Evaluación de la expresión de PD-L1 e infiltración linfocitaria intratumoral en carcinoma de mama invasivo triple negativo / Evaluation of PD-L1 expression and intratumoral lym phocytic infiltration in triple-negative invasive breast car cinoma

Resumen Introducción : El endofenotipo de cáncer de mama triple negativo (TNBC) es uno de los menos frecuentes y sin diana terapéutica, por tanto, se plantea estudiar la correlación del punto de control inmunológico PD-L1 con el establecimiento de microambiente tumoral evaluado por la infiltración linfocitaria intratumoral estromal (TILs) y su importancia en la práctica clínica. Métodos : Se realizó un estudio retrospectivo de casos y controles, con 31 casos de carcinoma infiltrante de la mama triple negativo y 57 controles no pareados de endofenotipo Luminal A, Luminal B y HER-2 atendidos en un año. Se evaluaron las variables: tipo y grado his tológico, expresión PD-L1 con el clon 22C3, TILs, invasión linfovascular, tamaño tumoral, compromiso de ganglios linfáticos y metástasis. El análisis estadístico se ejecutó con la prueba de chi cuadrado y prueba de coeficiente de correlación de Spearman. Resultados : Se encontró una correlación negativa estadísticamente significativa entre TILs y PD-L1 (rho - 0.106, p 0.025), indicando que a mayor expresión de PD-L1, es menor la infiltración linfocitaria intratumo ral. En los grupos de TILs B (10-40% TILs) y C (40-90% TILs) donde se presenta marcado infiltrado inflamatorio intratumoral se evidenció mayor número de pacientes negativos para PD-L1 (CPS <10) con 16 y 10 casos res pectivamente. Para los casos TNBC se logró identificar un coeficiente de asociación negativa (rho -0.378) y con significancia estadística (p 0.01). Discusión : Se estableció la asociación de TNBC, TILs y expresión de PDL1, lo cual es importante para la instau ración de terapias diana y el desarrollo de la medicina de precisión.

Abstract Introduction : Triple negative breast cancer endophe notype (TNBC) is one of the least frequent and without therapeutic target; therefore we propose to study the correlation of PD-L1 immune checkpoint with the es tablishment of tumor microenvironment assessed by intratumoral stromal lymphocyte infiltration (TILS) and its importance in clinical practice. Methods : A retrospective case-control study was performed, with 31 cases of triple-negative infiltrat ing breast carcinoma and 57 unmatched controls of Luminal A, Luminal B and HER-2 endophenotype seen in one year. The following variables were evaluated: histologic type and grade, PD-L1 expression with clone 22C3, TILS, lymphovascular invasion, tumor size, lymph node involvement and metastasis. Statistical analysis was performed with the chi-square test and Spearman correlation coefficient test. Results : a statistically significant negative correlation was found between TILS and PD-L1 (rho - 0.106, p 0.025), indicating that the higher the expression of PD-L1, the lower the intratumoral lymphocytic infiltration. In the TILS B (10-40% TILS) and C (40-90% TILS) groups where there was a marked intratumoral inflammatory infiltrate, a greater number of patients were negative for PD-L1 (CPS <10) with 16 and 10 cases, respectively. For TNBC cases a negative association coefficient was identified (rho -0.378) with statistical significance (p 0.01). Discussion : The association between TNBC, TILS and PDL1 expression was established, which is important for the establishment of target therapies and the develop ment of precision medicine.

Exploring mechanism of Huayu Pills combined with immunocheckpoint inhibitors in treating triple-negative breast cancer based on network pharmacology / 国际中医中药杂志

Objective:To explore the potential mechanism of Huayu Pills in the treatment of TNBC (triple-negative breast cancer) using network pharmacology.Methods:TCMSP (Traditional Chinese Medicine Systems Pharmacology), PubChem, STITCH and SwissTargetPrediction databases were used to analyze the potential targets of Huayu Pills. TNBC disease targets were screened based on the TCGA (The Cancer Genome Atlas) database. The drug-disease mapping target was constructed by PPI network, key target screening and module analysis, and the DAVID database was used for GO function annotation and KEGG signal pathway enrichment analysis. SD rats were orally administered 3.94 g/kg of Huayu Pills decoction for 4 days to prepare medicated serum. HUVEC cells were randomly divided into a control group and an experimental group using a random number table method. The experimental group received intervention with Huayu Pills containing serum for 24 hours, inoculated in matrigel's solidified 48 well plate and HUVEC angiogenesis was observed 3 hours later.Results:130 possible targets of Huayu Pills in the treatment of TNBC were obtained. VEGFA is the core target. The cascade of angiogenesis, hypoxia, and blood coagulation may be the main functions and key signals of Huayu Pills in the treatment of TNBC. In vitro studies have shown that serum containing Huayu Pills can promote the normalization of tumor blood vessels in HUVEC cells.Conclusion:Huayu Pills may promote tumor vascular normalization (angiogenesis, hypoxia, coagulation cascade reactions) through VEGF targets, and can assist immune checkpoint inhibitors in the treatment of TNBC.

A molecular approach to triple-negative breast cancer: targeting the Notch signaling pathway

ABSTRACT Introduction Triple-negative breast cancer is an aggressive subtype of breast cancer characterized by the absence of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 expression. This phenotype renders triple-negative breast cancer cells refractory to conventional therapies, resulting in poor clinical outcomes and an urgent need for novel therapeutic approaches. Recent studies have implicated dysregulation of the Notch receptor signaling pathway in the development and progression of triple-negative breast cancer. Objective This study aimed to conduct a comprehensive literature review to identify potential therapeutic targets of the Notch pathway. Our analysis focused on the upstream and downstream components of this pathway to identify potential therapeutic targets. Results Modulating the Notch signaling pathway may represent a promising therapeutic strategy to treat triple-negative breast cancer. Several potential therapeutic targets within this pathway are in the early stages of development, including upstream (such as Notch ligands) and downstream (including specific molecules involved in triple-negative breast cancer growth). These targets represent potential avenues for therapeutic intervention in triple-negative breast cancer. Comments Additional research specifically addressing issues related to toxicity and improving drug delivery methods is critical for the successful translation of these potential therapeutic targets into effective treatments for patients with triple-negative breast cancer.

Real-world data on adjuvant capecitabine after standard neoadjuvant chemotherapy for triple negative breast cancer

Abstract Objective Neoadjuvant chemotherapy (NACT) has become the standard of care for patients with triple-negative breast cancer (TNBC) with tumors > 1 cm or positive axillary nodes. Pathologic complete response (pCR) has been used as an endpoint to select patients for treatment scaling. This study aimed to examine the benefit of adding adjuvant capecitabine for TNBC patients who did not achieve pCR after standard NACT in a real-world scenario. Methods This retrospective cohort study included all patients with TNBC who underwent NACT between 2010 and 2020. Clinicopathological data were obtained from the patient records. Univariate and multivariate analyses were conducted at the 5 years follow-up period. Results We included 153 patients, more than half of whom had stage III (58.2%) and high-grade tumors (60.8%). The overall pCR rate was 34.6%, and 41% of the patients with residual disease received adjuvant capecitabine. Disease-specific survival (DSS) among the patients who achieved pCR was significantly higher (p<0.0001). Residual disease after NACT was associated with detrimental effects on DSS. In this cohort, we did not observe any survival benefit of adding adjuvant capecitabine for patients with TNBC subjected to NACT who did not achieve pCR (p=0.52). Conclusion Our study failed to demonstrate a survival benefit of extended capecitabine therapy in patients with TNBC with residual disease after NACT. More studies are warranted to better understand the indication of systemic treatment escalation in this scenario.

Carcinoma de mama triple negativo. Estudio anatomo-estadístico / Triple- negative breast cancer. An anatomical and statistical study

Introducción: El carcinoma de mama triple negativo se asocia a un comportamiento biológico más agresivo y de desfavorable pronóstico. Objetivo: Determinar la incidencia del subtipo molecular triple negativo en carcinomas mamarios y su relación con otras variables clínico-patológicas de valor pronóstico. Método: Se realizó un estudio descriptivo y retrospectivo, en el Hospital Universitario Docente «Celestino Hernández Robau» de Villa Clara, en el período comprendido de enero de 2017 a junio de 2019, con el fin de determinar la incidencia de los tumores triples negativos y su relación con las variables edad, talla tumoral, tipo y grado histológicos e índice de proliferación. Resultados: Se determinó la incidencia del subtipo molecular triple negativo en carcinomas mamarios y su relación con las formas histológicas moderada y poco diferenciadas. Conclusiones: El subtipo molecular triple negativo en carcinomas mamarios está asociado con frecuencia a: la edad posmenopáusica, el tipo histológico ductal, el grado histológico alto, altos índices de Ki-67 y talla tumoral mayor de 2 cm.

Introduction: triple-negative breast cancer has a more aggressive biological behaviour and is associated with an unfavourable prognosis. Objective: to determine the incidence of triple- negative breast cancer molecular subtypes and its relationship with other clinical and pathological variables of prognostic value. Methods: a descriptive and retrospective study was carried out at "Celestino Hernández Robau" University Teaching Hospital from Villa Clara between January 2017 and June 2019 in order to determine the incidence of triple- negative tumors and its relationship with the variables: age, tumor size, histological type and grade as well as proliferative index. Results: the incidence of triple- negative breast cancer molecular subtype and its relationship with moderate and poorly differentiated histological forms were determined. Conclusions: triple- negative breast cancer molecular subtype is frequently associated with postmenopausal age, ductal histological type, high histological grade, high Ki-67 indices and tumor size greater than 2 cm.

Research progress of p53 protein in the development mechanism of triple negative breast cancer / 中国医师杂志

Tumor suppressor gene p53 plays an important role in regulating cell cycle, controlling apoptosis and repairing damaged DNA. Mutation of this gene is closely related to the occurrence, development and drug resistance of various tumors. The mutant p53 protein is closely related to the growth and metastasis of triple negative breast cancer (TNBC) with higher malignancy and higher risk of metastasis. This paper expounds the mechanism of p53 protein participating in the occurrence, development and metastasis of TNBC, introduces the effect of interfering with mouse dual-microbody gene 2 (MDM2), activated T cell nuclear factor 1 (NFAT1) and other proteins on p53, as well as small molecular targeted drugs closely related to p53 protein, and provides a new direction and theoretical basis for targeted treatment of TNBC.

Effect of transcription factor NFIB on cell proliferation and invasion in breast cancer / 中国医师杂志

Objective:To investigate the effect of transcription factor nuclear factor IB (NFIB) on cell proliferation and invasion in breast cancer.Methods:The lentivirus pLKO.1-shNFIB plasmid was constructed, packaged and infected with human estrogen receptor positive (ER + ) breast cancer cell line MCF-7 and triple-negative breast cancer (TNBC) cell line MDA-MB-231, respectively, NFIB was stably knocked down and verified by Western blot method; Cell count test (CCK-8) and clone formation test were used to investigate the effect of knockdown NFIB on the growth and proliferation of breast cancer cells; The transwell experiment and Western blot method were performed to detect the expression of epithelial mesenchymal transition protein markers. The effect of knockdown NFIB on the invasive ability of triple-negative breast cancer cells was explored; Kaplan-Meier survival was used to analyze web data (http: //kmplot.com/analysis/) to explore the effect of NFIB on the prognosis of ER + breast cancer and triple-negative breast cancer patients. Results:In MCF-7 and MDA-MB-231 breast cancer cells, knocking down NFIB inhibited cell growth and proliferation; In triple-negative breast cancer MDA-MB-231 cells, knocking down NFIB promoted the expression of interstitial marker fibronectin and promoted cell invasion; The lower the expression of NFIB, the worse the prognosis of triple negative breast cancer patients, while the expression of NFIB had no effect on the prognosis of ER + breast cancer patients. Conclusions:Knocking down NFIB inhibits the proliferation of MCF-7 cells, and the expression level of NFIB is not related to the prognosis of ER + breast cancer patients; Knocking down NFIB inhibits the proliferation of MDA-MB-231 cells but promotes their invasion; The low expression of NFIB is associated with the poor prognosis of triple-negative breast cancer patients.

Effect of fritinib on angiogenesis, tumor growth and IRE1-ASK1-JNK pathway in triple negative breast cancer / 国际肿瘤学杂志

Objective:To investigate the effects of fritinib on angiogenesis, tumor growth and inositol requiring enzyme 1 (IRE1) -apoptosis signal regulating kinase 1 (ASK1) -c-Jun N-terminal kinase (JNK) pathway in triple negative breast cancer.Methods:Triple negative breast cancer cells MDA-MB-231 were taken and divided into normal saline (NS) group, low-dose fritinib (LD) group, medium-dose fritinib (MD) group and high-dose fritinib (HD) group. NS group was added with 100 μmol/L normal saline. LD group, MD group and HD group were added with 25, 50 and 100 μmol/L fritinib, respectively. Cell proliferation was detected by MTT assay, cell apoptosis was detected by flow cytometry, the ability of cells to form mimicry vessels was observed by three-dimensional cell culture, and the related indexes of angiogenesis and IRE1-ASK1-JNK pathway were detected by Western blotting. Twenty triple-negative breast cancer rat models were divided into control group and experimental group by random number table method, with 10 rats in each group. The control group was given normal saline gavage and the experimental group was given 100 μmol/L fritinib gavage. The tumor growth of rats in the two groups was observed and recorded.Results:The 48 h cell proliferation rates of NS group, LD group, MD group and HD group were (85.44±5.58) %, (73.24±4.95) %, (61.53±4.07) % and (50.23±2.97) %, respectively ( F=4.01, P=0.002). Compared with the NS group, the cell proliferation rate in LD, MD and HD groups was significantly decreased in a dose-dependent manner (all P<0.05). The apoptosis rates of NS group, LD group, MD group and HD group were (3.41±0.39) %, (18.75±1.94) %, (24.97±2.58) % and (38.62±3.27) %, respectively ( F=18.99, P<0.001). Compared with the NS group, the apoptosis rate of LD, MD and HD groups was significantly increased in a dose-dependent manner (all P<0.05). The number of mimicry vessels in NS group, LD group, MD group and HD group was 19.58±2.11, 15.67±2.02, 11.57±1.73 and 5.20±1.23, respectively ( F=3.28, P=0.008). Compared with the NS group, the number of mimicry vessels in LD group, MD group and HD group was significantly reduced. The results were dose-dependent (all P<0.05). vascular endothelial growth factor (VEGF) protein expression in NS group, LD group, MD group and HD group was 2.36±0.21, 1.79±0.17, 1.48±0.14 and 0.94±0.10, respectively ( F=5.17, P<0.001). The expression of IRE1 protein was 1.18±0.12, 1.67±0.18, 2.03±0.24 and 2.39±0.28, respectively ( F=5.55, P<0.001). The expression of ASK1 protein was 1.09±0.11, 1.46±0.13, 1.81±0.18, 2.33±0.21 ( F=5.32, P<0.001), respectively. JNK protein expression was 1.01±0.09, 1.48±0.14, 1.86±0.21 and 2.28±0.24, respectively ( F=6.92, P<0.001). Compared with the NS group, VEGF protein expression in LD, MD and HD groups was significantly decreased, and the expressions of IRE1, ASK1 and JNK were significantly increased in a dose-dependent manner (all P<0.05). Compared with the control group, the tumor weight and volume of the experimental group were significantly decreased [ (0.55±0.10) g vs. (1.37±0.15) g, t=14.38, P<0.001; (77.39±3.21) mm 3vs. (118.26±5.34) mm 3, t=20.74, P<0.001], tumor inhibition rate was significantly increased [ (71.23±3.85) % vs. (32.56±3.08) %, t=24.80, P<0.001]. Conclusion:Fritinib has an inhibitory effect on the activity of triple negative breast cancer cells, which can significantly reduce their angiogenesis and inhibit tumor growth. Moreover, it is related to the activation of IRE1-ASK1-JNK pathway.

Progress of immunotherapy-based strategy in triple-negative breast cancer / 国际肿瘤学杂志

Triple-negative breast cancer (TNBC) represents a distinct subtype of breast cancer, characterized by unique clinical traits including early lung metastasis, elevated recurrence rates, and diminished survival prospects. Owing to the lack of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 expression, concrete therapeutic targets remain elusive, thereby confining available clinical treatment methods. In the context of advanced TNBC, chemotherapy remains the predominant therapeutic approach. In recent years, with the in-depth study of tumor microenvironment, new immunotherapy targets have been discovered one after another. Thus, immunotherapy-based combined therapy strategies have brought new hope in patients with advanced TNBC.

Remodeling of tumor stroma combined with photothermal therapy in the treatment of triple-negative breast cancer / 中华肿瘤杂志

Objective: Polyethylene glycol-modified gold nanostar particles (GNS-PEG) were constructed to investigate whether the degradation of extracellular matrix in triple-negative breast cancer could improve the tumor delivery of GNS-PEG and enhance the efficacy of photothermal therapy. Methods: GNS-PEG were constructed and characterized for physicochemical properties as well as photothermal properties. At the cellular level, the cytotoxicity of halofuginone (HF) and the effect of photothermal therapy were detected. Mouse model of triple negative breast cancer was established by subcutaneous inoculation of 4T1 cells in BALB/c nude mice. Five injections of HF were given via tail vein (HF group), and tumor sections were stained with Masson stain and immunohistochemical staining for transforming growth factor β1 (TGFβ1), α-smooth muscle actin (α-SMA) and CD31 to observe the effect of tumor stromal degradation. Five injections of HF via tail vein followed by GNS-PEG (HF+ GNS-PEG group) were applied to determine the content of gold in tumor tissues by inductively coupled plasma mass spectrometry. The tumor sites of the mice in the GNS-PEG and HF+ GNS-PEG groups were irradiated with NIR laser and the temperature changes were recorded with an IR camera. The tumour growth and weight changes of mice in each group were observed. Ki-67 immunohistochemical staining, TdT-mediated dUTP nick-end labeling and HE staining were performed on tumor tissue sections from each group to observe tumor proliferation, apoptosis and necrosis. HE staining was performed on heart, liver, spleen, lung and kidney tissues from each group to observe the morphological changes of cells. Results: GNS-PEG nanoparticles showed a multi-branched structure with a particle size of 73.5±1.4 nm. The absorption peak of GNS was 810 nm, which is in the near infrared region. The photothermal conversion rate of GNS-PEG was up to 79.3%, and the photothermal effect could be controlled by the laser energy. HF has a concentration-dependent cytotoxicity, with a cell survival rate being as low as (22.8±2.6)% at HF concentration of up to 1 000 nmol/L. The photothermal effect of GNS-PEG was significant in killing tumor cells, with a cell survival rate of (32.7±5.2)% at the concentration of 25 pmol/L. The collagen area fraction, TGFβ1 integrated optical density and α-SMA integrated optical density in the tumor tissues of mice in the HF group were (2.1±0.2)%, 3.1±0.4 and 5.2±1.9, respectively, which were lower than those of the control group (all P<0.01), and the vessel diameter was 8.6±2.9 μm, which was higher than that of the control group (P<0.05). In the HF+ GNS-PEG group, the concentration of gold in tissues was 52.4 μg/g, higher than that in the GNS-PEG group (15.9 μg/g, P<0.05). After laser irradiation, the temperature of the tumor site in the HF+ GNS-PEG group was significantly higher than that in the GNS-PEG group. At the 4th minute, the temperatures of the tumor site in the GNS-PEG and HF+ GNS-PEG groups were 51.5 ℃ and 57.7 ℃ respectively; the tumor volume in the HF+ GNS-PEG group was effectively suppressed. The body weights of the mice in each group did not change significantly during the monitoring period. No significant abnormalities were observed in the main organs of the mice in the GNS-PEG group, but some hepatocytes in the HF and HF+ GNS-PEG groups showed edema and degeneration. Conclusion: The remodeling of extracellular matrix in triple-negative breast cancer could significantly improve the intratumoral delivery of GNS-PEG and thus achieve better photothermal therapy effect.