛋白含量,差异显著(P<005)。饲料中生物素含量从0上升至005 mg·kg-1时,GSH含量,GSH Px、SOD活性显著升高,差异显著(P<005);当其含量进一步升高至249 mg·kg-1时,三者均无显著变化(P>005)。综上所述,饲料中添加适宜浓度生物素可以提高团头鲂幼鱼肠道脂肪、蛋白质的消化能力,胴体粗脂肪和粗蛋白含量及肝脏抗氧化能力。     

Effect of dietary betaine levels on growth performance and hepatic intermediary metabolism of GIFT strain of Nile tilapia Oreochromis niloticus reared in freshwater

An 8‐week feeding experiment was conducted to determine the effect of dietary betaine levels on the growth performance and hepatic intermediary metabolism of genetically improved farmed tilapia (GIFT) strain of Nile tilapia Oreochromis niloticus (mean initial body weight: 78.3 ± 1.3 g, means ± SD). Six practical diets were formulated with the incorporation of betaine at the levels of 0 (control), 5, 10, 15, 20 and 25 g kg⁻¹. Survival showed no significant differences among the treatments (P > 0.05). The highest and lowest weight gain (WG) and specific growth rate (SGR) were observed for fish fed the diets containing 5 and 0 g kg⁻¹ (control) betaine, respectively. Feed intake showed similar trend with WG and SGR. In contrast, feed conversion ratio was the lowest when dietary betaine level was 5 g kg⁻¹. In general, dietary betaine supplementation showed no significant effect on hepatic composition of tilapia. Condition factor and viscerosomatic index tended to increase with increasing dietary betaine levels from 0 to 5 g kg⁻¹ and then decline when dietary betaine levels further increased from 5 to 25 g kg⁻¹. In contrast, hepatosomatic index declined with increasing dietary betaine levels (P < 0.05). Dietary betaine levels significantly influenced several hepatic enzymatic activities, including succinate dehydrogenase, lactate dehydrogenase, malic dehydrogenase, lipoprotein lipase and hepatic lipase, suggesting that dietary betaine addition had significant effects on nutrient metabolism in the liver. Based on the second‐order polynomial regression analysis of WG, 12.5 g kg⁻¹ of dietary betaine level seemed optimal for genetically improved farmed tilapia strain of O. niloticus.     

Effects of dietary carbohydrate sources on the growth performance and hepatic carbohydrate metabolic enzyme activities of juvenile cobia (Rachycentron canadum Linnaeus.)

An 8-week feeding trial was conducted to evaluate the effect of dietary carbohydrate sources on the growth performance and hepatic carbohydrate metabolic enzyme activities of juvenile cobia. Six experimental diets were formulated to contain 20% glucose, sucrose, maltose, dextrin, corn starch and wheat starch respectively. The results indicated that fish fed the wheat starch and dextrin diets showed significantly better weight gain, specific growth rate and protein efficiency ratio compared with those fed the other diets. However, fish fed the glucose diet had a significantly lower survival and condition factor than those fed the other diets. There were significant differences in the total plasma glucose and triglyceride concentration in fish fed diets with different dietary carbohydrate sources. Haematocrit, haemoglobin, red blood cell and leucocytes were significantly affected by the dietary carbohydrate sources. The activities of glucose-6-phosphate dehydrogenase (G6PD), 6-phosphofructokinase (PFK) and fructose-1,6-bisphosphatase (FBPase) were significantly affected by the dietary carbohydrate sources, while fish fed the glucose diet showed higher G6PD, PFK and FBPase activities than those fed the other diets. These data indicated that dextrin and wheat starch were the most optimal carbohydrate sources for juvenile cobia.