Xanthatin,a novel potent inhibitor of VEGFR2 signaling,inhibits angiogenesis and tumor growth in breast cancer cells

Anti-angiogenesis targeting vascular endothelial growth factor receptor 2 (VEGFR2) has emerged as an important tool for cancer treatment. In this study, we described a novel VEGFR2 inhibitor, xanthatin, which inhibits tumor angiogenesis and growth. The biochemical profiles of xanthatin were investigated using kinase assay, migration assay, tube formation, Matrigel plug assay, western blot, immunofluorescence and human tumor xenograft model. Xanthatin significantly inhibited growth, migration and tube formation of human umbilical vascular endothelial cell as well as inhibited vascular endothelial growth factor (VEGF)-stimulated angiogenesis. In addition, it inhibited VEGF-induced phosphorylation of VEGFR2 and its downstream signaling regulator. Moreover, xanthatin directly inhibit proliferation of breast cancer cells MDA-MB-231. Oral administration of xanthatin could markedly inhibit human tumor xenograft growth and decreased microvessel densities (MVD) in tumor sections. Taken together, these preclinical evaluations suggest that xanthatin inhibits angiogenesis and may be a promising anticancer drug candidate.     

三阴乳腺癌的治疗新进展

三阴乳腺癌是乳腺癌的一种特殊亚型,侵袭性强、预后差,除化疗外几乎没有其他的治疗措施.标准的辅助化疗和新辅助化疗应用蒽环类、环磷酰胺和紫杉醇类药物.许多研究报道了三阴乳腺癌的分子学特性,由此,我们可针对一些靶点来进行治疗.对铂类药物治疗三阴乳腺癌,尤其是BRCA基因突变的三阴乳腺癌的实验数据分析发现铂类药物在治疗中占主导作用.除此之外,我们回顾近期的药物试验数据,讨论聚ADP核糖聚合酶抑制剂、雄激素受体抑制剂等的作用.     

Expression of metabolism-related proteins in triple-negative breast cancer

To investigate the dominant metabolic type of triple-negative breast cancer (TNBC) and evaluate its clinical implication through analysis of protein expression related to glycolysis, glutaminolysis, and mitochondrial oxidative phosphorylation. Tissue samples from 129 patients with TNBC who underwent mastectomy due to invasive breast cancer from 2000 to 2005 were prepared for tissue microarray. By immunohistochemical staining of the tissue microarrays, the markers of glycolysis-related proteins (Glut-1, CAIX, MCT4), glutaminolysis-related proteins (GLS1, GDH, ASCT2), and mitochondrial enzymes (ATP synthase, SDHA and SDHB) were analyzed. Based on the results, the metabolic phenotypes were defined based on positivity for more than two of three markers for each phenotype as follows: glycolysis type (Glut-1, CAIX and MCT4), glutaminolysis type (GLS1, GDH and ASCT2) and mitochondrial type (ATP synthase, SDHA and SDHB). The percentages of samples with metabolic phenotypes of tumor and stroma of TNBC were as follows: for tumor, mitochondrial type (85.3%) > glutaminolysis type (67.4%) > glycolysis type (63.0%); and for stroma, glutaminolysis type (37.2%) > glycolysis type (16.3%) > mitochondrial type (14.0%). The most common metabolic phenotype of TNBC was glycolysis type for basal-like type and non-glycolysis type for non-basal-like type (p=0.047). The correlation between glutaminolysis and mitochondrial type was statistically significant in both tumor and stroma (p<0.001). In conclusion, tumor cells of TNBC express glycolysis and mitochondrial metabolism-related proteins. Glycolysis type is the most common phenotype of basal-like type, and reversely, non-glycolysis type is the most common phenotype of non basal-like type.     

siRNA抑制ErbB2基因的表达及对乳腺癌细胞生长的影响

目的:设计并构建ErbB2的小干扰RNA,检测其对ErbB2蛋白表达的干扰效果,并检测其对乳腺癌细胞ZR75-1生长的影响。方法:设计2条针对ErbB2的siRNA,并克隆到siRNA表达载体pSliencer 2.1-U6 neo上。经酶切和测序证明构建成功后,将其与pcDNA3-FLAG-ErbB2共转染于293T细胞,以及单转siRNA于乳腺癌SKBR3和ZR75-1细胞,通过Western blot分别检测siRNA对外源和内源ErbB2的干扰效果。通过结晶紫实验研究siRNA对ZR75-1生长的影响。结果:Western blot证明构建的两条ErbB2 siRNA均能有效抑制外源和内源ErbB2蛋白的表达,并抑制乳腺癌ZR75-1细胞的生长。结论:构建的siRNA能有效地抑制ErbB2蛋白的表达并抑制ZR75-1细胞的生长。     

靶向TAM在三阴性乳腺癌治疗中的研究进展

三阴性乳腺癌(triple negative breast cancer,TNBC)是乳腺癌中预后最差的类型,严重威胁女性生命健康。化疗和免疫治疗是TNBC最常用的治疗手段。目前临床使用抗程序性死亡受体1(programmed cell death protein 1,PD-1)和抗程序性死亡配体1(programme...     

三阴性乳腺癌患者BRCA1和BRCA2基因突变的临床特征及预后因素

目的:探究三阴性乳腺癌患者BRCA1和BRCA2基因突变检测的临床意义及预后因素分析.方法:研究对象选取为2003年1月至2015年12月之间的三阴性乳腺癌156例,所有患者均通过PCR法和DNA序列测定检验BRCA1和BRCA2基因突变情况,分析乳腺癌患者BRCA1和BRCA2基因突变特点.应用Log-Rank检验对BRCA1和/或BRCA2基因突变的三阴性乳腺癌患者的各项指标如年龄、ECOG状态、临床分期、淋巴结阳性数、月经状态和给药方式进行单因素分析.应用Cox风险比例回归模型分析患者年龄、ECOG状态、临床分期、淋巴结阳性数、肿瘤大小、月经状态和给药方式等多因素分析.结果:三阴乳腺癌患者发生基因突变21例,总体发生率13.46％,BRCA1突变15例,BRCA2突变6例.Log-Rank检验结果显示,发病年龄越大,ECOG评分越高,临床分期越晚,淋巴结阳性数越多,预后越差(P＜0.05),而发病时月经状态和给药方式与预后无关.COX风险比例回归模型显示,肿瘤大小(相对危险度,3.163;95％CI:1.455～9.287;P＜0.05)和淋巴结转移数(相对危险度,1.859;95％CI:1.254～6.875;P＜0.05)是BRCA基因突变三阴性乳腺癌独立的预后因素.结论:BRCA1和BRCA2基因突变可能与乳腺癌尤其是三阴乳腺癌可能有着密切的相关性,发病年龄、ECOG评分,临床分期和淋巴结阳性数及肿瘤大小与BRCA基因突变的三阴乳腺癌预后有关.     

三阴型乳腺癌中NQO1蛋白过表达的临床病理意义

目的探讨醌氧化还原酶[NAD(P)H:quinone oxidoreductase-1,NQO1]在三阴型乳腺癌(triple-negative breast cancer,TNBC)中过表达的临床病理意义。方法应用免疫荧光染色法观察NQO1蛋白在MDA-MB-468细胞中的表达定位,并应用免疫组化SP两步法检测NQO1蛋白在87例TNBC组织、45例导管内原位癌(ductal carcinoma in situ,DCIS)组织以及36例癌旁非瘤乳腺上皮组织中的表达,同时分析其过表达与TNBC临床病理特征间的关系。结果 NQO1蛋白主要表达于TNBC细胞胞质中。在TNBC组织中,NQO1蛋白的阳性率和强阳性率分别为75.9%(66/87)和62.1%(54/87),显著高于DCIS组织和癌旁非瘤乳腺上皮组织(P均0.01)。NQO1蛋白表达水平与TNBC患者的临床分期和淋巴结转移情况密切相关(P均0.05),但与患者年龄、绝经情况、肿瘤大小及组织分化程度无关(P均0.05)。结论 NQO1蛋白在TNBC组织中明显高表达,其表达水平与TNBC患者预后密切相关,有望成为TNBC治疗的新分子靶标。     

青蒿琥酯对吉西他滨体外抗胰腺癌活性的作用

目的:探讨青蒿琥酯辅助治疗对吉西他滨抗胰腺癌活性的影响和机制。方法:分别采用分子克隆及RNA干扰方法于人胰腺癌细胞Capan-2中过表达和干扰鼠双微体蛋白2(MDM2),MTT实验检测p53野生型胰腺癌细胞系Capan-2的细胞活力。Western blot实验检测Capan-2细胞中MDM2、p53、Noxa和Puma的表达水平,细胞色素C和凋亡诱导因子的释放,以及caspase-9和caspase-3的活化。流式细胞术检测Capan-2细胞的线粒体膜电位和凋亡水平。结果:吉西他滨联合青蒿琥酯组Capan-2的相对细胞活力显著低于吉西他滨单处理组(P 0. 05)。青蒿琥酯处理显著抑制Capan-2细胞中MDM2的表达水平(P 0. 05)。吉西他滨联合青蒿琥酯组的p53、Noxa及Puma表达水平均明显高于吉西他滨单处理组(P 0. 05)。青蒿琥酯明显促进吉西他滨依赖的Capan-2细胞线粒体膜电位的下降,细胞色素C和凋亡诱导因子的释放,caspase-9和caspase-3的活化,以及凋亡的发生。转染MDM2表达质粒后,青蒿琥酯联合吉西他滨对Capan-2细胞的凋亡诱导途径受到显著抑制。结论:青蒿琥酯通过MDM2/p53途径提高吉西他滨的抗胰腺癌活性。     

The effect of lipocisplatin on cisplatin efficacy and nephrotoxicity in malignant breast cancer treatment

A lipid–cisplatin conjugate was synthesized for super-molecular assembly with lipids to form a new generation of liposomal cisplatin formulation, lipocisplatin. In vitro, lipocisplatin has higher efficacy in human ovarian cancer A2780 and human breast cancer MCF-7 with the murine breast cancer cell line 4T1 which is currently an established model for stage IV breast cancer as the most sensitive strain. Moreover, lipocisplatin demonstrated a greater MTD value and relatively longer blood circulation as compared to cisplatin. Lipocisplatin preferentially accumulate drugs to the tumor site, resulting in a better tumor inhibition efficacy. Moreover, lipocisplatin exceeds the size cutoff for kidney clearance, hence it bypasses the nephrotoxicity of cisplatin which is a major curse of one of the most efficient anticancer drugs nowadays in clinic. The results here indicated lipocisplatin may be translated into a new generation of liposomal based cisplatin drug in clinic.     

Expression of VEGF and VEGFR2 in tumors during neoadjuvant therapy of patients with breast cancer

We revealed a significant positive correlation between the concentrations of tissue VEGF and its receptor VEGFR2 in the primary tumor. The degree and direction of changes in the content of VEGF and VEGFR2 in cytosols of tumors from patients with locally spread breast cancer during the preoperation therapy depended on initial levels of these parameters. We found that the expression of the studied factors depended on the degree of therapeutic pathomorphosis in the tumor tissue. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 2, pp. 206–209, February, 2008     

COX-2抑制剂在乳腺癌药物治疗中的研究进展

环氧化酶-2(COX-2)在乳腺癌细胞中过度表达,影响乳腺癌的发生、发展和预后,可作为预防和治疗的靶分子。选择性COX-2抑制剂可联合化疗、内分泌治疗及生物治疗等,作为肿瘤治疗的辅助药物应用于临床,为乳腺癌的治疗提供一种新的途径。     

Aldehyde dehydrogenase 1 expression predicts poor prognosis in triple-negative breast cancer

Ohi Y, Umekita Y, Yoshioka T, Souda M, Rai Y, Sagara Y, Sagara Y, Sagara Y & Tanimoto A
(2011) Histopathology 59 , 776–780 Aldehyde dehydrogenase 1 expression predicts poor prognosis in triple‐negative breast cancer Aims: Aldehyde dehydrogenase 1 (ALDH1) has been identified as a reliable marker of breast cancer stem cells, and its clinical significance as a prognostic indicator of breast cancer has been reported by several investigators. However, the clinical significance of ALDH1 expression in triple‐negative (TN) breast cancer, a high‐risk breast cancer lacking the benefit of specific therapy, remains to be solved. Methods and results: We performed immunohistochemical analyses of 106 TN breast cancers, using paraffin‐embedded sections. The basal‐like phenotype was also investigated with the use of basal cytokeratin 5/6 and epidermal growth factor receptor. ALDH1 expression in carcinoma cells was found in 59% of cases and was correlated with high histological grade alone (P < 0.006), whereas ALDH1 expression in stromal cells was found in 49% of cases but was not correlated with any clinicopathological parameter. Patients with ALDH1 expression in carcinoma cells had a shorter relapse‐free survival (RFS) according to the log‐rank test (P = 0.015). According to Cox multivariate analysis, ALDH1 expression in carcinoma cells was an independent prognostic indicator of RFS (P = 0.025). The log‐rank test revealed that stromal expression of ALDH1 had no effect on RFS. Conclusions: ALDH1 expression in carcinoma cells is an independent prognostic factor in TN breast cancer patients.     

Down-regulation of BRMS1 by DNA hypermethylation and its association with metastatic progression in triple-negative breast cancer

Breast cancer metastasis suppressor 1 (BRMS1) is a metastasis suppressor gene in several solid tumors. However, the expression and function of BRMS1 in triple-negative breast cancer (TNBC) have not been reported. In this study, we found that BRMS1 was down-regulation in breast cancer cell lines and primary TNBC, while decreased expression of BRMS1 mRNA was significantly associated with lymph node metastasis. And this down-regulation was found to be in accordance with aberrant methylation of the gene. Hypermethylation of the gene was observed in 53.4% (62/116) of the TNBC primary breast carcinomas, while it was found in only 24.1% (28/116) of the corresponding nonmalignant tissues. In addition, BRMS1 expression was restored in MDA-MB-231 after treatment with the demethylating agent, 5-aza-2-deoxycytidine (5-Aza-dC), and demethylation of the highly metastatic cells MDA-MB-231 induced invasion suppression of the cells. Furthermore, the suppression of BRMS1 by siRNA transfection enhanced cancer cells invasion. Collectively, our results suggest that the aberrant methylation of BRMS1 frequently occurs in the down-regulation of BRMS1 in TNBC and that it may play a role in the metastasis of breast cancer.     

Alterations of microRNAs are associated with impaired growth of MCF-7 breast cancer cells induced by inhibition of casein kinase 2

Background and aim: Protein Kinase (casein kinase 2, CK2) is a pleiotropic serine-threonine kinase that is frequently dysregulated in many human tumors; microRNAs (miRNAs) are a class of small noncoding RNAs which play important roles in human cancers. This study aimed to investigate the role of CK2 and miRNA expression in breast cancer. Methods: Casein kinase 2 in MCF-7 breast cancer cell line was inhibited by the CK2 inhibitor TBB (4,5,6,7-tetrabromobenzotriazole), then cell proliferation was studied using MTT assay, and cell cycle distribution and apoptosis were detected by flow cytometry. The alteration of microRNAs expression profile was determined by microarray technology, followed by RT-PCR confirmation. Results: Here, we for the first time showed that inhibition of CK2 in MCF-7 breast cancer cells causes suppressed cell growth, which was related with dysregulation of the miRNA profile and altered expression. CK2 inhibition induced the up-regulated expression of 17 miRNAs and 10 down-regulated microRNAs which contributed to the impaired growth, inhibited cell cycle progress and increased apoptosis of MCF-7 cells by a CK2 inhibitor. Conclusions: These findings highlight the potential role of dysregulated miRNA expression regulated by CK2 in breast cancer.     

Oridonin exhibits anti-angiogenic activity in human umbilical vein endothelial cells by inhibiting VEGF-induced VEGFR-2 signaling pathway

Oridonin has been found to be a potential anti-angiogenesis agent. However, its functional targets and the underlying mechanisms are still vague. In vitro studies we found that oridonin not only inhibited VEGF-induced cell proliferation, migration and tube formation but also caused G2/M phase arrest and triggered cellular apoptosis in HUVECs. In mechanistic studies revealed that oridonin exhibited the anti-angiogenic potency, at least in part, through the down-regulation of VEGFR2-mediated FAK/MMPs, mTOR/PI3K/Akt and ERK/p38 signaling pathways which led to reduced invasion, migration, and tube formation in HUVECs. Our results could provide evidence that oridonin exerts strong anti-angiogenesis activities via specifically targeting VEGFR2 and its signaling pathway.