The Clinical and Prognostic Implications of Pluripotent Stem Cell Markers Expression and Their Correlation with the WNT signal pathway in Hepatocellular Carcinoma

Objectives: This study aimed to investigate the expression of SOX2, SOX9, p53, and β-catenin in hepatocellular carcinoma (HCC) and their correlation with clinicopathological parameters of prognostic importance. Materials and Methods: Seventy-five patients were enrolled in this study. All patients had full clinical and follow-up data and available paraffin blocks. Immunohistochemical analysis was performed and correlated with clinicopathological factors and patient survival. Results: We detected the positive expression of SOX2, SOX9, p53, and β-catenin in 76%, 50.7%, 50.7%, and 77.9% of HCC specimens respectively. All studied markers showed a significant increase in the expression in tumor tissue specimens compared to non-tumor tissue. Both SOX2 and SOX9 expressions were significantly associated with adverse prognostic factors in HCC. Significant positive correlations were found between SOX2 and SOX9 and both p53 and β-catenin expression (r= 0.528, 0.485 and; r = 0.253, 0.327, respectively; p < 0.0001 for both of them). Regarding survival, we found that HCC patients with positive SOX2 and SOX9 expressions had significantly shorter overall survival (p=0.0001, each). Additionally, larger tumor size, tumor grade, high stage, tumor multiplicity, presence of cirrhosis, tumor necrosis, high p53 expression, and positive β-catenin expression were independent predictors of worse survival. A multivariate Cox analysis revealed that tumor grade, stage, p53, and SOX2 expression were independent predictors of unfavorable prognosis in overall survival (p=0.0001, p=0.0001,p=0.033; and p=0.003, respectively). Conclusions: Our findings might provide an insight into SOX2 and SOX9’s role in HCC and suggest that SOX2 might be targeted for HCC therapy.     

大鼠非特异性睾丸炎模型的建立及生精上皮的变化

目的:探讨大鼠生精上皮在非特异性炎症状态下的变化。方法:20只成年W istar雄性大鼠随机分为2组(对照组和实验组),对照组腹腔注射生理盐水(1 m l/kg),实验组注射细菌内毒素(LPS),剂量为1 mg/kg,两组均隔1 d注射1次,距第1次注射10 d后取材。HE染色观察睾丸组织的病理改变,免疫组化方法探讨生精上皮内增殖细胞核抗原(PCNA)、α连接素的变化。结果:实验组睾丸组织有明显炎性改变;生精上皮内PCNA表达量显著降低:每个生精小管(仅精原细胞着色的小管)阳性细胞个数为(59±5)个,比对照组明显减少(P<0.01),而这种小管占总管数的比例显著升高(P<0.01),为0.673±0.054;实验组大鼠睾丸内α连接素表达量亦减少(P<0.01),阳性反应颗粒平均光密度为0.150±0.014。结论:睾丸炎症时精原细胞增殖和生精上皮内细胞间粘附力度均减弱,这可能是睾丸炎导致男性不育的原因之一。     

Expression of β-catenin in normal breast tissue and breast carcinoma: a comparative study with epithelial cadherin and α-catenin

Expression of β-catenin was investigated in normal breast tissue and 66 breast carcinomas in conjunction with expression of epithelial cadherin (E-CD) and α-catenin. In normal mammary ducts and acini, intense β-catenin immunoreactivity was present at the basolateral surfaces of luminal epithelium and weak immunoreactivity was observed at the lateral borders of myoepithelial cells. No β-catenin was revealed at the myoepithelial basal surface. The intercellular expression of β-catenin, as well as of E-CD and α-catenin, was also observed in carcinoma tissues with varying staining intensity. Almost all of 10 intraductal carcinomas and approximately 70% of 41 invasive ductal carcinomas expressed the three molecules at the same level as in normal glands, whereas approximately 80% of 13 invasive lobular carcinomas showed severe deficiency of them. Two lobular carcinomas in situ showed complete absence of all of the proteins. Some of these findings were confirmed biochemically by immunoblotting analysis. In invasive ductal carcinomas, α-catenin was reduced more frequently in diffuse than in solid type tumours, whereas the level of expression of β-catenin and E-CD was unchanged between them. No correlation was present between reduced expression of the adhesion molecules and lymph node metastasis.     

A subset of high-grade pulmonary neuroendocrine carcinomas shows up-regulation of matrix metalloproteinase-7 associated with nuclear β-catenin immunoreactivity,independent of EGFR and HER-2 gene amplification or expression

Nuclear translocation of β-catenin has been correlated with epidermal growth factor receptor (EGFR) overexpression/activation in nonsmall cell lung cancer. Less is known on β-catenin transactivation in high-grade pulmonary neuroendocrine tumors and on the status of β-catenin activating EGFR and human epidermal growth factor receptor 2 (HER-2) or β-catenin target genes cyclin D1 and matrix metalloproteinase-7 (MMP-7). β-catenin immunoreactivity was evaluated in 51 large-cell neuroendocrine carcinomas (LCNEC) and 45 small-cell lung carcinomas (SCLC). Nineteen cases were assessed for β-catenin gene exon 3 mutations, expression of MMP-7, and expression/gene amplification of EGFR, HER-2, and cyclin D1. β-catenin was expressed in all 96 high-grade neuroendocrine tumors, the vast majority (94%) showing >50% immunopositive cells. A disarrayed immunoreactivity, however, was commonly encountered consisting in variably altered membrane-associated patterns of staining along with progressive accumulation of cytoplasmic immunoreactivity. In LCNEC, but not in SCLC, the disarrayed patterns correlated with EGFR and HER-2 protein expression. β-catenin nuclear accumulation was found in nine tumors, including seven LCNEC and two SCLC, and was always associated with disarrayed immunoreactivity and increased MMP-7, but not cyclin D1 expression. These cases, however, did not show β-catenin gene mutations or EGFR and HER-2 gene amplification or expression. No association was found between nuclear β-catenin and any clinicopathological variable including patients' survival. The subcellular compartmentalization of β-catenin is profoundly altered in high-grade pulmonary neuroendocrine tumors. A minor subset of these tumors shows β-catenin nuclear accumulation in association with increased expression of MMP-7, but not of cyclin D1, independent of EGFR and HER-2 gene amplification or expression. The authors have no significant financial or other relationship with the manufacturers of any commercial products or commercial services presented in this paper     

Aberrant expression of beta-catenin and mutation of exon 3 of the beta-catenin gene in renal and urothelial carcinomas

The present study attempted to clarify the significance of aberrant expression of beta-catenin protein and mutation of exon 3 of the beta-catenin gene in renal and urothelial carcinogenesis. beta-Catenin expression was examined immunohistochemically and mutation of the beta-catenin gene was analyzed by polymerase chain reaction-single strand conformation polymorphism (SSCP) and direct sequencing. beta-Catenin immunoreactivity was observed at the cell membrane in all 30 renal cell carcinomas (RCC) examined, and no RCC showed a mobility-shifted SSCP band. Of 46 transitional cell carcinomas (TCC) examined, there was reduced expression of beta-catenin, as compared with its expression in non-cancerous transitional epithelium, in 22 cases (48%) and beta-catenin accumulation in the nucleus in five cases (11%). Of four renal pelvis TCC examined, point mutation of exon 3 of the beta-catenin gene at codon 45 resulting in amino acid substitution (Ser to Phe) was detected in one (25%). The incidence of reduced expression of beta-catenin correlated significantly with the growth pattern (superficial type vs invasive type) of TCC (P < 0.05). These data indicate that: (1) aberrant beta-catenin expression may be at least partly involved in urothelial carcinogenesis, but less significantly so in renal carcinogenesis, and (2) it may be associated with the progression of TCC showing invasive growth.     

Berberine plays a cardioprotective role by inhibiting macrophage Wnt5a/β-catenin pathway in the myocardium of mice after myocardial infarction

Myocardial infarction (MI) is one of the diseases with high fatality rate. Berberine (BBR) is a monomer compound with various biological functions. And some studies have confirmed that BBR plays an important role in alleviating cardiomyocyte injury after MI. However, the specific mechanism is unclear. In this study, we induced a model of MI by ligation of the left anterior descending coronary artery and we surprisingly found that BBR significantly improved ventricular remodeling, with a minor inflammatory and oxidative stress injury, and stronger angiogenesis. Moreover, BBR inhibited the secretion of Wnt5a/β-catenin pathway in macrophages after MI, thus promoting the differentiation of macrophages into M2 type. In summary, BBR effectively improved cardiac function of mice after MI, and the potential protective mechanism was associated with the regulation of inflammatory responses and the inhibition of macrophage Wnt5a/β-catenin pathway in the infarcted heart tissues. Importantly, these findings supported BBR as an effective cardioprotective drug after MI.     

寒痹康汤对胶原诱导性关节炎大鼠Wnt/β-catenin和BMP/Smad信号通路的影响

目的:观察寒痹康汤对胶原诱导性关节炎(CIA)大鼠Wnt/β-连环蛋白(Wnt/β-catenin)和骨形态发生蛋白/Smad(BMP/Smad)信号通路的影响。方法:采用牛Ⅱ型胶原蛋白和弗氏不完全佐剂充分混合乳化制成CⅡ乳剂,进行皮下注射成功造模CIA大鼠模型,予灌服甲氨蝶呤作为对照,观察寒痹康汤高、中、低剂量提取物对CIA大鼠关节肿胀指数,以及Wnt/β-catenin和BMP/Smad信号通路相关蛋白表达的影响。结果:治疗4周后,各治疗组大鼠关节肿胀指数与模型对照组比较,差异有统计学意义(P<0.01)。寒痹康汤高剂量组与甲氨蝶呤组比较,差异有统计学意义(P<0.01);寒痹康汤中、低剂量组与甲氨蝶呤组比较,差异无统计学意义(P>0.05)。与空白对照组比较,模型对照组Wnt5α、Wnt7α蛋白表达升高,差异有统计学意义(P<0.05);与模型对照组比较,甲氨蝶呤组和寒痹康汤高、中、低剂量组Wnt5α、Wnt7α蛋白表达降低,差异有统计学意义(P<0.01);寒痹康汤高、中剂量组与甲氨蝶呤组Wnt5α、Wnt7α蛋白表达比较,差异无统计学意义(P>0.05)。与空白对照组比较,模型对照组β-catenin、BMP-2、BMP-4、BMP-5、Smad4、Smad5、Smad7蛋白表达升高,差异有统计学意义(P<0.05)。与模型对照组比较,甲氨蝶呤组和寒痹康汤高、中、低剂量组上述蛋白表达降低,差异有统计学意义(P<0.05);寒痹康汤高、中剂量组与甲氨蝶呤组上述蛋白表达比较,差异无统计学意义(P>0.05)。结论:寒痹康汤可以通过调节Wnt/β-catenin和BMP/Smad信号通路达到治疗类风湿关节炎及抗骨侵蚀的作用。     

冬凌草甲素抑制人卵巢癌细胞恶性行为及机制研究

目的 研究冬凌草甲素（Oridonin）对人卵巢癌（Human ovarian cancer）SKOV3 细胞迁移和侵袭能力的影响及其潜在的分子机制。方法 采用CCK-8 法检测不同浓度（0、5、10、20、40、80μmol/L）的冬凌草甲素作用SKOV3细胞24、48 和72h 后细胞活力的变化,计算半数抑制浓度（IC50）。采用Annexin V-FITC/PI 双染法检测SKOV3细胞凋亡。采用划痕修复实验检测SKOV3细胞的迁移能力,Transwell小室实验检测SKOV3的侵袭能力。Western blot检测SKOV3细胞上皮细胞间充质转化（EMT）蛋白［E-钙黏蛋白（E-cadherin）、波形蛋白（Vimentin）］和Wnt/β-连环蛋白（β-catenin）信号通路中β-catenin及下游靶分子原癌基因（C-myc）、细胞周期蛋白D1（Cyclin D1）蛋白的表达。结果 冬凌草甲素可抑制SKOV3 细胞活力,且具有时间-剂量依赖性,作用24、48 和72 h后IC50值为23.57、12.48和7.29μmol/L。与对照组比较,5、10和20μmol/L冬凌草甲素作用SKOV3细胞24h 后,细胞凋亡率明显升高,迁移率和侵袭率降低（P<0.05）。与对照组比较,5、10和20μmol/L冬凌草甲素可升高E-cadherin 蛋白表达（P<0.05）,降低Vimentin 表达（P<0.05）。与对照组比较,5、10和20 μmol/L冬凌草甲素可降低β-catenin、C-myc和Cyclin D1表达（P<0.05）。结论 冬凌草甲素具有抑制SKOV3细胞增殖、转移和侵袭能力的作用,该作用与其抑制Wnt/β-catenin 信号通路有关。