G protein subunit gamma 5 promotes the proliferation, metastasis and glycolysis of breast cancer cells through the Wnt/ß-catenin pathway.

GNG5 is suggested to exert a critical effect on tumor development in human beings; however, its function and related mechanism within breast cancer (BC) are still unclear. In this regard, the present work focused on identifying and evaluating GNG5's function and revealing its possible molecular mechanism. Subcutaneous tumorigenesis model of nude mice and in-vitro cell model was established. The relationship between GNG5 expression and BC was studied through knockdown and overexpression experiments. The proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of liver cancer cell lines overexpressing or silencing GNG5 were detected. Furthermore, the pathway mechanism of GNG5 was evaluated at the molecular level and was performed to further verify the possible targets and mechanisms of action. In comparison with that in normal tissue, GNG5 level within BC tissue was higher. In addition, GNG5 overexpression stimulated BC cell proliferation, invasion, migration and EMT. BC cells with reduced GNG5 expression exhibited significant decreases in glucose uptake, lactate levels, and ATP concentrations. In addition, GNG5 knockdown inhibited Wnt/ß-catenin signaling. This study indicates that GNG5 may generate a vital function in BC. The results of the current work demonstrated GNG5's effect on BC pathological process, also providing a reference for developing new targeted therapies for BC.

miR-26b-5p suppresses chemoresistance in breast cancer by targeting serglycin.

Chemoresistance is a crucial barrier to limit the therapeutic outcome of breast cancer (BC), and the mechanism underlying chemoresistance development in BC is not fully understood. In this study, we aimed to investigate the potential involvement of miR-26b-5p/serglycin (SRGN) axis in BC drug resistance. The expression level of SRGN in drug-resistant BC cells was investigated by western blotting analysis, real-time quantitative PCR (qRT-PCR), immunohistochemical staining, and ELISA. Its expression between chemoresistant and sensitive patient samples was compared by qRT-PCR. Bioinformatics tool and dual-luciferase reporter assay were employed to identify miR-26b-5p as a regulator of SRGN. Functional assays were performed to examine cell proliferation, cell viability, apoptosis, migration, and invasion ability in vitro. Xenograft tumorigenesis experiment was conducted to evaluate the tumor suppressor effect of miR-26b-5p on chemoresistant BC cells. SRGN expression was significantly upregulated in both chemoresistant BC cell lines and chemoresistant patient samples. miR-26b-5p was identified as an upstream regulator of SRGN. Overexpression of miR-26b-5p downregulated SRGN expression, overcame chemoresistance, and suppressed cell proliferation, migration, and invasion in BC cells. Overexpression of miR-26b-5p also suppressed the tumorigenesis of chemoresistant BC cells in vivo. Mechanistically, the downregulation of SRGN by miR-26b-5p decreased the expression of breast cancer drug-resistant protein and multidrug-resistant protein 1 in chemoresistant BC cells. Our study identified miR-26b-5p as a tumor suppressor which targets SRGN to sensitize BC cells to chemotherapeutics. These results suggest that miR-26b-5p and SRGN may serve as potential biomarkers and targets for BC chemotherapy.

Stat5 inhibits NLRP3-mediated pyroptosis to enhance chemoresistance of breast cancer cells via promoting miR-182 transcription.

The treatment of breast cancer (BC) calls for targeted methods to overcome chemoresistance (CR). This study is expected to figure out the mechanism of signal transducer and activator of transcription 5 (STAT5) in NOD-like receptor family pyrin domain containing 3 (NLRP3)-mediated pyroptosis and CR in BC cells. BC cell lines resistant to paclitaxel (PTX) and cis-diamminedichloro-platinum (DDP) were prepared. Expressions of Stat5, miR-182, and NLRP3 were detected. The 50% inhibition concentration (IC50 ), proliferation, colony formation, apoptosis rate, and levels of pyroptosis-related factors were appraised and determined. The binding relationships of Stat5 and miR-182, and miR-182 and NLRP3 were testified. Stat5 and miR-182 were highly expressed in drug-resistant BC cells. Silencing Stat5 reduced proliferation and colony formation of drug-resistant BC cells, coincided with elevated levels of pyroptosis-related factors. Stat5 bound to the promoter region of miR-182 to promote miR-182 expression. miR-182 inhibition reversed the role of silencing Stat5 in BC cells. miR-182 inhibited NLRP3. Overall, Stat5 bound to the promoter region of miR-182 to promote miR-182 expression and inhibit NLRP3 transcription, thereby suppressing pyroptosis and enhancing CR of BC cells.

miRNA-144 targeting DNAJC3-AS1 reverses the resistance of the breast cancer cell line Michigan Cancer Foundation-7 to doxorubicin.

This study investigated the role of miRNA-144 (miR-144) targeting of the long noncoding DNAJC3-AS1 in regulating breast cancer chemosensitivity. Real-time quantitative polymerase chain reaction was employed to detect the levels of miR-144 in different drug-resistant cells. MTT assays were used to measure the proliferation of cells in different treatment groups. The apoptosis rate of transfected cells was detected by flow cytometry. Western blotting was used to detect levels of DNAJC3-AS1 protein and of autophagy-related proteins. A double luciferase report experiment was performed to evaluate the targeting effect of miR-144 on DNAJC3-AS1. The level of miR-144 was significantly downregulated in MCF-7 doxorubicin-resistant cells. Upregulated expression of miR-144 increased the doxorubicin sensitivity of drug-resistant cells and the rate of apoptosis. DNAJC3-AS1 was the direct target of miR-144; overexpression of DNAJC3-AS1 significantly rescued the apoptosis induced by miR-144 and reversed the inhibition of autophagy by miR-144. Overexpression of miR-144 can reduce drug resistance in breast cancer cells by inhibiting autophagy or targeting DNAJC3-AS1 for downregulation. miR-144/DNAJC3-AS1 provide a new target for reducing drug resistance in breast cancer.

Dynamics of Plasma EGFR T790M Mutation in Advanced NSCLC: A Multicenter Study.

BACKGROUND: Droplet digital polymerase chain reaction (ddPCR) is an emerging technology for quantitative cell-free DNA oncology applications. However, a ddPCR assay for the epidermal growth factor receptor (EGFR) p.Thr790Met (T790M) mutation suitable for clinical use remains to be established with analytical and clinical validations. OBJECTIVE: We aimed to develop and validate a new ddPCR assay to quantify the T790M mutation in plasma for monitoring and predicting the progression of advanced non-small-cell lung cancer (NSCLC). METHODS: Specificity of the ddPCR assay was evaluated with genomic DNA samples from healthy individuals. The inter- and intraday variations of the assay were evaluated using mixtures of plasmid DNA containing wild-type EGFR and T790M mutation sequences. We assessed the clinical utility of the T790M assay in a multicenter prospective study in patients with advanced NSCLC receiving tyrosine kinase inhibitor (TKI) treatment by analyzing longitudinal plasma DNA samples. RESULTS: We set the criteria for a positive call when the following conditions were satisfied: (1) T790M mutation frequency > 0.098% (3 standard deviations above the background signal); (2) at least two positive droplets in duplicate ddPCR reactions. Among the 62 patients with advanced NSCLC exhibiting resistance to TKI treatment, 15 had one or more serial plasma samples that tested positive for T790M. T790M mutation was detected in the plasma as early as 205 days (median 95 days) before disease progression, determined by imaging analysis. Plasma T790M concentrations also correlated with intervention after disease progression. CONCLUSIONS: We developed a ddPCR assay to quantify the T790M mutation in plasma. Quantification of longitudinal plasma T790M mutation may allow noninvasive assessment of drug resistance and guide follow-up treatment in TKI-treated patients with NSCLC. TRIAL REGISTRATION: Clinical Trials.gov identifier: NCT02804100.

Anti-tumor activity of Shikonin against afatinib resistant non-small cell lung cancer via negative regulation of PI3K/Akt signaling pathway.

Acquired resistance of afatinib is a significant challenge for non-small cell lung cancer (NSCLC) therapy and the mechanisms remain unclear. Aberrant activation of epidermal growth factor receptor (EGFR)-dependent downstream pathways, especially phosphatidylinositol-3-kinases/protein kinase B (PI3K/Akt) signaling pathway has been reported to be involved in the occurrence of afatinib resistance. Developing effective anti-cancer agents to overcome afatinib resistance by targetting PI3K/Akt signaling pathway will be a potential strategy for NSCLC treatment. Shikonin is a naphthoquinone compound isolated from the roots of Lithospermum erythrorhizon In the present study, the anti-cancer activity of Shikonin was evaluated on afatinib-resistant NSCLC in vitro and in vivo The data showed that Shikonin inhibited the proliferation and induced apoptosis of afatinib-resistant NSCLC cell line by activating apoptosis signaling pathway and negatively regulating PI3K/Akt signaling pathway. These results revealed that Shikonin was a potential apoptosis inducer in afatinib-resistant NSCLC and a promising candidate for treating patients clinically.

LncRNA-MALAT1 contributes to the cisplatin-resistance of lung cancer by upregulating MRP1 and MDR1 via STAT3 activation.

Multiple drug resistance is the main reason for chemotherapeutic failure in lung cancer patients with complex molecular mechanisms. LncRNA-MALAT1 plays functional roles in the progression of carcinomas and development of drug resistance. We aimed to identify the role of MALAT1 in DDP-resistant non-small cell lung cancer as well as potential mechanisms. Human lung cancer cell line A549 and the DDP-resistant cell line A549/DDP were used. Cell transfection was performed to establish A549/MALAT1 and A549/DDP/shMALAT1 cells. The qRT-PCR analysis was performed to detect lncRNA-MALAT1 level. Cell viability, colony formation assay, apoptosis analysis, western blot analysis, immunohistochemistry, and animal study were carried out. MALAT1 was upregulated in DDP-resistant A549 cell line. MALAT1 decreased DDP sensitivity in vitro and in vivo by upregulating MRP1 and MDR1 via STAT3 activation. Overexpression of MALAT1 contributed to the DDP resistance and might confer a potently poor prognosis.

Diagnostic Value of Volume-Based Fluorine-18-Fluorodeoxyglucose PET/CT Parameters for Characterizing Thyroid Incidentaloma.

Objective: To assess clinical value of fluorine-18-fluorodeoxyglucose positron emission tomography/computed tomography (PET/CT) for differentiation of malignant from benign focal thyroid incidentaloma. Materials and Methods: This retrospective study included 99 patients with focal thyroid incidentaloma of 5216 non-thyroid cancer patients that had undergone PET/CT. PET/CT semi-quantitative parameters, volume-based functional parameters, metabolic tumor volume (MTV), and total lesion glycolysis (TLG) of thyroid incidentaloma were assessed. Receiver-operating characteristic (ROC) analysis was conducted and areas under the curve (AUC) were compared by Hanley and McNeil test to evaluate usefulness of maximum standardized uptake value (SUVmax), MTV and TLG, as markers for differentiating malignant from benign thyroid incidentalomas. Results: Of 99 thyroid incidentalomas, 64 (64.6%) were malignant and 35 (35.4%) were benign. Malignant thyroid incidentalomas were larger (1.8 cm vs. 1.3 cm, p = 0.006), and had higher SUVmax (11.3 vs. 4.8, p < 0.001), MTV (all p < 0.001) and TLG (all p < 0.001) than benign. TLG 4.0 had the highest performance for differentiation of malignant from benign thyroid incidentaloma in all semi-quantitative parameters with AUC 0.895 by ROC curve analysis. AUC (TLG 4.0) was significantly larger than AUC (SUVmean), AUC (MTV 2.5), AUC (MTV 3.0), AUC (MTV 3.5), AUC (TLG 2.5), and AUC (TLG 3.0), respectively (all, p < 0.05). There was no statistical difference between AUC (TLG 4.0) and AUC (SUVmax) (p > 0.05). A threshold TLG 4.0 of 2.475 had 81.3% sensitivity and 94.3% specificity for identifying malignant thyroid incidentalomas. Conclusion: Volume-based PET/CT parameters could potentially have clinical value in differential diagnosis of thyroid incidentaloma along with SUVmax.

Trends in Treatment for Prostate Cancer in China: Preliminary Patterns of Care Study in a Single Institution.

Objectives: A Patterns of Care Study (PCS) was performed in the largest regional medical center in Zhejiang Province, China. The hospital information system (HIS) was used to evaluate patient characteristics and changes in initial treatment patterns for prostate cancer and to determine recent predominant trends in treatment plans for prostate cancer (PCa) in China. Methods: Men who were newly diagnosed with localized or locally advanced PCa for 2010-2011 and 2016-2017 were identified in the HIS database. Patient characteristics and temporal trends in initial management were assessed, and differences between groups were evaluated for significance using Chi-square and Mann-Whitney U tests. Results: In total, 1792 patients met the study criteria, including 505 and 1287 patients in the 2010-2011 and 2016-2017 samples, respectively. The average age of patients diagnosed in the 2010-2011 PCS survey was 70 years, decreasing to 68 years when the 2016-2017 patients were included (P<0.001). In the 2010-2011 sample, 50.69% of the patients had an initial prostate-specific antigen (PSA) level ≥20 ng/ml. In contrast, the initial PSA level was 4-19.99 ng/ml for 66.67% of the patients in the 2016-2017 sample (P<0.001). Based on National Comprehensive Cancer Network (NCCN) criteria, the percentages of patients in low- and intermediate-risk groups increased from 33.06% to 54.78%; conversely, the percentages in high-risk, very high-risk, and regional (N1) groups decreased to a certain extent (P<0.001). According to European Association of Urology (EAU) criteria, the percentages of patients in low- and intermediate-risk groups increased from 32.07% to 53.69%, yet the percentage in the high-risk group decreased (P<0.001). The use of radical prostatectomy (RP) and radiation therapy (RT) increased from 48.32% to 76.46% and 5.35% to 16.94%, particularly in high-risk and low-risk groups, respectively, whereas the rates of hormone therapy (HT) and active surveillance and observation (AS&O) decreased from 32.28% to 4.27% and from 16.04% to 2.33%, respectively (P<0.001). A similar pattern was observed when patients were stratified by EAU risk group. Conclusions: The results of this real-world study in the largest regional medical center in Zhejiang Province, China, indicate that the predominant characteristics of PCa patients and trends in initial management are changing rapidly. We found the following: (a) a trend toward a decreased age among newly diagnosed patients; (b) a trend toward lower initial PSA levels; (c) a downward trend in risk group classification; (d) a significant increase in the likelihood of receiving RP, particularly in the high-risk group; (e) an increase in the rate of RP, mostly due to use of the Da Vinci robotic system; (f) a significant increase in the likelihood of receiving RT, especially in the low-risk group; and (g) a decrease in HT and AS&O.

Radiological biomarkers for assessing response to locoregional therapies in hepatocellular carcinoma: From morphological to functional imaging (Review).

Many hepatocellular carcinoma (HCC) patients do not qualify for curative surgical intervention and are instead treated with locoregional therapies (LRTs) including ablative and endovascular therapies. Assessment of imaging response is essential in the management of HCC for determining efficacy of therapy and as a surrogate marker for improved survival. The established morphological image biomarkers for tumor burden measurement continue to be applied, as size measurement can easily be used in clinical practice. However, in the setting of liver-directed LRTs for HCC, simple tumor morphological changes can be less informative and usually appear later than biologic changes. Functional imaging (such as perfusion and diffusion imaging, PET-CT/MR and MR spectroscopy) has the potential to be a promising technique for assessment of HCC response to LRTs. Although promising, none of these functional imaging biomarkers have gone through all the required steps of standardization and validation and established accepted criteria for clinical practice.

Ginsenoside Rg3 promotes cytotoxicity of Paclitaxel through inhibiting NF-κB signaling and regulating Bax/Bcl-2 expression on triple-negative breast cancer.

Taxane-based chemotherapy regimen is the most effective therapeutic strategy for triple-negative breast cancer (TNBC) which is an aggressive subtype of breast cancer with high rate of recurrence and distant metastasis. Ginsenoside Rg3 is isolated from Panax ginseng with anti-cancer activity against carcinomas. We aim to evaluate the chemosensitizing effects of Ginsenoside Rg3 on TNBC cells and xenograft and explore the underlying mechanism. Human triple-negative breast cancer lines MDA-MB-231, MDA-MB-453 and BT-549 were used. Cell viability and survival was detected by MTT assay and colony formation assay. Apoptosis was detected by Annexin V/PI assay and TUNEL. Enzyme-linked immunosorbent assay was performed to determine NF-κB activation. The NF-κB p65, Bcl 2, Bax and Caspase-3 protein expression were detected using Western blot analysis. The results showed that Ginsenoside Rg3 promotes cytotoxicity and apoptosis of Paclitaxel on TNBC cell lines and xenograft. Ginsenoside Rg3 combined Paclitaxel inhibited NF-κB activation, decreased NF-κB p65 and Bcl-2 protein expressions, increased Bax and Caspase-3 protein expressions. The ratio of Bax/Bcl-2 was significantly enhanced by the Ginsenoside Rg3 to Paclitaxel. Ginsenoside Rg3 promotes cytotoxicity and apoptosis of Paclitaxel by inhibiting NF-κB signaling and modulating Bax/Bcl-2 expression on TNBC. Ginsenoside Rg3 should be regarded as a good chemosensitizing agent for TNBC treatment.

Ginsenoside Rg3 enhances the anti-proliferative activity of erlotinib in pancreatic cancer cell lines by downregulation of EGFR/PI3K/Akt signaling pathway.

Erlotinib has shown activity in the management of pancreatic cancer. However, the benefit of EGFR blockade is limited due to EGFR independent PI3K/Akt signaling pathway. Studies have reported that Ginsenoside Rg3 strongly inhibited PI3K-Akt signaling pathway of many carcinomas. We aimed to investigate the activity of Ginsenoside Rg3 to sensitize erlotinib in treating pancreatic cancer in vitro and in vivo. Human pancreatic cancer cell lines BxPC-3 and AsPC-1 were used. Cell proliferation and colony formation assay, Annexin V/PI apoptosis analysis, Western blot analysis, immunohistochemistry and in vivo study were carried out. Ginsenoside Rg3 enhanced the anti-proliferative effects of erlotinib in BxPC-3 and AsPC-1 pancreatic cancer cells and xenograft. Ginsenoside Rg3 enhanced erlotinib-induced apoptosis and increased caspase-3,9 and PARP cleavage expression levels. Erlotinib/Ginsenoside Rg3 treatment decreased the levels of p-EGFR, p-PI3K, and p-Akt expression significantly. Ginsenoside Rg3 could enhance the efficacy of erlotinib to inhibit the proliferation of pancreatic cancer cells via induction of apoptosis and downregulation of the EGFR/PI3K/AKT pathway.