ATRX is a predictive marker for endocrinotherapy and chemotherapy resistance in HER2-/HR+ breast cancer through the regulation of the AR, GLI3 and GATA2 transcriptional network.

Drug resistance in breast cancer (BC) is a clinical challenge. Exploring the mechanism and identifying a precise predictive biomarker for the drug resistance in BC is critical. Three first-line drug (paclitaxel, doxorubicin and tamoxifen) resistance datasets in BC from GEO were merged to obtain 1,461 differentially expressed genes for weighted correlation network analysis, resulting in identifying ATRX as the hub gene. ATRX is a chromatin remodelling protein, therefore, ATRX-associated transcription factors were explored, thereby identifying the network of AR, GLI3 and GATA2. GO and KEGG analyses revealed immunity, transcriptional regulation and endocrinotherapy/chemotherapy resistance were enriched. Moreover, CIBERSORT revealed immunity regulation was inhibited in the resistance group. ssGSEA showed a significantly lower immune status in the ATRX-Low group compared to the ATRX-High group. Furthermore, the peaks of H3K9me3 ChIP-seq on the four genes were higher in normal tissues than in BC tissues. Notably, the frequency of ATRX mutation was higher than BRCA in BC. Moreover, depressed ATRX revealed worse overall survival and disease-free survival in the human epidermal growth factor receptor 2 (HER2)-/hormone receptor (HR)+ BC. Additionally, depressed ATRX predicted poor results for patients who underwent endocrinotherapy or chemotherapy in the HER2-/HR+ BC subgroup. A nomogram based on ATRX, TILs and ER exhibited a significantly accurate survival prediction ability. Importantly, overexpression of ATRX significantly inhibited the IC50 of the three first-line drugs on MCF-7 cell. Thus, ATRX is an efficient predictive biomarker for endocrinotherapy and chemotherapy resistance in HER2-/HR+ BC and acts by suppressing the AR, GLI3 and GATA2 transcriptional network.

A Preliminary Study of Ultrasound-Guided Microwave Ablation for Nonpuerperal Mastitis Treatment.

Introduction: This study investigated the feasibility of ultrasound (US)-guided microwave ablation (MWA) as a treatment for nonpuerperal mastitis (NPM). Methods: Fifty-three patients with NPM diagnosed by biopsy and treated with US-guided MWA at the Affiliated Hospital of Nantong University between September 2020 and February 2022 were classified according to whether they underwent MWA alone (n = 29) or MWA with incision and drainage (n = 24). Patients were followed up by interviews, physical and US examinations, and evaluation of breast skin at 1 week and 1, 2, and 3 months after treatment. Data from these patients were prospectively collected and retrospectively analyzed. Results: The overall mean patient age was 34.42 ± 9.20 years. The groups differed significantly by age, involved quadrants, and the initial maximum diameter of lesions. In the MWA group, the cure rate was 34.48%, and the apparent efficiency rate was 65.52%. In the MWA with incision and drainage, the apparent efficiency rate was 91.66%, and the effective rate was 4.17%. The excellent rate for breast aesthetics in the MWA group was 79.31%, and the good rate was 20.69%. The excellent rate in the MWA with incision and drainage group was 45.83%, the good rate was 41.67%, and the qualified rate was 12.5%. The mean maximum diameter of lesions in the two groups decreased significantly. Conclusion: For NPM with small lesions in a single quadrant, MWA therapy is a direct and effective method. For larger lesions involving two or more quadrants, the combined treatment of MWA with incision and drainage showed significant improvement in a short period. MWA treatment of NPM has importance for further research and clinical applications.

Lanostane triterpenoids from the fruiting bodies of Ganoderma hainanense and their cytotoxic activity.

Three undescribed lanostane triterpenoids, 24E-en-11-oxo-ganoderiol D (1), 11ß-hydroxy-ganoderiol D (2), and 11ß-hydroxy-lucidone H (3) were isolated from the 80% EtOH extract of the fruiting bodies of Ganoderma hainanense. Structural elucidation of all the compounds were performed by spectral methods such as 1 D and 2 D (1H-1H COSY, HMQC, and HMBC) NMR spectroscopy. All the triterpenoids were in vitro evaluated for their cytotoxic activities against six mammary adenocarcinoma cell lines (MCF7, MDA-MB-231, SK-BR-3, BT-20, HCC38, and AU565). As a result, compound 3 exhibited significant cytotoxic activities against all tested cell lines with IC50 values less than 20 µM.

Knockdown of Ran GTPase expression inhibits the proliferation and migration of breast cancer cells.

Breast cancer is the second leading cause of cancer­associated mortality in women worldwide. Strong evidence has suggested that Ran, which is a small GTP binding protein involved in the transport of RNA and protein across the nucleus, may be a key cellular protein involved in the metastatic progression of cancer. The present study investigated Ran gene expression in breast cancer tissue samples obtained from 140 patients who had undergone surgical resection for breast cancer. Western blot analysis of Ran in breast cancer tissues and paired adjacent normal tissues showed that expression of Ran was significantly increased in breast cancer tissues. Immunohistochemistry analyses conducted on formalin­fixed paraffin­embedded breast cancer tissue sections revealed that Ran expression was associated with tumor histological grade, nerve invasion and metastasis, vascular metastasis and Ki­67 expression (a marker of cell proliferation). Kaplan­Meier survival analysis showed that increased Ran expression in patients with breast cancer was positively associated with a poor survival prognosis. Furthermore, in vitro experiments demonstrated that highly migratory MDA­MB­231 cancer cells treated with Ran­si­RNA (si­Ran), which knocked down expression of Ran, exhibited decreased motility in trans­well migration and wound healing assays. Cell cycle analysis of Ran knocked down MDA­MB­231 cells implicated Ran in cell cycle arrest and the inhibition of proliferation. Furthermore, a starvation and re­feeding (CCK­8) assay was performed, which indicated that Ran regulated breast cancer cell proliferation. Taken together, the results provide strong in vitro evidence of the involvement of Ran in the progression of breast cancer and suggest that it could have high potential as a therapeutic target and/or marker of disease.

Suppression of Kpnß1 expression inhibits human breast cancer cell proliferation by abrogating nuclear transport of Her2.

Breast cancer (BC) is one of the most fatal diseases and poses critical health problems worldwide. However, its mechanisms remain unclear. Consequently, there is an urgency to investigate the mechanisms involved in BC initiation and progression and identify novel therapeutics for its prevention and treatment. In this study, we identified karyopherin ß-1 (Kpnß1) as a possible novel therapeutic target for BC. Western blotting was used to evaluate the expression of Kpnß1 in four pairs of tumorous and adjacent non-tumorous tissues. The results revealed that the protein level of Kpnß1 was higher in the cancer samples compared with those in the corresponding normal samples. Immunohistochemistry was performed on 140 BC cases and indicated that Kpnß1 was significantly associated with clinical pathological variables. Kaplan-Meier curve revealed that high expression of Kpnß1 was related to poor BC patient prognosis. A starvation and re-feeding assay was used to imitate the cell cycle using the SKBR-3 cell line, indicating that Kpnß1 plays a critical role in cell proliferation. The Cell Counting Kit-8 assay revealed that SKBR-3 cells treated with Kpnß1-siRNA (siKpnß1) grew more slowly than the control cells, while flow cytometry revealed that low-Kpnß1 expressing SKBR-3 cells exhibited increased BC cell apoptosis. Furthermore, the interaction between Kpnß1 and Her2 was clearly observed by immunoprecipitation, indicating that Kpnß1-knockdown abrogated nuclear transport of Her2. In summary, our findings revealed that Kpnß1 is involved in the progression of BC and may be a useful therapeutic target.

Expression of HAX1 and Ki-67 in breast cancer and its correlations with patient's clinicopathological characteristics and prognosis.

OBJECTIVE: This study aimed to investigate the HS-1-associated protein X-1 (HAX1) and Ki-67 expression in the breast cancer and its clinical significance. METHODS: Breast cancer tissues and tumor-adjacent tissues were collected from 81 patients, and immunohistochemistry was conducted to detect the HAX1 and Ki-67 expression. The correlations of HAX1 expression with demographics, clinicopathological characteristics and prognosis were evaluated. RESULTS: HAX1 was highly expressed in the breast cancer, and its expression was related to the degree of breast cancer differentiation (P=0.002), lymph nodes metastasis (P=0.008) and progesterone receptor (PR) (P=0.021), but not with the age, tumor size, histological type, estrogen receptor (ER), HER2 and p53. HAX1 expression was positively related to Ki-67 expression in the breast cancer (r(2)=0.394, P=0.019). In addition, a higher HAX-1 expression was related to a lower 10-year survival rate. CONCLUSION: HAX1 is a new protein related to the breast cancer and probably plays an important role in the invasion and metastasis of breast cancer. Thus, HAX1 may be used as a potential target for the therapy of breast cancer.

[Expression and clinicopathological significance of Emi1 in breast carcinoma].

OBJECTIVE: To examine the expression and prognostic value of novel oncogene Emi1 in breast cancer. METHODS: Immunohistochemical analysis was performed for 93 human breast carcinoma samples and adjacent normal tissues for detecting the expression of Emi1 and cell proliferative factor Ki-67. Then the data were correlated with clinicopathological features. RESULTS: Emi1 was highly expressed in breast carcinoma tissues compared with adjacent normal tissues. Emi1 expression was significantly associated with histological grade (P = 0.002) , axillary lymph node status (P = 0.017) and ER status (P = 0.038) . However, there was no correlation between Emi1 expression and other factors such as age, tumor size, histology, PR status, HER-2 or P53. Meanwhile similar results were obtained for Ki-67 expression. A marked correlation also existed between Emi1 and Ki-67 expression (Spearman's r² = 0.454, P = 0.002) . CONCLUSION: As a novel breast cancer associated gene, Emil plays an important role in evaluating invasion and metastatic potentiality in breast cancer. And it is also an important prognostic predictor of breast cancer so as to become a potential therapeutic target for breast cancer.

Decreased expression and prognostic role of cytoplasmic BRSK1 in human breast carcinoma: correlation with Jab1 stability and PI3K/Akt pathway.

OBJECTIVE: Jun activation domain-binding protein 1 (Jab1) was overexpressed in breast cancer, which was involved in degradation of the cyclin-dependent kinase inhibitor p27(Kip1). The objective of this study was to examine the effect of brain specific kinase 1 (BRSK1) expression on Jab1 over-expression and related signaling pathway in breast cancer. METHODS: Immunohistochemical analysis was performed in 95 human breast carcinoma samples and the data were correlated with clinicopathologic features. Furthermore, Western blot analysis was performed for BRSK1 and Jab1 in breast carcinoma samples and cell lines to evaluate their protein levels and molecular interaction. RESULTS: We found that the cytoplasmic BRSK1 expression was inversely associated with Jab1 expression (P<0.01) and correlated significantly with histologic grade (P=0.006), however nuclear BRSK1 expression couldn't obtain similar results. Kaplan-Meier analysis revealed that survival curves of low versus high expressers of cytoplasmic BRSK1 and Jab1 showed a highly significant separation in breast cancer (P<0.01). While in vitro, following release of breast cancer cell lines from serum starvation, the expression of Jab1, phosphor-Akt (p-Akt) was up-regulated, whereas BRSK1 and p27(Kip1) were decreased. Treatment of phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 could diminish Jab1 expression but increase BRSK1 expression. In addition, we employed siRNA technique to knock down Jab1 and/or BRSK1 expression and observed their effects on MDA-MB-231 cell growth. CONCLUSIONS: BRSK1 is a novel tumor suppressor in breast cancer which inversely correlated with Jab1 expression, may involve in the restoring Jab1-induced suppression of p27(Kip1) and may regulate cell cycle through the PI3K/Akt pathway.

Expression and prognostic role of SKIP in human breast carcinoma.

Ski-interacting protein (SKIP) is a nuclear hormone receptor-interacting cofactor, interactions with the proto-oncogene Ski, appears to modulate a number of signalling pathways involved in control of cell proliferation and differentiation, and may play a critical role in oncogenesis. In the present study, to investigate the potential roles of SKIP in breast cancer, expression patterns, interaction and the correlation with clinical/prognostic factors of SKIP and Ki-67 were examined among patients with breast cancer. Immunohistochemistry and Western blot analysis were performed for SKIP in 85 breast carcinoma samples. The data were correlated with clinicopathological features. The univariate and multivariate survival analyses were also performed to determine their prognostic significance. We found that SKIP was over expressed in breast carcinoma as compared with the adjacent normal tissues. High expression of SKIP was positively associated with histological grade (P = 0.01) and Ki-67 (P = 0.004). Univariate analysis showed that SKIP expression was associated with a poor prognosis (P = 0.006). While in vitro, following release of breast cancer cell lines from serum starvation, the expression of SKIP was up-regulated, whereas p27 was down-regulated. In addition, we employed small interfering RNA (siRNA) technique to knock down SKIP expression and observed it effects on MDA-MB-231 cells growth. SKIP depletion by siRNA inhibited cell proliferation, blocked S phase and decreased cyclin A and cyclin B levels. On the basis of these results, we suggested that SKIP overexpression was involved in the pathogenesis of breast cancer, which might serve as a future target for breast cancer.

The expression and prognosis of Emi1 and Skp2 in breast carcinoma: associated with PI3K/Akt pathway and cell proliferation.

S-phase kinase protein 2 (Skp2) is oncogenic and overexpressed in human breast cancer. The objective of this study was to examine the effect of early mitotic inhibitor-1 (Emi1) over-expression on Skp2 expression and related signaling pathway in breast cancer. Immunohistochemical analysis was performed in 98 human breast carcinoma samples and the data were correlated with clinicopathologic features. Furthermore, Western blot analysis was performed for Emi1 and Skp2 in breast carcinoma samples and cell lines to evaluate their protein levels and molecular interaction. We found that the expression of Emi1 was positively related with Skp2 expression (P < 0.01) and Emi1 expression correlated significantly with histologic grade (P = 0.005), meanwhile Skp2 expression obtained similar results. Kaplan-Meier analysis revealed that survival curves of low versus high expressers of Emi1 and Skp2 showed a highly significant separation in human breast cancer (P < 0.01). While in vitro, following release of breast cancer cell lines from serum starvation, the expression of Emi1, Skp2, phosphor-Akt (p-Akt) was up-regulated, whereas p27(Kip1) was down-regulated. Treatment of phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 could arrest cells growth and diminish Emi1 expression. These results suggested that Emi1's anti-apoptotic and proliferative abilities appear to be triggered at least in part by the modulation of Skp2, combined Emi1 and Skp2 expressions, may be prognostic for patients with invasive breast carcinomas, which also associated with p-Akt and enabled p27(kip1) degradation. Emi1 may serve as a potential therapeutic strategy aimed at PI3K for the management of breast cancer.

[Expression and prognostic value of galectin-9 in hepatocellular carcinoma patients].

OBJECTIVE: To explore the prognostic value of galectin-9 in patients with hepatocellular carcinoma (HCC). METHODS: Galectin-9 was validated by immunohistochemisty in tissue microarrays from HCC patients (n = 147) and statistically assessed for the prognosis. The serum levels of galectin-9 from another independent cohort including HCC patients (n = 31) were assessed by enzyme-linked immunosorbent assay (ELISA). RESULTS: Patients with a lower expression of galectin-9 had significantly worse prognosis than those with a higher expression. The serum level of galectin-9 of HCC patients (8.36 ± 2.12) µg/L was significantly lower than that in healthy (4.62 ± 1.59 )µg/L and liver cirrhosis controls (5.11 ± 1.92 )µg/L (P < 0.05). Multivariate Cox proportional hazard analysis showed that galectin-9 was an independent marker for predicting a poor prognosis of HCC patients. CONCLUSION: Involved in the poor prognosis of HCC patients, galectin-9 may be a new predictor of recurrence for HCC patients and serve as a high-priority therapeutic target.

Association of microRNA-196a-2 gene polymorphism with gastric cancer risk in a Chinese population.

BACKGROUND: It has been proposed that single nucleotide polymorphisms (SNPs) in microRNAs (miRNAs) could affect the expression of the miRNA and contribute to the susceptibility of human tumors. However, the role of genetic variant (T/C) in miR-196a-2 in gastric cancer susceptibility is still unknown. OBJECTIVES: To evaluate the association between genetic polymorphism of miR-196a-2 (rs11614913) and risk of gastric cancer, a hospital-based case-control study was conducted in a Chinese population. METHODS: The miR-196a-2 polymorphism was determined using the method of polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) in 213 gastric cancer patients and 213 age- and sex-matched controls. RESULTS: In the present study, we found that a significantly increased risk of gastric cancer in subjects with the variant homozygote CC of miR-196a-2 compared with wild-type homozygote TT and heterozygote CT carriers (adjusted odds ratio (OR) = 1.57, 95% confidence interval (CI) = 1.03-2.39, P = 0.038). Stratified analyses indicated that the variant homozygote CC genotype had a strong association with lymph node metastasis of gastric cancer (adjusted OR = 2.25, 95% CI = 1.21-4.18, P = 0.011). CONCLUSIONS: These findings suggest that the genetic variant within miR-196a-2 could play an important role in the development and progression of gastric cancer. We expect the findings may be helpful to better understand the mechanism of gastric carcinogenesis.