Doxorubicin resistance in breast cancer is mediated via the activation of FABP5/PPARγ and CaMKII signaling pathway.

Breast cancer is the most prevalent malignancy among women. Doxorubicin (Dox) resistance was one of the major obstacles to improving the clinical outcome of breast cancer patients. The purpose of this study was to investigate the relationship between the FABP signaling pathway and Dox resistance in breast cancer. The resistance property of MCF-7/ADR cells was evaluated employing CCK-8, Western blot (WB), and confocal microscopy techniques. The glycolipid metabolic properties of MCF-7 and MCF-7/ADR cells were identified using transmission electron microscopy, PAS, and Oil Red O staining. FABP5 and CaMKII expression levels were assessed through GEO and WB approaches. The intracellular calcium level was determined by flow cytometry. Clinical breast cancer patient's tumor tissues were evaluated by immunohistochemistry to determine FABP5 and p-CaMKII protein expression. In the presence or absence of FABP5 siRNA or the FABP5-specific inhibitor SBFI-26, Dox resistance was investigated utilizing CCK-8, WB, and colony formation methods, and intracellular calcium level was examined. The binding ability of Dox was explored by molecular docking analysis. The results indicated that the MCF-7/ADR cells we employed were Dox-resistant MCF-7 cells. FABP5 expression was considerably elevated in MCF-7/ADR cells compared to parent MCF-7 cells. FABP5 and p-CaMKII expression were increased in resistant patients than in sensitive individuals. Inhibition of the protein expression of FABP5 by siRNA or inhibitor increased Dox sensitivity in MCF-7/ADR cells and lowered intracellular calcium, PPARγ, and autophagy. Molecular docking results showed that FABP5 binds more powerfully to Dox than the known drug resistance-associated protein P-GP. In summary, the PPARγ and CaMKII axis mediated by FABP5 plays a crucial role in breast cancer chemoresistance. FABP5 is a potentially targetable protein and therapeutic biomarker for the treatment of Dox resistance in breast cancer.

Comprehensive analysis of the PTPN13 expression and its clinical implication in breast cancer.

Protein tyrosine phosphatases non-receptor 13 (PTPN13) could be a potential biomarker in breast cancer (BRCA), but its genetic variation and biological significance in BRCA remain undefined. Hereon, we comprehensively investigated the clinical implication of PTPN13 expression/gene mutation in BRCA. In our study, a total of 14 cases of triple-negative breast cancers (TNBC) treated with neoadjuvant therapy were enrolled, and post-operation TNBC tissues were collected for next-generation sequencing (NGS) analysis (422 genes including PTPN13). According to the disease-free survival (DFS) time, 14 TNBC patients were divided into Group A (long-DFS) and Group B (short-DFS). The NGS data displayed that the overall mutation rate of PTPN13 was 28.57% as the third highest mutated gene, and PTPN13 mutations appeared only in Group B with short-DFS. In addition, The Cancer Genome Atlas (TCGA) database demonstrated that PTPN13 was lower expressed in BRCA than in normal breast tissues. However, PTPN13 high expression was identified to be related to a favorable prognosis in BRCA using data from the Kaplan-Meier plotter. Moreover, Gene Set Enrichment Analysis (GSEA) revealed that PTPN13 is potentially involved in interferon signaling, JAK/STAT signaling, Wnt/ß-catenin signaling, PTEN pathway, and MAPK6/MAPK4 signaling in BRCA. This study provided evidence that PTPN13 might be a tumor suppressor gene and a potential molecular target for BRCA, and genetic mutation and/or low expression of PTPN13 predicted an unfavorable prognosis in BRCA. The anticancer effect and molecular mechanism of PTPN13 in BRCA may be associated with some tumor-related signaling pathways.

Clinicopathological characteristics and prognosis of breast cancer patients with lung cancer: A study based on 19,807 breast cancer patients.

This study was conducted to investigate the clinicopathological characteristics and prognosis of breast cancer and lung cancer (BC-LC) and provide a theoretical basis for the diagnosis and treatment of BC-LC in clinical work. A retrospective study was conducted on breast cancer (BC) patients in our center from September 2009 to November 2020. The patients were divided into the BC-LC group and the control group. The control group was matched with both, the age at diagnosis and the time of surgery (±1 year). The clinicopathological factors, overall survival (OS), and hazard ratios (HRs) were evaluated by SPSS. A total of 19,807 BC patients were identified, among whom 124 (0.6%) had lung cancer (LC). Larger BC tumor was the only independent risk factor (OR=2.454, p<0.001) for development of LC in BC patients. We found inferior survival in patients with synchronous versus metachronous BC-LC (p=0.008). We also identified combined with hypertension (HR=3.917, p=0.003) was an independent prognostic factor for inferior OS. Therefore, BC patients with larger tumors need close follow-up. Effective prevention and active treatment of hypertension can improve the OS of BC-LC patients.

Expression of BCRP/ABCG2 Protein in Invasive Breast Cancer and Response to Neoadjuvant Chemotherapy.

BACKGROUND: Breast cancer resistance protein (BCRP), or ATP-binding cassette subfamily G member 2 (ABCG2), is an ATP-binding cassette (ABC) transporter that mediates energy-dependent transport of substrate drugs out of the cell. Its overexpression may contribute to intrinsic drug resistance in vitro. However, the current literature has not yet clarified the clinical significance of BCRP/ABCG2 in invasive breast carcinoma. OBJECTIVES: The purpose of this study was to validate the expression of BCRP/ABCG2 in invasive breast carcinoma and its role in response to neoadjuvant chemotherapy. METHODS: In this study, a pretherapeutic core biopsy was performed in 222 patients. BCRP/ABCG2 expression in carcinoma tissue was measured by immunohistochemistry. BCRP/ABCG2 expression correlations with clinicopathological features, molecular subtypes, and therapy response after neoadjuvant chemotherapy were investigated. RESULTS: The results showed that BCRP/ABCG2 was expressed in different molecular subtypes. The proportions of patients with high BCRP/ABCG2 expression were similar in luminal A and luminal B tumors (luminal B, 80%; luminal A, 78%), compared with other molecular subtypes (triple-negative, 63%; HER2+, 58%, p = 0.05). BCRP/ABCG2 expression and the number of lymphatic metastases (p = 0.001) and tumor size (p = 0.011) demonstrated a statistically significant correlation. Low BCRP/ABCG2 expression was associated with an increased pathological complete response (pCR) rate of 38%, higher than the 19% in tumors with high BCRP/ABCG2 expression (p = 0.002). In multivariable analysis, BCRP/ABCG2 and hormone receptor (HR) expression were identified as independent risk factors of pCR (p = 0.003 and p = 0.013, respectively). CONCLUSIONS: BCRP/ABCG2 is highly expressed in HR-positive breast cancer. High BCRP/ABCG2 expression is associated with lymphatic metastasis, tumor size, and poor pCR. BCRP/ABCG2 may be a novel potential biomarker that can predict clinical progression and therapy response after neoadjuvant chemotherapy.

Machine learning-based random forest for predicting decreased quality of life in thyroid cancer patients after thyroidectomy.

OBJECTIVE: Decreased quality of life (QoL) in thyroid cancer patients after thyroidectomy is a common, but there is a lack of predictive methods for decreased QoL. This study aimed to construct a machine learning-based random forest for predicting decreased QoL in thyroid cancer patients 3 months after thyroidectomy. MATERIALS AND METHODS: Two hundred and eighty-six thyroid cancer patients after thyroidectomy were enrolled in this prospective cross-sectional study from November 2018 to June 2019, and were randomly assigned to training and validation cohorts at a ratio of 7:3. The European Organization for Research and Treatment of Cancer quality of life questionnaire version 3 (EORTC QLQ-C30) questionnaire was used to assess the QoL 3 months after thyroidectomy, and decreased QoL was defined as EORTC QLQ-C30 < 60 points. The random forest model was constructed for predicting decreased QoL in thyroid cancer patients after thyroidectomy. RESULTS: The mean QoL 3 months after thyroidectomy was 65.93 ± 9.00 with 21.33% (61/286) decreased QoL. The main manifestation is fatigue in symptom scales and social functioning dysfunction in functional scales. The top seven most important indices affecting QoL were clinical stage, marital status, histological type, age, nerve injury symptom, economic income and surgery type. For random forest prediction model, the areas under the curve in the training and validation courts were 0.834 and 0.897, respectively. CONCLUSION: The present study demonstrated that random forest model for predicting decreased QoL in thyroid cancer patients 3 months after thyroidectomy displayed relatively high accuracy. These findings should be applied clinically to optimise health care.

Prognostic Value of LRG1 in Breast Cancer: A Retrospective Study.

BACKGROUND: High expression of leucine-rich alpha-2-glycoprotein 1 (LRG1) is closely related to angiogenesis, which may play an important role in promoting invasion and metastasis. However, the current literature has yet to clarify the clinical significance of LRG1 in breast cancer. OBJECTIVES: The purpose of this work was to validate the correlation between LRG1 expression and prognosis in early breast cancer. METHODS: We utilized an LRG1 detection agent in 330 cases of early breast cancer. The correlation of LRG1 expression with clinicopathological features, patient recurrence, and survival was investigated. RESULTS: Compared with adjacent tissue samples, an elevated expression of LRG1 was observed in breast cancer samples. Moreover, LRG1 expression is associated with the number of lymphatic metastases and TNM pathological stage (p = 0.000, p = 0.000, respectively). For disease-free survival (DFS), the Kaplan-Meier curve indicated a poorer prognosis for the group with high LRG1 levels compared with the low LRG1 group (p = 0.000). A similar result was found for overall survival (OS; p = 0.000). The multivariate Cox regression indicated that LRG1 was still associated with DFS (HR 2.090, 95% CI 1.205-3.625, p = 0.009) and OS (HR 2.112, 95% CI 1.167-3.822, p = 0.013). The histological grade, TNM pathological stage, and molecular subtype were identified as independent risk factors affecting OS. CONCLUSION: In the malignant progression of breast cancer, high LRG1 levels are associated with lymphatic metastasis, histological grade, poor DFS, and poor OS. This study validates the use of LRG1 as a potential prognosis biomarker for early breast cancer.

Multi-center investigation of the clinical and pathological characteristics of inflammatory breast cancer based on Chinese Society of Breast Surgery (CSBrs-007).

BACKGROUND: Inflammatory breast cancer (IBC) is an aggressive type of cancer with poor prognosis and outcomes. This study aimed to investigate clinicopathological features, molecular characteristics, and treatments among Chinese patients diagnosed with IBC. METHODS: We collected data of 95 patients with IBC who were treated by members of the Chinese Society of Breast Surgery, from January 2017 to December 2018. The data, including demographic characteristics, pathological findings, surgical methods, systemic treatment plans, and follow-up, were obtained using a uniform electronic questionnaire. The clinicopathological features of different molecular types in patients without distant metastases were compared using the Kruskal-Wallis (H) test followed by post hoc analyses. RESULTS: Lymph node metastasis was noted in 75.8% of all patients, while distant metastasis was noted in 21.4%. Pathological findings indicated invasive ductal and lobular carcinomas in 86.8% and 5.3% of cases, respectively. Hormone receptor-positive (HR+)/human epidermal growth factor receptor 2-negative (HER2-) (41.5%) and HR-/HER2+ (20.1%) were the most common biologic subtypes, followed by HR+/HER2+ (19.1%) and HR-/HER2- (19.1%). Stage III IBC was treated via pre-operative neoadjuvant chemotherapy in 87.7% of the cases, predominantly using anthracycline and taxanes. A total of 91.9% of patients underwent surgical treatment. Among them, 77.0% of the patients underwent modified radical mastectomy, 8.1% of whom also underwent immediate breast reconstruction. The Kruskal-Wallis test revealed that the efficacy of chemotherapy significantly differed among those with HR+/HER2- and HR-/HER2- tumors (adjusted P = 0.008), and Ki-67 expression significantly differed in HR-/HER2+ and HR+/HER2+ molecular subtypes (adjusted P = 0.008). CONCLUSION: Our study provides novel insight into clinicopathological characteristics and treatment status among patients with IBC in China, and might provide a direction and basis for further studies. TRIAL REGISTRATION: chictr.org.cn, No. ChiCTR1900027179; http://www.chictr.org.cn/showprojen.aspx?proj=45030.

p-Hydroxylcinnamaldehyde from cochinchinamomordica seed reverses resistance to TRAIL in human oesophageal squamous cell carcinoma via the activation of the p38 mitogen-activated protein kinase signalling pathway.

BACKGROUND: Our previous studies have showed that p-Hydroxylcinnamaldehyde (CMSP) could induce the differentiation of ESCC cells via the cAMP-RhoA-MAPK signalling pathway, which suggests a new potential strategy for ESCC treatment. Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potent inducer of apoptosis in several tumour cells by binding to the death receptors DR4 and DR5. However, TRAIL has little effect on oesophageal squamous cell carcinoma (ESCC) cells due to the loss of the receptors. The present study determined the effect of CMSP, the firstly found chemical constituent of Cochinchinamomordica seed (CMS), on TRAIL-induced apoptosis and its mechanism in ESCC cells. METHODS: MTS assays were performed to examine the CMSP- and TRAIL-mediated inhibition of ESCC cell growth. Flow cytometry and Hoechst 33258 staining assays were used to detect apoptosis in ESCC cells treated with CMSP combined with TRAIL. Western blotting was used to determine the effect of CMSP on the expression of p38, p-p38, DR4, DR5, Bid and caspase-3/8 in ESCC cells treated with CMSP combined with TRAIL. Additionally, immunodeficient Balb-c/null mouse model was used to determine the chemotherapeutic efficacy of CMSP and TRAIL against ESCC tumour xenograft growth in vivo. RESULTS: We found that the combination of CMSP and TRAIL had a greater inhibitory effect on ESCC cell viability in vitro than CMSP or TRAIL alone. CMSP enhanced the TRAIL-induced apoptosis in ESCC cells by upregulating the expression of DR4 and DR5 via the p38 MAPK signalling pathway. Furthermore, the increased expression of DR4 and DR5 upon TRAIL-induced apoptosis in ESCC cells was mediated at least in part by subsequent caspase-3 and caspase-8 activation. Moreover, the in vivo model showed that tumour growth was significantly slower in CMSP and TRAIL combination-treated mice than in mice treated with CMSP or TRAIL alone. CONCLUSION: Taken together, our findings indicate that CMSP as an extract from TCM, might be as a potential sensitizer of TRAIL and thus provide a novel strategy for the clinical treatment of ESCC.

Association of p53 expression with poor prognosis in patients with triple-negative breast invasive ductal carcinoma.

TP53 gene is mutated in approximately 80% of triple-negative breast cancer (TNBC). However, the prognostic significance of immunohistochemical (IHC)-detected p53 protein expression remains controversial in TNBC. In this study, we retrospectively analyzed the association between IHC-detected p53 expression and the prognosis in a cohort of 278 patients with stage I-III triple-negative breast invasive ductal carcinoma (IDC), who received surgery at the department of breast surgery in the Fourth Hospital of Hebei Medical University from 2010-01 to 2012-12. We found a positive expression ratio of IHC-detected p53 in triple-negative breast IDC of 58.6% (163/278). Furthermore, levels of expression were significantly associated with vessel tumor emboli and higher histologic grade (P = .038, P = .043, respectively), with the highest expression level observed in G3 breast cancer (64.7%). Additionally, Kaplan-Meier analysis showed that p53 expression indicated worse overall survival (OS) in the whole cohort (79.6% vs 89.6%, Log-rank test P = .025) as well as in stratified prognostic stage II patients (90.8% vs 100%, Log-rank test P = .027). The mortality risk of p53 expression patients was 2.22 times higher than that of p53 negative patients (HR: 2.222; 95%CI: 1.147-4.308). In addition, p53 expression was also associated with poor disease-free survival (DFS) (76.7% vs 86.8%, P = .020). Cox proportional hazard ratio model showed p53 expression was an independent risk factor for OS (P = .018) and DFS (P = .018) after controlling for tumor size, lymph node status, and vessel tumor emboli. Altogether, our data showed that IHC-detected p53 expression is a promising prognostic candidate for poor survival in triple-negative breast IDC patients. However, more studies are needed to determine if p53 may be applied to clinical practice as a biomarker and/or novel therapeutic target for TNBC.

[Biological characteristics of CD55(hig); side population in breast cancer cell line MCF-7].

AIM: To determine whether the CD55(hig); expression (CD55(hig);) cells in side population (SP) of the cell line MCF-7 possess characteristics of cancer stem cells. METHODS: Breast cancer cell line MCF-7 was cultured and the nucleic acid dye Hoechst33342 and Verapami were added. Flow cytometry (FCM) was employed to isolate the cells of SP and mian population (MP), the cells were then labeled with CD55 mAb; mean values of fluorescence intensity of CD55 in SP and MP cells were measured. CD55 mAb was used to mark the unsorted MCF-7 cells, the proportion of CD55(hig); cells was determined, and then CD55(hig); cells were sorted and collected, to observe biological characteristics, such as cell morphology, adherence rate, colony formation and cell cycle distribution as well as nude mice implantation. RESULTS: Mean value of fluorescence intensity of CD55 in SP cells was 100.85±4.57, and mean value of fluorescence intensity of CD55 in MP cells was 50.51±4.75; the proportion of CD55(hig); cell in MCF-7 cells was 2.12%; adherent rate of CD55(hig); cells in 24 h was lower than that in CD55(low); cells and 24 h after inoculation adherent rates of both cells had no significant difference (P>0.05); CD55(hig); cells were mostly spherical and kept in a suspended state at 12 hours after inoculation and culture; CD55(hig); cells had the ability of colony formation, the clone-forming rate of CD55(hig); cells was (20.04±1.07)% when the cells were cultured for one week, it was lower than the rate of (27.14±1.07)% in CD55(low); cells (P<0.01); the ratio of G0/G1 resting cells in CD55(hig); cells was (85.4±3.37)% which was higher than that in CD55(low); cells (58.6±2.55)% and in MCF-7 cells (70.73±4.21)%, which had a statistically significant difference (P<0.05). All nude mice implanted with CD55(hig); cells developed the tumor, and the pathological examination of the transplanted tumor had the properties of malignant cells. CONCLUSION: A few CD55(hig); cells were found in breast cancer MCF-7 cells, and most of CD55(hig); cells were quiescent, non-adherent as well as being in a suspended state and being hereditary after cloning, so CD55(hig); cells had biological characteristics of cancer stem cells.

[Induction of specific CTL by DC from axillary lymph node of patients with breast cancer].

AIM: To investigate the capability of the dendritic cells (DC) from axillary draining lymph node of patients with breast cancer to induce the antigen specific CTL. METHODS: The mononuclear cells were isolated from axillary draining lymph node, and the adherent cells were cultured with rhGM-CSF and rhIL-4 to induce DC. The non-adherent cells were cultured with IL-2 to induce tumor draining lymph node cells(TDLNC). DCs stimulated by the autoallergic breast cancer freeze-thawing antigen were co-cultured with the obtained TDLNC to induce tumor antigen specific CTL. RESULTS: The positive percentage of CD1a, CD83, CD86 on DC from freshly isolated mononuclear cells in axillary draining lymph node was 11.0+/-2.3, 26.6+/-5.2 and 33.0+/-6.1, respectively. After co-cultured with rhGM-CSF, rhIL-4, breast cancer freeze-thawing antigen and TNF-alpha, the percentage of CD1a, CD83, CD86 was increased to 50.2+/-5.7, 60.5+/-16.5 and 56.2+/-16.4, respectively, (P<0.01). The percentage of CD8(+) T cells in TDLNC increased from 32.8+/-3.2 to 62.5+/-2.5 after co-cultured with DC-Ag (P<0.01). CONCLUSION: Typical DCs can be induced from the mononuclear cells in axillary draining lymph node after stimulated with cytokine (rhGM-CSF, rhIL-4 and TNF-alpha), which possess strong antigen presentation function and can stimulate TDLNC to proliferate and differentiate into antigen specific CTL.