薯蓣皂苷元通过NF-κB信号通路抑制三阴性乳腺癌的机制研究

目的观察薯蓣皂苷元对乳腺癌细胞增殖与迁移的抑制作用,探讨其作用机制与NF-κB信号通路的关系。方法采用CCK-8法检测不同浓度的薯蓣皂苷元作用乳腺癌细胞株及正常细胞株24 h后的生长抑制率。倒置相差显微镜下观察薯蓣皂苷元作用MDA-MB-231细胞24 h后,细胞形态的变化。细胞划痕法观察薯蓣皂苷元对MDA-MB-231细胞迁移的影响。RT-PCR检测凋亡相关基因Bcl-2 mRNA表达情况。Western Blot法检测薯蓣皂苷元对MDA-MB-231细胞凋亡相关蛋白Bax、Bcl-2的影响,以及对NF-κB信号通路相关蛋白P65的影响。结果不同浓度薯蓣皂苷元对两种人类三阴性乳腺癌细胞株和小鼠E0771细胞均有显著的抑制效应,但对End1/E6E7细胞影响较小,只有在极高浓度作用下才产生抑制效应。薯蓣皂苷元作用MDA-MB-231细胞后,细胞的形态发生改变,细胞边缘回缩,失去正常上皮生长样形态。薯蓣皂苷元可显著抑制MDA-MB-231细胞的迁移,并呈浓度和时间依赖性。薯蓣皂苷元作用MDA-MB-231细胞24 h后,Bcl-2 mRNA表达量低于对照组。Bcl-2、P65蛋白表达量低于空白组,Bax蛋白表达量高于对照组。结论薯蓣皂苷元对乳腺癌细胞的抑制作用明显,其作用机制可能与抑制NF-κB信号通路的激活有关。

PAK4在乳腺癌及良性乳腺病变中的表达及意义

目的探讨p21活化激酶4(p21-activatedkinase-4,PAK4)在乳腺癌发病、发展及转移过程中的作用。方法采用免疫组化SP法检测35例正常乳腺组织、22例乳腺囊性增生病、28例乳房纤维腺瘤、37例乳腺癌(其中非浸润性癌7例,早期浸润癌9例,浸润癌21例)及13例乳腺癌转移瘤组织中的PAK4含量,比较各类组织中PAK4的表达特点与分布特征。结果 PAK4表达主要位于胞浆,胞核偶有少量表达,而乳腺腺体细胞外基质中基本不表达;正常乳腺组织、良性病变(乳腺囊性增生病和乳腺腺瘤)、乳腺癌和乳腺癌转移瘤组织中的PAK4表达总体阳性率依次升高;乳腺非浸润性癌、早期浸润癌、浸润癌组织中PAK4的表达阳性率亦呈依次升高趋势。结论 PAK4与乳腺癌的演进和转移密切相关,PAK4及其基因有可能成为乳腺癌诊断和治疗的新靶点。

四君子汤对人乳腺癌MDA-MB-231细胞生长及PD-L1表达的影响

目的:观察四君子汤对三阴性乳腺癌(triple negative breast cancer, TNBC)细胞MDA-MB-231增殖、凋亡及程序性死亡分子配体-1(programmed cell death ligand 1, PD-L1)表达的影响。方法:以四君子汤煎煮浓缩液处理人TNBC细胞MDA-MB-231,分别在24,48,72 h,采用细胞增殖/毒性实验的方法检测高(2 g/L)、中(1 g/L)、低剂量(0.1 g/L)四君子汤在不同时间点对MDA-MB-231细胞增殖的影响;采用流式细胞术检测高、中、低剂量四君子汤在48 h时对MDA-MB-231细胞凋亡的影响;采用Western blot技术检测加药48 h时四君子汤高、中、低剂量组MDA-MB-231细胞中PD-L1的表达情况。结果:与空白对照组对比,3种剂量四君子汤处理不同时间对细胞增殖影响无显著性差异;四君子汤处理72 h后,高、中、低剂量组抑制率分别为10.21%、2.97%和0.12%,其中高剂量组抑制率显著低于紫杉醇组(49.88%),差别有统计学意义(P<0.05)。四君子汤高、中、低剂量组48 h,凋亡率分别是24.25%、16.90%和15.37%,与空白对照组对比,四君子汤高剂量组凋亡率显著增高(P<0.05)。与空白对照组对比,四君子汤高剂量组处理48h后,PD-L1表达显著降低(P<0.05)。结论:四君子汤对TNBC细胞MDA-MB-231的增殖无明显影响,高剂量四君子汤能够诱导其凋亡,并降低PD-L1的表达。

类器官在乳腺癌中的研究进展

乳腺癌已成为中国妇女发病率最高的肿瘤,其发病率呈逐年增高趋势,为女性健康的重大隐患。因此,实现乳腺癌患者的个体化治疗尤为重要。乳腺癌类器官是一种利用患者癌组织干细胞进行体外培养的三维模型,能够保持原发肿瘤异质性和肿瘤生长的微环境,在疾病建模、肿瘤发生机制和药敏试验等方面提供了可靠的模型。本文概述了乳腺癌类器官的基础、机制研究、药敏试验及新药研发等方面,为乳腺癌的个体化治疗提供新的方向。

乳腺癌患者外周血中趋化因子及其受体表达水平变化在乳腺癌发病机制中的作用

目的探讨乳腺癌患者外周血中趋化因子及其受体表达水平变化在乳腺癌发病机制中的作用,为临床诊治奠定理论基础。方法选择2016年4月-2019年5月来嘉兴市第一医院治疗的乳腺癌患者108例(观察组)和同期来该院做体检的108例(对照组)妇女作为研究对象。检查后对患者的病症进行评估,比较不同分子分型乳腺癌患者的临床资料和两组患者血清趋化因子含量水平、外周血CD3+T细胞差异。结果不同分子分型乳腺癌患者的年龄、肿瘤发生部位、所在象限、肿瘤直径、病理分型、淋巴转移情况、月经状态、临床分期和术后化疗等临床特征比较,差异有统计学意义(均P<0.05)。观察组患者的血清趋化因子CCL20、RANTES、MCP-1、IL-8含量水平均高于对照组,差异有统计学意义(均P<0.05)。两组患者的外周血CD3+T细胞差异无统计学意义(P>0.05)。观察组患者的CXCR1和CCR2均低于对照组,CCR6和CCR5均高于对照组,差异有统计学意义(均P<0.05)。结论将外周血中趋化因子及其受体表达水平变化用于探讨乳腺癌患者的预测指标效果显著,CCL20、RANTES、MCP-1、IL-8各项指标的升高能够为预测乳腺癌患者的发病及病程预测起到重要作用,对外周血CD3+T细胞进行分析,对预后治疗效果更佳,具有很高的临床应用价值。

The Immuno-fluorescence Quantity Analysis of α-Tubulin and γ-Tubulin Protein in Precancerous Lesion and Carcinoma of the Breast and Its Significance

OBJECTIVE To investigate the changes and values of the expression of α-tubulin and γ-tubulin in atypical ductal hyperplasia （ADH）, ductal carcinoma in situ （DCIS） and invasive ductal carcinoma （IDC） of the breast. The relationship between centrosome abnormalities and breast tumor development was further discussed. METHODS There were three groups including ADH, DCIS and IDC with 30 cases in each group. They were analyzed by immuno-fiuorescence quantity analysis. The expression levels of α-tubulin and γ-tubulin protein in these tissues were detected by flow cytometry immuno-fiuorescence analysis and compared with the results from normal tissues. Immunohistochemistry was also performed in this research. RESULTS The results showed significant differences of the average of the positive （FITC labeled） cells （P=0.000） among the four groups. The level of the IDC group was the highest, while normal breast tissue showed the lowest level. The results suggested that the expression levels of α-tubulin and γ-tubulin both increased as the grade of cellular proliferation and differentiation increased. The expressions showed significant differences among all the groups, except between the ADH and DCIS. There were no significant differences between α-tubulin and γ-tubulin expression in each group （P〈0.05）, as there was agreement in the immuno-fluorescence and immunohistochemical analysis for protein expression. CONCLUSION There is abnormal expression of centrosome tubulin as an early event in the development of breast tumor. Furthermore these aberrations may play a key role during oncogenesis and promote cellular transformation to malignancy. The immuno-fiuorescence quantitive analysis and immunohistochemistry can complement each other.

Replication-selective Oncolytic Adenovirus CNHK300 in the Treatment of Breast Cancer Cell Lines in vitro

Objective： To evaluate the tumor selectivity and therapeutic efficiency of replication-competent adenovirus CNHK300 on human breast cancer cells. Methods： RT-PCR was used to detect the hTERT mRNA activity in various breast cancer and normal fibroblast cell lines. Virus proliferation assay, cell viability assay and Western blot were applied to evaluate the proliferation and cytolysis selectivity of CNHK300. Results： The telomerase activity of MCF-7, BT-549 and SK-BR-3 was positive, while telomerase in MRC-5 and BJ was negative. The progeny virus titers in MCF-7, BT-549 and SK-BR-3 after 48 h of CNHK300 exposure was 40 625, 1 265 and 20 000 fold higher than those of 0 h, even slightly higher than those of wtAd5 （except in SK-BR-3）. ONYX-015 virus proliferation ability was weaker than that of CNHK300 in cancer cells. However, CNHK300 exhibited attenuated replicative ability as compared with wtAd5 in MRC-5 and BJ. The CNHK300 replicatative multiple was 63 and 192 fold at 48 h respectively, while the wtAd5 still multiplied 3 160-4 846 fold. CNHK300 could cause about half of breast cancer cells to die within 7 days at MOI 10 pfu/cell and below, whereas the IC50 in BJ and MRC-5 was as high as MOI 100 pfu/cell. CNHK300 E1A protein could be detected in breast cancer cells and 293 cells but not in normal fibroblast cells. Conclusion： hTERT promoter can successfully modulate the CNHK300 to be selectively replicated in breast cancer cells positive for telomerase, which may be a potential treatment strategy in breast cancer.

Metastatic breast cancer to the gastrointestinal tract:A case series and review of the literature

Metastatic breast cancer involving the hepatobiliary tract or ascites secondary to peritoneal carcinomatosis has been well described. Luminal gastrointestinal tract involvement is less common and recognition of the range of possible presentations is important for early and accurate diagnosis and treatment. We report 6 patients with a variety of presentations of metastatic breast cancer of the luminal gastrointestinal tract. These include oropharyngeal and esophageal involvement presenting as dysphagia with one case of pseudoachalasia, a linitis plastica-like picture with gastric narrowing and thickened folds, small bowel obstruction and multiple strictures mimicking Crohn’s disease, and a colonic neoplasm presenting with obstruction. Lobular carcinoma, representing only 10% of breast cancers is more likely to metastasize to the gastrointestinal tract. These patients presented with gastrointestinal manifestations after an average of 9.5 years and as long as 20 years from initial diagnosis of breast cancer. Given the increased survival of breast cancer patients with current therapeutic regimes, more unusual presentations of metastatic disease, including involvement of the gastrointestinal tract can be anticipated.

Activation of NF-κB in Human Breast Cancer and its Role in Cell Proliferation and Progresssion

OBJECTIVE To investigate the expression of the nuclear transcription factor NF-κB, ER, HER2 and PCNA in breast cancers, and to study the relationship between activation of NF-κB and clinicopathologic parameters including the level of PCNA, ER, HER2, lymph node involvement, tumor size and histological grade （differentiation）. METHODS Sixty cases of human breast cancer tissues and adjacent non-neoplastic breast tissues were examined for NF-κB, HER2 and ER, as well as PCNA by immunohistochemical methodS. In addition the clinicopathologic parameters of the patients including lymph node involvement, tumor size and histological grade （differentiation） were collected. RESULTS The expression of NF-κB in the breast cancers and adjacent non-neoplastic breast tissue was 50.0% （30/60） and 40.0% （24/60） respectively, resulting in no significant difference （P〉0.05）. NF-κB and HER2 expression was positively correlated whereas NF-κB and ER expression was negatively correlated. The NF-κB activation was 77.8% （14/18） in the breast cancers that were ER-/HER2^＋, a level significantly higher （P〈0.001） in comparison to the other groups of patients. The expression of NF-κB in the low-differentiated group （grade Ⅲ） was 57.1%, and in the moderate-differentiated group （grade Ⅱ） was 50.3%, both of which were higher than the 35.7% found in the high-differentiated group （grade Ⅰ）. NF-κB activation in the cancers was significantly correlated with the histological grade （P〈0.05）, PCNA expression （P=0.003） and lymph node involvement and tumor size （P=0.03 and 0.002, respectively）. CONCLUSION NF-κB was activated abnormally in a portion of the breast cancers. The finding that NF-κB activation was positively correlated with HER2 expression, the level of PCNA, tumor grade, size and lymph node involvement is in accord with the ability of NF-κB to promote cellular proliferation and migration, clearly identifies the protein as a hallmark for targeted dysregulation in oncogenesis. NF-κB may be a hopeful target for breast cancer therapy.

Gastric signet-ring adenocarcinoma presenting with breast metastasis

Breast metastases from gastric cancer are extremely rare. A case report of a 37-year-old female with breast inflammatory invasion and ascites is described. Breast biopsy revealed carcinomatous invasion of the lymphatics from adenocarcinoma cells with signet-ring features. Estrogen （ER） and progesterone receptors （PR） and c-erb-B2 were negative. Upper gastrointestinal endoscopy revealed a prepyloric ulcerative mass. Histopathologic examination of the lesion showed infiltration from a highgrade adenocarcinoma, identical with that of the breast. Immunostaining was positive for cytokeratins CK-7 and CK-20 and CEA and negative for ER and PR. Ascitic fluid cytology was positive for adenocarcinoma cells. Mammography was not diagnostic. Abdominal CT scanning revealed large ovarian masses suggestive of metastases （Krukenberg＇s tumor）. A cisplatin-based regimen was given but no objective response was observed. The patient died six months after initial diagnosis. A review of the literature is performed.

Effect of Neoadjuvant CAF Regimen on the Expression of BCSG1 in Breast Cancer

Objective： To evaluate the efficacy of neoadjuvant chemotherapy and explore a sensitive and objective way in the evaluation of neoadjuvant chemotherapy, the pathological changes and BCSG1 expression were studied by pathological and immunohistochemical method in breast cancer patients with CAF neoadjuvant chemotherapy （Cyclophosphamide, Adriamycin and Fluorouracil, CAF） and those without at the same period. Methods： Specimens were obtained from 34 breast cancer patients receiving neoadjuvant CAF regimen chemotherapy （CAF group） and 110 breast cancer patients not receiving neoadjuvant chemotherapy （control group）. The BCSG1 expression was detected by SP immunohistochemistry. Correlation between BCSG1 expression and pathological response to CAF neoadjuvant chemotherapy was analyzed. Results： Overall response rate to neoadjuvant chemotherapy was 79.4%. The strong cytoplasm expression of BCSG1 was significantly lower in CAF group than in control group （29.4% vs. 64.5%, P〈0.01）. In CAF group, the positive cytoplasm expression in partial response （PR） （grade Ⅱ） cases was significantly lower than that in no response （NR） （grade Ⅲ） cases （P=0.002）. Conclusion： Neoadjuvant chemotherapy of CAF regimen could decrease the nuclear expression of BSCG1 in breast cancer.

Studies on mechanism of cis9,trans11-CLA and trans10,cis12-CLA inducing apoptosis of human breast cancer cell line MCF-7

Objective： The aim of the study was to explore the activities of cis9, trans11-CLA （C9, t11-CLA） and transl0, cis12-CLA （t10, c12-CLA） inhibiting tumor, and investigate their relationships with PPARy and apoptotic proteins, and mechanism of anti-cancer. Methods： The inhibitory rate, cell growth curve and apoptotic morphological observation of MCF-7 cells were obtained by MTT assay, trypan blue staining and Hoechst33342 fluorescence staining. The apoptotic rate and cell cycle were detected with flow cytometry. Transcriptional level of genes was detected with RT-PCR semi-quantitative method, and Western blot was performed to detect proteins levels. Results： The two CLA isomers could reduce cell proliferation （P 〈 0.05）, increase apoptotic rate （P 〈 0.05）, and increase obviously the transcriptional and protein levels of PPARy （P 〈 0.01）. The synchronism and correlation between the effects of CLA to PPARy and apoptotic proteins Bax, Bcl-2, Caspase 3 changes were found with the dose- and time-dependent manners. There was cooperative relation between the levels of PPARy and the rates of Bax/Bcl-2, Caspase 3 （small fragment） by experiments of PPARy inhibitor GW9662 and ligand Rosiglitazone. Conclusion： The apoptotic pathway of PPARy-Bcl-2-Caspase 3 signaling was found. The C9, t11-CLA and tl0, c12-CLA could inhibit MCF-7 cell proliferation and promote apoptosis via activating PPARy-Bcl-2-Caspase 3 pathway. CLA may be a kind of activator of PPARv.

Relationship between Full-Field Digital Mammographic Features and Clinicopathologic Characteristics in 176 Cases with Breast Cancer

OBJECTIVE Different mammographic features are probably predictive of different prognosis. However, ambiguity still exists in understanding the relationship between them. In resent years, digital mammography has been available for clinical use which has led to a revolution in the resolving of images and an increase in early-stage breast cancer detection. Based on the above knowledge, this study was performed to evaluate the relationship between full-field digital mammographic features and clinicopathologic characteristics in breast cancer. METHODS Digital mammograms of 176 patients with pathologically proven breast cancer were reviewed. Also, clinical and pathologic records （histological types and axillary lymph nodes status） were retrospectively examined. RESULTS Most of the patients with a solitary microcalcification were young women under the age of 50（84.4%）, but the majority of the patients with microcalcifications complicated by a mass were elderly women. Microcalcifications detected by mammography occurred frequently in ductal carcinoma in situ （28.1%） and in early invasive carcinoma （15.6%）. Breast cancers with expression of microcalcifications combined with a spiculate mass had a high metastatic rate of axillary lymph nodes （69.4%）. A high metastatic rate of axillary lymph nodes was also found in the patients with solitary worm-like microcalcifications （57.1%）, solitary spiculate mass （53.7%） and solitary non-worm-like microcalcifications （44.4%）. Simple worm-like microcalcifications accompanied with metastasis of 4 to 9 axillary lymph nodes occurred in 42.9% of the（6/14） cases. The patients with microcalcifications combined by a spiculate mass and with metastasis of 4 to 9 axillary lymph nodes accounted for 27.8% （10/36） of the cases, and those with metastases of 10 and over accounted for 16.7% （6/36）. CONCLUSION Solitary microcalcifications occur frequently in young women and are usually associated with early breast cancer. There is a close relationship between worm-like microcalcifications, a spiculate mass and positive metastases of axillary lymph nodes, which are an index of poor prognosis.

Papillary Adenocarcinoma of the Thyroid in Ectopic Intrathoracic Goiter:One Case Report

Case ReportA 58-year-old male patient presented to our department for surgical management of a right neck nodule and low fever over the past 2 weeks. Preoperative evaluation, which included chest CT scan, MRI and scintigraphy （^99mTc）, revealed round and clear boundary intrathoracic ectopic thyroid tissue at the right side of the anterior mediastinum and an enlarged lymph node in the right neck. The preoperative general image diagnosis concluded a malignant ectopic intrathoracic goiter （Figs. 1-3）. The lymph node biopsy confirmed a metastatic papillary adenocarcinoma of the thyroid.. The tumor was resected via a cervical collar incision （Fig.4）. Bilateral hemithyroidectomy and cervical lymph node dissection were also performed. We noticed that the intrathoracic thyroid was not connected to the cervical thyroid. Blood was supplied from the intrathoracic vessels, thereby establishing the diagnosis of an ectopic intrathoracic thyroid. Final pathologic diagnosis was a papillary adenocarcinoma of the thyroid in an ectopic intrathoarcic goiter with involved lymph node. The postoperative course was uneventful.

P─GLYCOPROTEIN EXPRESSION IN PRIMARY BREAST CANCER

In a retrospective study. liquid nitrogen preserved specimens from 50 women with primary breast cancer, who underwent surgery at the Beijing Institute for Cancer Research between June. 1986 and September, 1988, were investigated. All patients under this study were staged in TNM II or later, involved with axillary lymph node metastasis. and treated with systemic postoperative adjuvant chemotherapy. The median length of follow-up was 69 months. The expression of P-glycoprotein was investigated by means of immunohistochemistry, using a monoclonal antibody C219 specifically against P-glycoprotein and avidin -biotin peroxidase method. Posrtive staining for P-glycoprotein was found in 23(16%) of the 50 patients. The P-glycoprotein expression negative group fared better than the group that was P-glycoprotein posrtive in overall survival curves (p= 0. 0008 , by the generalized Wilcoxon test). The prognostic effect of P-glycoprotein expression remained statistically significant (p=0. 0007) after adjustment by multivariate analysis(Cox’s model)for other prognostic factors. It is demonstrated that P-glycoprotein expression is a significant and independent Predictor of postoperative survival in breast cancer patients. The results of the present study suggest that P-glycoprotein expression might also influence the bioiogical behavior of breast cancers.

Anti-breast cancer effects and mechanisms of Xihuang pill on human breast cancer cell lines

OBJECTIVE： To investigate the anti-breast cancer （BC） effects and mechanisms of action of Xihuang pill （XHP） by conducting in vitro experiments on hu- man BC cell lines. METHODS： Two human BC cell lines （MCF-7 and MDA- MB231） were cultured and treated with XHP. Cell viability was detected using the 3-（4, 5-Dimeth- ylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide （MTT） assay. Flow cytometry was used to measure the cell cycle and apoptosis. The cell cycle was ana- lyzed with propidium iodide staining. Apoptosis was evaluated using the Annexin V-fluorescein iso- thiocyanate/propidium iodide method. Western blotting was used to analyze the expression of es- trogen receptor （ER）-α and ER-13.RESULTS： XHP had growth-inhibitory effects on MCF-7 and MDA-MB231 cells with a half-maximal inhibitory concentration （IC50） of 10.14 mg/mL （MCF-7） and 8.98 mg/mL （MDA-MB231）. Apoptosis was induced to some extent. Certain changes in the ER were caused. Upregulation of ER-a protein was found in MCF-7 cells. ER-β expression in MDA-MB231 cells was increased. Cell-cycle arrest was not observed in the two BC cell lines. ER-β ex- pression in MCF-7 cells was unchanged. No ER-a ex- pression was shown in MDA-MB231 cells. CONCLUSION： These data suggest that XHP can af- fect cell viability and cause apoptosis, but that the cell cycle is not blocked. XHP has a certain impact on ER expression, but its mechanisms of action of anti-13C effects may not be due to regulation of ER expression.

Roles of miRNA and lncRNA in triple-negative breast cancer

Triple-negative breast cancer(TNBC)is currently the most malignant subtype of breast cancer without effective targeted therapies,which makes its pathogenesis an important target for research.A growing number of studies have shown that non-coding RNA(ncRNA),including microRNA(miRNA)and long non-coding RNA(lncRNA),plays a significant role in tumorigenesis.This review summarizes the roles of miRNA and lncRNA in the progression,diagnosis,and neoadjuvant chemotherapy of TNBC.Aberrantly expressed miRNA and lncRNA are listed according to their roles.Further,it describes the multiple mechanisms that lncRNA shows for regulating gene expression in the nucleus and cytoplasm,and more importantly,describes lnc RNA-regulated TNBC progression through complete combining with miRNA at the post-transcriptional level.Focusing on miRNA and lncRNA associated with TNBC can provide new insights for early diagnosis and treatment—they can be targeted in the future as a novel anticancer target of TNBC.