Identification of potential therapeutic target of naringenin in breast cancer stem cells inhibition by bioinformatics and in vitro studies

Cancer therapy is a strategic measure in inhibiting breast cancer stem cell (BCSC) pathways. Naringenin, a citrus flavonoid, was found to increase breast cancer cells’ sensitivity to chemotherapeutic agents. Bioinformatics study and 3D tumorsphere in vitro modeling in breast cancer (mammosphere) were used in this study, which aims to explore the potential therapeutic targets of naringenin (PTTNs) in inhibiting BCSCs. Bioinformatic analyses identified direct target proteins (DTPs), indirect target proteins (ITPs), naringenin-mediated proteins (NMPs), BCSC regulatory genes, and PTTNs. The PTTNs were further analyzed for gene ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, protein–protein interaction (PPI) networks, and hub protein selection. Mammospheres were cultured in serum-free media. The effects of naringenin were measured by MTT-based cytotoxicity, mammosphere forming potential (MFP), colony formation, scratch wound-healing assay, and flow cytometry-based cell cycle analyses and apoptosis assays. Gene expression analysis was performed using real-time quantitative polymerase chain reaction (q-RT PCR). Bioinformatics analysis revealed p53 and estrogen receptor alpha (ERα) as PTTNs, and KEGG pathway enrichment analysis revealed that TGF-ß and Wnt/ß-catenin pathways are regulated by PTTNs. Naringenin demonstrated cytotoxicity and inhibited mammosphere and colony formation, migration, and epithelial to mesenchymal transition in the mammosphere. The mRNA of tumor suppressors P53 and ERα were downregulated in the mammosphere, but were significantly upregulated upon naringenin treatment. By modulating the P53 and ERα mRNA, naringenin has the potential of inhibiting BCSCs. Further studies on the molecular mechanism and formulation of naringenin in BCSCs would be beneficial for its development as a BCSC-targeting drug.     

QuilliChew extended-release chewable tablets for the treatment of ADHD in patients ages 6 years old and above

ABSTRACT

Introduction: Attention-deficit/hyperactivity disorder (ADHD) is a common neurobehavioral disorder affecting as many as 6.4 million children and adolescents in the United States. Since amphetamine (AMPH) and methylphenidate (MPH) were found to be effective more than 60 years ago, numerous formulations of these compounds have been developed. New preparations have focused on convenience, with extended-release (ER) drugs allowing once-daily dosing. Multiple ER formulations do not require patients to swallow a tablet or capsule. Recent ER preparations include liquids, oral disintegrating tablets, and chewable tablets. Several new formulations use ion exchange technology containing both immediate-release and ER components.

Areas covered: Quillichew ER^TM (MPH-ERCT) is an ER methylphenidate designed to be chewed before swallowing. The technology and pharmacokinetics, along with efficacy and safety data, are presented.

Expert opinion: Extensive safety and efficacy data exist for MPH. ER formulations can be distinguished by preparation (tablet, capsule, liquid) and onset and duration of effect, but efficacy is similar for all ER MPH products. Each formulation has attributes, such as ease of titration, portability, and taste, that make it more acceptable for certain patients. Because AMPH and MPH are so effective, current technology research is focused on improving safety, convenience, and onset and duration of effect.     

乳结平胶囊对乳腺增生大鼠乳腺组织ER、PR表达的影响

目的:探讨乳结平胶囊对乳腺增生大鼠病理组织雌激素受体(ER)和孕酮受体(PR)表达的影响。方法:先采用苯甲酸雌二醇注射液连续im25d,后改用黄体酮注射液连续im5d,从而建立大鼠乳腺增生模型,以三苯氧胺为阳性对照药物,用乳结平胶囊高、低剂量治疗30d。结果:三苯氧胺组及乳结平胶囊高、低剂量组均能减轻大鼠乳头红肿或增生症状,降低大鼠血清雌二醇(E2)、催乳素(PRL)水平及升高孕酮(P)水平,降低子宫指数,明显减少导管上皮细胞层数和腺泡数,同时减少乳腺组织ER、PR的含量,抑制乳腺组织增生。结论:乳结平胶囊可通过调节雌激素水平,改善增生乳腺的病理形态变化,从而有效治疗大鼠乳腺增生。     

卵巢上皮性癌ER、PR和Ki67表达及其临床意义分析

[目的]研究ER、PR和Ki67在卵巢上皮性癌中的表达及其临床意义。[方法]用单克隆抗体、免疫组化技术测定74例原发性卵巢上皮性癌患者中ER、PR和Ki67的表达。[结果]ER、PR、Ki67的阳性表达率分别是59．5％、60．8％、74_3％，PR表达与临床分期相关。多因素分析显示PR表达、临床分期是独立的预后因素。[结论]在卵巢上皮性癌中，PR表达、临床分期具有独立的判断患者预后的价值。     

ER PR和p16联合检测在子宫内膜腺癌和宫颈腺癌鉴别诊断中的应用

目的:探讨ER,PR和p16联合免疫组化染色在子宫内膜腺癌和宫颈腺癌鉴别诊断中的价值.方法:采用免疫组化SP法染色,观察ER,PR,p16在子宫内膜腺癌和宫颈腺癌中的表达情况.结果:子宫内膜腺癌中ER,PR和p16的阳性率分别为85.29%、76.47%、23.52%;宫颈腺癌为33.33%、42.86%、80.95%,子宫内膜腺癌ER,PR的阳性率明显高于宫颈腺癌,宫颈腺癌p16的阳性率明显高于子宫内膜腺癌.在子宫内膜腺癌和宫颈腺癌中3种标记差异均有显著性(ER:x2=20.78 P＜0.005;PR:x2=8.32 P＜0.01;p16 x2=17.24 P＜0.005).结论:ER,PR,p16联合检测对鉴别子宫内膜腺癌和宫颈腺癌有一定实用价值.     

PTEN与预后相关因子在乳腺癌组织芯片中的相关性

目的:构建乳腺癌组织芯片,研究PTEN与乳腺癌预后相关因子ER、PR、c-erbB-2、Ki67在原发性乳腺癌中的蛋白表达及其相关性。方法:制作乳腺癌组织芯片,用免疫组织化学法检测芯片上165例乳腺癌组织中PTEN、ER、PR、c-erb B-2和Ki67蛋白的表达。结果:乳腺癌组织中PTEN的高表达例数为66/163(40.5%),ER、PR、c-erb B-2和Ki67阳性率分别为51.5%、35.6%、34.4%、44.8%。PTEN阳性表达随腋淋巴结转移数、组织学分级和TNM分期的增高而降低,呈负相关性P<0.05),与肿瘤的大小和发生年龄无明显相关。乳腺癌中PTEN与ER、PR蛋白表达呈显著正相关,与c-erb B-2、Ki67蛋白表达呈负相关。结论:PTEN蛋白低表达及表达缺失与性激素失调及乳腺肿瘤细胞的生长、预后之间存在一定的联系,PTEN可作为判断乳腺癌恶性程度的潜在标志物。同时检测PTEN、ER、PR、c-erb B-2和Ki67对乳腺癌的辅助治疗具有重要意义及实用价值。     

大豆异黄酮和钙对绝经妇女骨代谢的影响及与ER基因Px单倍型的关系

目的探讨补充大豆异黄酮和钙对绝经妇女骨代谢的影响及与ER基因Px单倍型的关系。方法应用多聚酶链反应-限制性片断长度多态性(PCR-RFLP)检测691名汉族45～65岁绝经妇女的ER-α的PvuII和XbaI酶切位点多态性,从497名PvuII-XbaI单倍体确定者中随机抽取93人测定BMD,将T<-1.5者纳为干预对象(共60人)进行为期一年的随机对照干预实验。将干预对象随机分为两组:补钙+VD组和补钙+VD+大豆异黄酮组。给予的剂量分别为:钙440mg/d;VD100IU/d;大豆异黄酮100mg/d,干预期为12mon。测定基线和终期的各项骨代谢生化指标。结果干预12个月后,补钙组和补钙+大豆异黄酮组,机体的雌激素水平均未见显著性的改变,而25-OH-D均水平升高(P=0.09),但两组间的变化率无显著性的差异;血清钙磷及反映骨形成的指标未见显著性改变。补钙组的抗酒石酸酸性磷酸酶-5b(Tracp-5b)较基线升高[变化率:23.4%(-22.9%～49.1%)],而补钙+大豆异黄酮组则较基线降低[变化率:-4.2%(-26.0%～17.6%)],两组的变化率有显著性差异(P<0.05),尿吡啶交联物(Pyd)和I型胶原N末端肽(NTx)则未见显著性改变。补钙组,non-Px基因型较Px基因型更容易出现Pyd升高,骨特异性碱性磷酸酶(BAP)降低。钙+大豆异黄酮的干预效果在两种基因型间没有差异.结论补充大豆异黄酮并不影响体内的雌激素水平。反应骨吸收的生化指标Tracp-5b对大豆异黄酮的作用较敏感。单纯补钙后骨代谢生化指标的改变受Px单倍体基因型的影响。     

The mechanism of BRCA1 participate sporadic breast carcinomas genesis

Objective To elucidate the BRCA1 participated mechanism of genesis and development of sporadic breast cancer through detect the statues of BRCA1 and analysis the relationship with the pathologic and clinic parameters.Methods BRCA1 statues were respectively analyzed in frozen samples or paraffine fixed sporadic breast carcinoma and benign breast tissues by three methods:protein expression by immunohistochemistry(IHC),the methylation of BRCA1 promoter by methylation specific PCR(MSP),gene copy number by interphase fluorescence in situ hybridization(FISH).Results 14.2%(29/204)cases were detected hypermethylation of BRCA1 promoter in sporadic breast cancer.BRCA1 mean copy number in sporadic breast cancer(1.70±0.14)less than those in benign tissues(2.03±0.08,P<0.05),and in sporadic breast cancer with hypermethylation of BRCA1(1.62±0.09)significantly less than in those without hypermethylation(1.84±0.26,P<0.05).The loss copy related to the methylation of BRCA1 promoter.There were significant of 41.1%(88/214)cases no BRCA1 nuclei expression in sporadic breast cancers.Loss expression of BRCA1 had significant correlation with higher histological stages,axillary's lymph nodal metastasis(P<0.01),lower expression of ERα,and overexpression of HER-2 protein(P<0.01).Conclusions There are BRCA1 methylations,loss BRCA1 gene copy and loss protein expression in the sporadic breast cancer,the three statues of BRCA1 is correlated to each other;and the loss expression of BRCA1 protein related to part of pathology and clinic parameters.     

补肾调经系列方对功能失调性子宫出血患者生殖激素及子宫内膜激素受体的影响

目的探讨补肾调经系列方对功能失调性子宫出血患者生殖激素及子宫内膜雌激素受体(ER)、孕激素受体(PR)的影响。方法运用补肾调经系列方周期给药对39例功能失调性子宫出血患者进行治疗,采用磁分离酶免激素定量测定法测定患者治疗前后血浆雌二醇(E2)、孕酮(P)、促卵泡素(FSH)、黄体生成素(LH);采用流式细胞术测定子宫内膜ER、PR。结果与治疗前相比,治疗后患者子宫内膜ER表达量明显降低(P<0.05),E2水平明显升高(P<0.01)。结论补肾调经系列方有调节生殖激素和子宫内膜ER的作用。     

甲状腺癌中ER的表达及相关性研究

目的 检测甲状腺病变中雌激素受体（ER）的表达,探讨其在甲状腺癌的临床病理的生物学性意义.方法 应用免疫组化SP法检测ER在128例甲状腺良、恶性组织的表达情况.结果 甲状腺癌中ER阳性表达率明显高于甲状腺良性病变中ER的表达,且与甲状腺癌的组织分化程度、淋巴结转移相关,但与患者性别、年龄无相关性.结论 ER的检测可作为反映甲状腺病变良、恶性的一个生物学指标.