CRABP2 involvement in a mechanism of Golgi stress and tumor dry matter in non-small cell lung cancer cells via ER dependent Hippo pathway

Objective: The paper aimed to explore the mechanism of cellular retinoic acid binding protein 2 (CRABP2) involvement in Golgi stress and tumor dryness in non-small cell lung cancer (NSCLC) cells through the estrogen receptor (ER) dependent Hippo pathway. Methods: Human NSCLC cell line A549 was purchased from ATCC andcultured in RPMI-1640 with 10% FBS. Attractene reagent was used for plasmid transfection. ER (sh) RNA was designed using RNAi Designer. Seventy-six hours after infection, stable cells were obtained after treated with puromycin for 3 weeks. ER silencing cells (with inhibited ER expression) were compared to the control cells (normal cultured NSCLC cell line A549, CRABP2 normal expression). CRABP2 and ER expression levels were detected by RT-PCR. MTT assay was used to detect cell proliferation, and the cell localization of ER and Golgi was observed by confocal microscopy. The invasion and metastasis of cells were analyzed by Boden chamber invasion and migration assays. Western blotting assays was used for detecting the protein expression of E-cadherin, vimentin, ZO-1 protein and epithelial-mesenchymal transition (EMT) related factors. Results: The lower expression level of mRNA was detected in the ER-silencing group compared to the control group (P<0.05). We also found a higher proliferation level of cells, the number of invading and metastatic cells, the expression of vimentin, p-Lats1T1079, Lats1 and p-YAPS127 mRNA in the control group compared to the ER silencing group (P<0.05). And the expression level of protein kinase RNA-like endoplasmic reticulum kinase (PERK), phosphorylate eukaryotic initiation factor 2 (p-eIF2 alpha), activating transcription factor 4 (ATF4) and C/EBP-homologous protein (CHOP) in the control group was higher than that in the ER silencing group (P<0.05). Adversely, a lower expression level of E-cadherin and ZO-1 protein was found in the control group compared to the ER silencing group (P<0.05). Conclusion: The expression of CRABP2 in NSCLC cells was regulated by ER, and cell proliferation and invasion were regulated by the Hippo pathway. At the same time, it was found that decreased expression of CRABP2 enhanced endoplasmic reticulum/Golgi stress response.

Correlation of breast cancer microcirculation construction with tumor stem cells (CSCs) and epithelial-mesenchymal transition (EMT) based on contrast-enhanced ultrasound (CEUS)

Objective This study is to explore the correlation between the contrast-enhanced ultrasound (CEUS) characteristics of breast cancer and the epithelial-mesenchyme transformation (EMT). Methods Totally 119 patients of breast cancer underwent CEUS. Tissues in the active area were collected and subjected to the immunohistochemical detection, PT-PCR and Western blot. Correlation analysis was conducted between the clinical pathological parameters and the CEUS indicators. Results The expression levels of CD44, N-cadherin, and β-catenin in breast cancer tissues were higher than those in adjacent tissues (P<0.05). However, the expression levels of CD24 and E-cadherin in breast cancer tissues were lower than those in adjacent tissues (P<0.05). There was no significant difference in E-cadherin mRNA and Vimentin levels between cancer and adjacent tissues (P>0.05). The expressions were up-regulated in the CSCs, with higher histological grade, lymph node metastasis, and negative estrogen receptor (ER) expression. Smaller breast tumors, with no lymph node metastasis, lower clinical stage, and positive ER expression, tended to exhibit the up-regulated epithelial phenotype. Breast tumors, with high histological grade, lymph node metastasis, high clinical staging grade, and negative ER expression, tended to exhibit the up-regulated interstitial phenotype. The peak intensity of the time-intensity curve (TIC) for the CEUS was positively correlated with the CSC marker CD44 and the interstitial phenotype marker N-cadherin. The starting time of enhancement was negatively correlated with the N-cadherin. Area under the curve was positively correlated with the expression of CD44 and N-cadherin, while negatively correlated with the epithelial phenotype marker β-catenin. The time to peak was negatively correlated with the interstitial phenotypes Vimentin and N-cadherin, with no correlation with the E-cadherin or β-catenin. Conclusion Breast cancers show the enlarged lesions after enlargement and perfusion defect for the CEUS. The fast-in pattern, high enhancement, and high perfusion in the TIC are correlated with the CSCs and EMT expressions, suggesting poor disease prognosis.

Androgen Receptor Expression Associates With Distinctive Clinicopathological and Molecular Features in ER-Positive and ER-Negative Breast Cancer

Background: Androgen receptor (AR) expression is frequently observed in breast cancer, but its association with estrogen receptor (ER) expression of breast cancer remains unclear. Methods: In this study, we analyzed the clinicopathological and molecular features associated AR loss in ER-positive and ER-negative breast cancer respectively, trying to elucidate the molecular correlation between AR and ER. Results: Our results showed that AR loss was associated with different clinicopathological characteristics in ER-positive and ER-negative breast cancer. Moreover, the expression of AR was correlated with different molecular features in ER-positive and ER-negative breast cancer.Conclusions: These results suggest that the role of AR in ER-positive breast cancer is distinctive from that in ER-negative breast cancer.

Analyzing the Estrogen Receptor Status of Liver Metastases with [18F]-FES-PET in Patients with Breast Cancer

Background: Positron emission tomography (PET) with 16α-[18F]-fluoro-17β-estradiol ([18F]-FES) can visualize estrogen receptor (ER) expression, but it is challenging to determine the ER status of liver metastases, due to high physiological [18F]-FES uptake. We evaluated whether [18F]-FES-PET can be used to determine the ER status of liver metastases, using corresponding liver biopsies as the gold standard. Methods: Patients with metastatic breast cancer (n = 23) were included if they had undergone a [18F]-FES-PET, liver metastasis biopsy, CT-scan, and [18F]-FDG-PET. [18F]-FES-PET scans were assessed by visual and quantitative analysis, tracer uptake was correlated with ER expression measured by immunohistochemical staining and the effects of region-of-interest size and background correction were determined. Results: Visual analysis allowed ER assessment of liver metastases with 100% specificity and 18% sensitivity. Quantitative analysis improved the sensitivity. Reduction of the region-of-interest size did not further improve the results, but background correction improved ER assessment, resulting in 83% specificity and 77% sensitivity. Using separate thresholds for ER+ and ER– metastases, positive and negative predictive values of 100% and 75%, respectively, could be obtained, although 30% of metastases remained inconclusive. Conclusion: In the majority of liver metastases, ER status can be determined with [18F]-FES-PET if background correction and separate thresholds are applied.

Androgen Receptor Expression Associates With Distinctive Clinicopathological and Molecular Features in ER-Positive and ER-Negative Breast Cancer

Background Androgen receptor (AR) expression is frequently observed in breast cancer, but its association with estrogen receptor (ER) expression of breast cancer remains unclear. Methods In this study, we analyzed the clinicopathological and molecular features associated AR loss in ER-positive and ER-negative breast cancer respectively, trying to elucidate the molecular correlation between AR and ER. Results Our results showed that AR loss was associated with different clinicopathological characteristics in ER-positive and ER-negative breast cancer. Moreover, the expression of AR was correlated with different molecular features in ER-positive and ER-negative breast cancer. Conclusions These results suggest that the role of AR in ER-positive breast cancer is distinctive from that in ER-negative breast cancer.

Impact of Estrogen Receptor Expression on Prognosis of Ovarian Cancer According to Antibody Clone Used for Immunohistochemistry: A Meta-analysis

BackgroundThe prognostic value of the expression of estrogen receptor (ER) subtypes ER⍺ and ERβ in ovarian cancer has previously been evaluated by meta-analyses. However, the results are contradictory and controversial. MethodsWe conducted an updated meta-analysis with stringent inclusion criteria to ensure homogeneous studies to determine the effect of ER subtypes on ovarian cancer prognosis. Articles were retrieved by systematic search of PubMed and Web of Science for articles dated up to June 2021. Only studies with known hazard ratio (HR) and antibody clone for immunochemistry (IHC) were included. Pooled HRs with the corresponding 95% confidence intervals (CIs) were calculated for the effect of ER⍺ and ERβ expression on ovarian cancer patient progression-free survival (PFS) and overall survival (OS).ResultsA total of 17 studies were included, of which 11 and 13 studies examined the relationships between ER⍺ expression and PFS and OS, respectively, and 5 and 7 studies examined the relationships between ERβ expression and PFS and OS, respectively. Neither ER⍺ expression (random-effects model; HR=0.99, 95% CI=0.83-1.18) nor ERβ expression (fixed-effects model; HR=0.94, 95% CI=0.69-1.27) was associated with PFS. Random-effects models showed that ER⍺ expression (HR=0.81, 95% CI=0.64-1.02) and ERβ expression (HR=0.75, 95% CI=0.50-1.13) were only marginally and not significantly associated with better OS. Subgroup analysis revealed that ER⍺ expression determined using antibody clone 1D5 (HR=0.75, 95% CI=0.64-0.88) and ERβ expression determined using ERβ1-specific-antibody clone PPG5/10 or EMR02 (HR=0.65, 95% CI=0.50-0.86) were associated with significantly better OS, but ER expression determined using other antibodies was not.ConclusionsBoth ER⍺ expression and ERβ expression determined using certain antibody clones are significantly associated with OS of ovarian cancer patients, which suggests that both ER subtypes are prognostic biomarkers for ovarian cancer. The findings of this study provide new insight into the impact of ER expression on ovarian cancer prognosis.

A Real-World-Data Retrospective Cohort Study of Low Estrogen Receptor Positive Early Breast Cancer: Natural History and Treatment Outcomes.

Purpose Estrogen receptor–positive (ER+) breast cancer (BC) is a heterogeneous disease with an ongoing debate regarding the optimal cutoff point for clinically relevant ER expression. We used a real-world database to assess prognostic and predictive values of lower ER expression levels on treatment outcomes with endocrine therapy. Methods We used a nationwide electronic health record– (EHR-) derived deidentified database. Descriptive statistics were used to evaluate the correlation between ER expression, tumor characteristics, and treatment patterns among patients with early-stage BC. We used Kaplan-Meier survival curves to estimate relapse-free survival (RFS) and overall survival (OS). To assess an optimal ER expression level cut point, correlations between ER+ expression and clinical outcomes were performed. Results Among 4697 patients with early-stage BC, 83 (1.76%) had ER+-low BC (ER expression, 1%-9.99%), and 36 (0.8%) had ER+-intermediate BC (10%-19.9%). ER+-low tumors were associated with higher tumor grade, larger size and higher axillary tumor burden than ER+-high tumors (≥ 20% ER expression). African Americans had a higher prevalence of both triple-negative BC (TNBC; 21%) and ER+-low BC (22%) than ER+-high tumors (8%). Patients with ER+-low and ER+-intermediate tumors had survival outcomes similar to patients with TNBC and worse survival outcomes than patients with ER+-high tumors (P < .001). No significant correlations between endocrine therapy and RFS or OS were observed among patients with either ER+-low or ER+-intermediate BCs. Further tumors with < 20% ER expression were associated with worse outcomes. Conclusions Patients with ER expression levels < 20% derive minimal benefit from endocrine therapies.

Effect of Estrogen Receptor Expression Level and Hormonal Therapy on Prognosis of Early Breast Cancer

PurposeEstrogen receptor (ER) expression in breast cancer plays an essential role in carcinogenesis and disease progression. Recently, tumors with low level (1-10%) of ER expression have been separately defined as ER Low Positive (ERlow). It is suggested that ERlow tumors might be morphologically and behaviorally different from tumors with high ER expression (ERhigh).MethodsRetrospective analysis of a prospective cohort database was performed. Patients who underwent curative surgery for early breast cancer and had available medical records were included for analysis. Difference in clinicopathological characteristics, endocrine responsiveness and five-year recurrence-free survival was evaluated between different ER subgroups (ERhigh, ERlow, and ER-negative (ER-)).ResultsA total of 2162 breast cancer patients were included in the analysis, Tis and T1 stage. Among them, 1654 (76.5%) were ERhigh, 54 (2.5%) were ERlow, and 454 (21.0%) were ER- patients. ERlow cases were associated with smaller size, higher histologic grade, positive human epidermal growth factor receptor 2 (HER2), negative progesterone receptor, and higher Ki-67 expression. Recurrence rate was highest in ER- tumors and was inversely proportional to ER expression. Recurrence-free survival was not affected by hormonal therapy in the ERlow group (P = 0.418).ConclusionERlow breast cancer showed distinct clinicopathological features. ERlow tumors seemed to have higher recurrence rates compared to ERhigh tumors, and they showed no significant benefit from hormonal therapy. Future large scale prospective studies are necessary to validate the treatment options for ERlow breast cancer.

[18F]-Fluoroestradiol PET/CT: a modern look at nuclear medicine applications

Breast cancer is one of the most commonly diagnosed cancers and the leading cause of cancer mortality among women. Approximately 70–80 % of breast cancers are estrogen (ER) and/or progesterone receptor-positive, thus making endocrine therapy an important stage of treatment. Receptor expression in breast cancer cells is usually assessed by tissue immunohistochemistry. The method of positron emission tomography, combined with computed tomography (PET/CT), makes it possible to evaluate not only anatomical and structural, but also metabolic changes in tumor tissue. 18F-Fluoroestradiol (18F-FES) is a radiopharmaceutical drug, an estradiol analogue, which is used in the diagnostics of ER-expressing tumors and is utilized for detection and quantification of ER expression in vivo. Various studies show that 18F-FES accumulation indicates presence of ER-positive tumor tissue, which, in most cases, is confirmed by tissue immunohistochemistry. Although current guidelines recommend 18F-fluorodeoxyglucose PET/CT when routine examinations demonstrate ambiguous results, 18F-FES PET/CT can be the preferable imaging modality in the diagnostics of ER-positive breast cancer. It should be noted, that PET/CT with 18F-FES can also be effective for evaluation of tumors with a high level of ER expression, like ovarian cancer.