Nischarin inhibits the epithelial-mesenchymal transition process and angiogenesis in breast cancer cells by inactivating FAK/ERK signaling pathway via EGF like repeats and discoidin domains 3.

BACKGROUND: Our previous study has demonstrated that Nischarin (NISCH) exerts its antitumor effects in breast cancer (BC) by suppressing cell migration and invasion. This study aims to explore the underlying mechanism through which NISCH functions in BC. METHODS AND RESULTS: The relevance between EGF Like Repeats and Discoidin Domains 3 (EDIL3) mRNA expression and the overall survival of tumor patients was depicted by the Kaplan-Meier curve. The findings revealed that overexpressed NISCH attenuated cell motility and colony-forming capacities of Hs578T cells, yet silenced NISCH in MDA-MB-231 cells led to contrasting results. Western blot (WB) analysis indicated that overexpression of NISCH significantly down-regulated the Vimentin and Slug expression, and inactivated the FAK/ERK signaling pathway. RNA sequencing (RNA-seq) was performed in NISCH-overexpressed Hs578T cells and the control cells to analyze differentially expressed genes (DeGs), and the results showed a significant down-regulation of EDIL3 mRNA level upon overexpression of NISCH. Subsequent functional analyses demonstrated that overexpression of EDIL3 attenuated the inhibitory effect of NISCH on cell migration, invasion, colony formation, and tube formation. CONCLUSION: In summary, our finding preliminarily revealed that NISCH inhibits the epithelial-mesenchymal transition (EMT) process and angiogenesis in BC cells by down-regulating EDIL3 to inactivate the FAK/ERK signaling pathway, thereby suppressing the progression of BC. Our results hold promise for contributing to the deep understanding of BC pathogenesis and identifying new therapeutic strategies for clinical application.

Disparities in quality of life among patients with breast cancer based on surgical methods: a cross-sectional prospective study.

To determine the impact of breast conservation on quality of life and identify treatment-related and other demographic factors associated with post-breast cancer treatment quality of life. A prospective study was conducted on 392 women who underwent breast cancer surgery at Hangzhou Cancer Hospital from January 1, 2013, to December 31, 2022. Operable breast cancer patients who had completed all treatments except endocrine therapy were included. Patients with tumor recurrence/metastasis, bilateral or male breast cancer, and other primary malignancies were excluded. After enrollment, patients were asked to complete the BREAST-Q scale, and their pathological and medical records were reviewed. Analysis of variance was used to compare the quality of life scores among the groups. Univariate and multivariate linear regression analyses were performed to identify independent factors associated with quality of life scores in different domains. Participants completed the BREAST-Q scale at a median of 4.6 years after surgery. Quality of life scores varied based on the therapeutic strategy. Breast conservation has significant advantages over mastectomy in terms of breast satisfaction, psychosocial, and sexual well-being. Compared to oncoplastic breast-conserving surgery, mastectomy was independently associated with decreased breast satisfaction, psychosocial, and sexual well-being, while conventional breast-conserving surgery showed comparable outcomes to oncoplastic breast-conserving surgery in terms of these factors. Breast conservation leads to an improvement in quality of life compared to mastectomy. Oncoplastic breast-conserving surgery does not lead to a decrease in quality of life compared to conventional breast-conserving surgery and offers better outcomes compared to mastectomy.

Lateral thoracic adipofascial flaps in the reconstruction of defects after breast-conserving surgery in breasts with ptosis mild ptosis or without ptosis.

OBJECTIVE: To study the feasibility, safety, and effectiveness of lateral thoracic adipofascial flaps in reconstructing the defects following breast-conserving surgery (BCS) in breasts with either no ptosis or mild ptosis. METHODS: 37 female patients who underwent BCS and lateral thoracic adipofascial flap breast reconstruction between June 2020 and July 2022 were analysed. Surgery-related complications, intraoperative positive margin, local recurrence, and cosmetic outcome were assessed. RESULTS: Three local complications occurred in patients, all of which were cured by conservative treatment. Additionally, four patients had intraoperative positive margins. After a median follow-up period of 17.5 months, none of the patients showed local recurrence. All patients achieved a satisfactory breast shape. Further, patients without ptosis achieved good volume and symmetry. However, the breast symmetry was not satisfactory for patients with ptosis. CONCLUSION: It is reliable and effective to use the lateral thoracic adipofascial flaps to reconstruct the defects after BCS when the breast is not ptotic and the lesions are located in the lateral and central quadrants.

KMT2D Regulates the NCOA6/THRB Signal Axis through Epigenetic Modification to Promote the Migration and Invasion of Papillary Thyroid Cancer.

BACKGROUND: The histone lysine methyltransferase Histone-lysine N-methytransferase 2D (KMT2D) is a common mutated gene in a variety of cancers, including papillary thyroid cancer (PTC). However, the mechanism of KMT2D on the progression of PTC remains unclear. METHODS: In this study, quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting were used to evaluate KMT2D expression between human normal cell (Nthy-ori 3-1) and PTC cells (TPC1, IHH-4 and BCPAP). Proliferation, migration and invasion of TPC1, IHH-4 and BCPAP were assessed by Cell Counting Kit-8 (CCK-8), Wound-healing assay and Transwell assay. The mechanism of KMT2D on thyroid papillary cancer was explored with Chromatin immunoprecipitation assay (ChIP), qRT-PCR and Western blotting. RESULTS: The expression of KMT2D in PTC cells was significantly increased. Downregulation of KMT2D significantly decreased the proliferation, migration and invasion of PTC cells, which was correlated with decreased expression levels of H3K4me2, H3K9me2, NCOA6 and THRB. Meanwhile, ChIP assay demonstrated that KMT2D was associated with NCOA6. CONCLUSIONS: Study have shown that the downregulation of KMT2D reduces proliferation, migration and invasion of thyroid papillary carcinoma cells through epigenetic modification of NCOA6/THRB signal axis. These results provide a new insight into the role of KMT2D in migration and invasion of PTC.

MAP7 Promotes Breast Cancer Cell Migration and Invasion by Regulating the NF-B Pathway.

OBJECTIVE: To investigate and explore the molecular mechanisms of MAP7 on breast cancer cell migration and invasion. METHODS: The MAP7 transcript data in TCGA database was firstly statistically analyzed. Then, immunohistochemistry and western blot assays were applied to check MAP7 expression levels in breast cancer tissues or cell lines. EdU immunofluorescent staining assay was applied to reveal the cell proliferation of breast cancer cells after knockdown or overexpression of MAP7. Scratch and Transwell assays were applied to observe cell invasion and migration after knockdown or overexpression of MAP7. The western blot assays were employed to prove the expression levels of NF-B p65 and IBα after knockdown or overexpression of MAP7. Finally, breast xenograft model was established to verify the tumor volume and weight in mice. RESULTS: The results indicated the mRNA and protein expression of MAP7 was higher in breast cancer tissues or cell lines than that in normal tissue or normal breast epithelial cells, respectively. MAP7 promoted proliferation, migration, and invasion of breast cancer cells. Knockdown or overexpression of MAP7 in breast cancer cells would inhibit or promote phosphorylation of NF-B p65 and IBα protein. Finally, MAP7 can also promote tumor growth in mice. CONCLUSIONS: MAP7 facilitated breast cancer cell migration and invasion by regulating the NF-B pathway.

T Cell Factor 4 Is Involved in Papillary Thyroid Carcinoma via Regulating Long Non-Coding RNA HCP5.

The annual incidence of papillary thyroid carcinoma has increased dramatically. T cell factor 4 (TCF4) is an important component of Wnt signaling pathway.However, the role of TCF4 in PTC remains unknown. In this study, TCF4 was observed to overexpress in PTC patients and cells by qRT-PCR assay. The colony formation assay, Edu staining and transwell assay indicated thatoverexpression of TCF4 promoted cell proliferation and invasion of TCP-1 cells, whereas knockdown of TCF4 inhibited cell proliferation and invasion of IHH-4 cells. To investigate the mechanism of TCF4 in PTC cells, the luciferase assay demonstrated that TCF4 could modulate HCP5 expression. Besides, GLuc-ON promoter reporter assayproved that TCF4 could bind to HCP5 promoter. Further, knockdown of HCP5 could significantly up-regulated miR-15a, miR-216a-5p, miR-22-3p, miR-139-5p, miR-203, miR-27a-3p and miR-320, and down-regulated miR-186-5p in IHH-4 cells, which might be potential downstream of TFC4/HCP5 axis. In conclusion, up-regulation TCF4 can promote HCP5 expression via binding to HCP5 promoter. It may be the first time to prove that TCF4 regulates HCP5 in PTC, which provides a novel sight for treatment of PTC.

Impact of Nischarin on EMT regulators in breast cancer cell lines.

Nischarin is an integrin-binding protein, which is well known as a novel tumor suppressor. In breast cancer, Nischarin serves a critical role in breast cancer cell migration and invasion. However, the molecular mechanism underlying the role of Nischarin remains unclear. Recent findings have demonstrated that epithelial-mesenchymal transition (EMT) increases the capacity of cell migration and invasion. As a member of the integrin family, it was hypothesized that Nischarin may regulate cellular processes via various signaling pathways associated with the EMT process. The present study detected the mRNA levels of EMT regulators via reverse transcription-quantitative PCR and related protein levels via western blotting in breast cancer cells, following NISCH-overexpression and -knockdown. The results demonstrated that Nischarin inhibits cell proliferation, migration and invasion in breast cancer cells. Furthermore, when the NISCH gene was overexpressed, the relative mRNA level of E-cadherin was increased, while the relative mRNA levels of several transcription factors, such as Snail, ZEB1, N-cadherin, Slug, Twist1 and vimentin, decreased. When NISCH was silenced, these results were reversed. The present results demonstrated that Nischarin suppresses cell migration and invasion via inhibiting the EMT process.

The genomic mutation spectrums of breast fibroadenomas in Chinese population by whole exome sequencing analysis.

Fibroadenomas (FAs) are the most common fibroepithelial lesions and the most common benign tumors of the breast in women of reproductive age. Although MED12 mutations, an overwhelming majority of all mutations, and some other gene mutations have been found in FAs, the genomic landscapes of FAs are still not completely clear and the genomic mutation spectrums of FAs in Chinese population remains unknown. Here, by performing whole exome sequencing of 12 FAs and the corresponding normal breast tissues in Chinese Han population, we observed the somatic and germline landscapes of genetic alterations. We identified 16 recurrently mutated genes with 37 nonsynonymous or frameshift somatic mutations and 27 recurrent somatic copy number variants (CNVs). In these mutated genes, MED12 was the most common in FAs, harboring 6 nonsynonymous/frameshift somatic mutations and 1 CNV. In addition, 6 germline mutations of tumor susceptibility genes in 5 FAs were identified and the tumor mutational burden of the 5 FAs was significantly higher than the other 7 FAs without germline mutations. This study provides genomic mutation spectrums of FAs in Chinese population and expand the genetic spectrum of FAs.

Novel germline mutations and unclassified variants of BRCA1 and BRCA2 genes in Chinese women with familial breast/ovarian cancer.

BACKGROUND: Germline mutations in the BRCA1 and BRCA2 genes greatly increase a woman's risk of developing breast and/or ovarian cancer. The prevalence and distribution of such mutations differ across races/ethnicities. Several studies have investigated Chinese women with high-risk breast cancer, but the full spectrum of the mutations in these two genes remains unclear. METHODS: In this study, 133 unrelated Chinese women with familial breast/ovarian cancer living in Zhejiang, eastern China, were enrolled between the years 2008 and 2014. The complete coding regions and exon-intron boundaries of BRCA1 and BRCA2 were screened by PCR-sequencing assay. Haplotype analysis was performed to confirm BRCA1 and BRCA2 founder mutations. In silico predictions were performed to identify the non-synonymous amino acid changes that were likely to disrupt the functions of BRCA1 and BRCA2. RESULTS: A total of 23 deleterious mutations were detected in the two genes in 31 familial breast/ovarian cancer patients with a total mutation frequency of 23.3% (31/133). The highest frequency of 50.0% (8/16) was found in breast cancer patients with a history of ovarian cancer. The frequencies of BRCA1 and BRCA2 mutations were 13.5 % (18/133) and 9.8% (13/133), respectively. We identified five novel deleterious mutations (c.3295delC, c.3780_3781delAG, c.4063_4066delAATC, c.5161 > T and c.5173insA) in BRCA1 and seven (c.1-40delGA, c.4487delC, c.469_473delAAGTC, c.5495delC, c.6141T > A, c.6359C > G and c.7588C > T) in BRCA2, which accounted for 52.2% (12/23) of the total mutations. Six recurrent mutations were found, including four (c.3780_3781delAG, c.5154G > A, c.5468-1del8 and c.5470_5477del8) in BRCA1 and two (c.3109C > T and c.5682C > G) in BRCA2. Two recurrent BRCA1 mutations (c.5154G > A and c.5468-1del8) were identified as putative founder mutations. We also found 11 unclassified variants, and nine of these are novel. The possibility was that each of the non-synonymous amino acid changes would disrupt the function of BRCA1 and BRCA2 varied according to the different algorithms used. CONCLUSIONS: BRCA1 and BRCA2 mutations accounted for a considerable proportion of hereditary breast/ovarian cancer patients from eastern China and the spectrum of the mutations of these two genes exhibited some unique features. The two BRCA1 putative founder mutations may provide a cost-effective option to screen Chinese population, while founder effects of the two mutations should be investigated in a lager sample size of patients.

Drug response to HER2 gatekeeper T798M mutation in HER2-positive breast cancer.

The gatekeeper T798M mutation in HER2 kinase domain has been observed to considerably shift drug sensitivity to HER2 in breast cancer therapy. Here, drug response of clinical tyrosine kinase inhibitors (TKIs) to the mutation was profiled using a synthetic biology protocol. It was found that TKIs can be grouped into three classes in terms of their response behavior to T798M mutation: class I inhibitors exhibit drug resistance upon the mutation, such as lapatinib, TAK-285 and AEE788; class II inhibitors are insensitive to the mutation, such as erlotinib and gefitinib; and class III inhibitors can be sensitized by the mutation, such as staurosporine. However, kinetic study indicated that the mutation has only a modest effect on the binding of substrate ATP to HER2. Binding free energy analysis revealed that the drug response is primarily determined by direct interaction between the kinase and inhibitors, but not by indirect kinase interaction with competitive ATP. This is different to the molecular mechanism of "generic" drug resistance conferring from EGFR gatekeeper T790M mutation, which is caused by increased ATP affinity upon the mutation. Structural analysis of kinase-inhibitor complexes unraveled that HER2 T798M mutation induces significant steric hindrance to class I inhibitors, but can establish additional nonbonded interactions for class III inhibitors.

Expression of integrin-binding protein Nischarin in metastatic breast cancer.

The present study aimed to investigate the expression of Nischarin protein in primary breast cancer (PBC), and to evaluate its role in tumor metastasis. Paired specimens of breast cancer tissues and adjacent normal tissues were surgically obtained from 60 patients with PBC at the Zhejiang Cancer Hospital (Hangzhou, China). Nischarin protein concentrations were determined by an ELISA assay. Breast cancer tissues exhibited a significantly lower concentration of Nischarin (5.86 ± 3.19 ng/ml) compared with that of the adjacent noncancerous tissues (9.25 ± 3.65 ng/ml; P<0.001). Furthermore, cancer tissue from patients with lymph node metastasis had significantly lower levels of Nischarin protein (4.69 ± 2.40 ng/ml) than those of patients without lymph node metastasis (7.04 ± 3.47 ng/ml; P=0.004). There was no significant difference in Nischarin protein expression levels between patients with grade I, II or III PBC (grade I, 5.44 ± 3.57 ng/ml; grade II, 6.42 ± 3.85 ng/ml and grade III, 5.10 ± 1.18 ng/ml; P=0.765). The significant differences in the expression of Nischarin between: i) Cancer tissue and noncancerous tissue and ii) patients with and without lymph node metastasis, suggested that Nischarin may have a significant role in tumor occurrence and metastasis of breast cancer. Nischarin expression may therefore be used as a marker to predict the invasiveness and metastasis of PBC.

Serum tissue polypeptide-specific antigen is an independent predictor in breast cancer.

Tissue polypeptide-specific antigen (TPS) is a tumor proliferative marker associated with cytokeratin 18. The aim of the study was to investigate the potential relationship between the preoperative serum TPS levels and the outcome in Chinese breast cancer patients. 975 consecutive female patients, affected by invasive breast cancer under investigation from January 2005 to December 2011, had their TPS levels measured with a one-step solid phase radiometric sandwich assay detecting the M3 epitope on cytokeratin 18 fragments. The cut-off value was 80U/L. The average age diagnosed with breast cancer was 48, ranging from 23 to 71. About 19% (185) patients displayed an elevated preoperative TPS level (>80U/L) associated with older age (>45), advanced cancer stage, larger tumor size (>2cm), axillary lymph node metastasis, negative progesterone receptor status, and positive HER2 status. In addition, preoperative TPS levels were also significantly connected with recurrence (p<0.05), particularly distant metastasis and visceral metastasis. The mean preoperative TPS level was 68.4±116.43U/L (range 0-1839U/L). In multivariate analysis, high preoperative TPS level was recognized as an independent prognostic factor for disease-free survival (p<0.001 and overall survival (p=0.023). From these results we conclude that the serum preoperative TPS level may be a valuable and independent marker for breast cancer.

A support vector machine model for predicting non-sentinel lymph node status in patients with sentinel lymph node positive breast cancer.

This study aimed to investigate the accuracy and feasibility of support vector machine (SVM) modeling in predicting non-sentinel lymph node (NSLN) status in patients with SLN-positive breast cancer. Clinicopathological data were collected from 201 cases with sentinel lymph node biopsy breast cancer and included patient age, tumor size, histological type and grade, vascular invasion, estrogen receptor status, progesterone receptor status, CerbB2 status, size and number of positive SLNs, number of negative SLNs, and positive SLN membrane invasion. Feature vector selection was based on a combination of statistical filtration and model-dependent screening. The arbitrary combination with the smallest p value for SVM input was selected, the predicative results of the model were evaluated by a 10-fold cross validation, and a training model was established. Using SLN-positive patients as a double-blind test set, 85 patients were input into the model to analyze its sensitivity and specificity. The combination with the highest cross-validation accuracy was selected for the SVM model and consisted of the following: the number and size of positive SLNs, the number of negative SLNs, and the membrane invasion of positive SLNs. The training accuracy of the model established with the four variables was 92 %, and its cross-validation veracity was 87.6 %. The accuracy of an 85-patient double-blind test of the SVM model was 91.8 %. In conclusion, this SVM model is an accurate and feasible method for the prediction of NSLN status in SLN-positive breast cancer and is conducive to guide clinical treatment.

Low expression of stathmin in tumor predicts high response to neoadjuvant chemotherapy with docetaxel-containing regimens in locally advanced breast cancer.

AIMS: We performed this retrospective study to evaluate the value of clinicopathological factors and a novel molecular marker stathmin in predicting treatment response to neoadjuvant chemotherapy (NCT) with docetaxel-containing regimens in patients with locally advanced breast cancer. METHODS: Fifty-four consecutive locally advanced patients receiving docetaxel-containing NCT between January 2006 and July 2010 in Zhejiang Cancer Hospital were included. The expression levels of estrogen receptor (ER), progesterone receptor (PgR), epidermal growth factor receptor-2 (HER-2), and p53 were detected by immunohistochemistry, while expression of stathmin mRNA was measured by Quanti-Gene assay. RESULTS: The overall clinical objective response (cOR) rate was 75.9% (41/54) in breast. A total of 34 patients (63.0%) experienced pathological OR (pOR), with pathological complete remission (pCR) rate of 20.4% (11/54) in breast and 16.7% (9/54) in both breast and axilla. In univariate analysis, there were associations of pOR in both breast and axilla with age (p=0.054), ER status (p=0.059), subtypes (p=0.062), p53 (p=0.030), and stathmin expression (three terciles) (p=0.039). Mean expression of stathmin in pOR group was 0.410, compared with that in no response group of 0.556 (p=0.051 by Student's t-test). Similarly, a lower expression of stathmin might represent a higher pCR rate (p=0.061). Moreover, the LOWESS smoothing plot showed the same trend, that is, that tumor with a lower level of stathmin expression had a higher probability of response to docetaxel-containing NCT. After multivariate adjustment, both ER and stathmin remained significant with hazard ratio of 4.58 (95% CI: 1.11-18.94, p=0.036) and 2.94 (95% CI: 1.26-6.86, p=0.012), respectively. CONCLUSIONS: In conclusion, ER and stathmin were independent predictive factors for NCT with docetaxel-containing regimens.

Unilateral hemilaminectomy for patients with intradural extramedullary tumors.

A modified hemilaminectomy was introduced in an attempt to explore the operative techniques and the values of the limited approach to spinal cord tumors. Forty-five consecutive patients with intradural extramedullary lesions, who underwent modified hemilaminectomy, were studied retrospectively. The intraspinal tumors were removed via the limited bone window with a 3.3-cm mean length (range: 2.0-6.5 cm) and a 1.2-cm mean width (range: 0.6-1.5 cm), in which the inner parts of the medial and lateral laminae were mostly undercut for wider view. Spinal lesions were cervical in 21 cases, thoracic in 12 cases, lumbar in 10 cases, and multiple in 2 cases. Forty-three cases were completely excised via hemilaminectomy alone. Two subjects with dumbbell neurinoma underwent two-stage tumor removal via anterolateral cervical approach following hemilaminectomy. With respect to neurological status, the percentage of good Frankel scale (D+E grade) was markedly improved from 22.2% on admission to 93.3% at follow-up. At the median 26-month follow-up evaluation by magnetic resonance imaging (MRI), none of the subjects showed spinal deformity or instability. By preserving musculoligamentous attachments and posterior bony elements as much as possible, the modified approach is minimally invasive and may be routinely used to remove intradural and extramedullary tumors, especially in patients with meningiomas and neurinomas.