STK32C modulates doxorubicin resistance in triple-negative breast cancer cells via glycolysis regulation.

Understanding the mechanisms underlying doxorubicin resistance in triple-negative breast cancer (TNBC) holds paramount clinical significance. In our study, we investigate the potential of STK32C, a little-explored kinase, to impact doxorubicin sensitivity in TNBC cells. Our findings reveal elevated STK32C expression in TNBC specimens, associated with unfavorable prognosis in doxorubicin-treated TNBC patients. Subsequent experiments highlighted that STK32C depletion significantly augmented the sensitivity of doxorubicin-resistant TNBC cells to doxorubicin. Mechanistically, we unveiled that the cytoplasmic subset of STK32C plays a pivotal role in mediating doxorubicin sensitivity, primarily through the regulation of glycolysis. Furthermore, the kinase activity of STK32C proved to be essential for its mediation of doxorubicin sensitivity, emphasizing its role as a kinase. Our study suggests that targeting STK32C may represent a novel therapeutic approach with the potential to improve doxorubicin's efficacy in TNBC treatment.

SPARC Controls Migration and Invasion of Hepatocellular Carcinoma Cells Via Regulating GPD2-Mediated Mitochondrial Respiration.

Mitochondrial respiration and metabolism play a pivotal role in facilitating the migratory and invasive capacities of cancer cells. In this study, we aimed to explore the potential influence of glycoprotein SPARC on mitochondrial respiration and its subsequent influence on the migration and invasion of hepatocellular carcinoma (HCC) cells. Lentivirus-mediated shRNA delivery was employed to deplete SPARC in HCC cell lines. The mitochondria localization of SPARC was validated using cellular fractionation followed by Western blot analysis, as well as immunofluorescence staining and Proteinase K protection assay. Co-immunoprecipitation was employed to investigate the interaction between SPARC and GPD2. Seahorse XF Cell Mito Stress Test was conducted to assess the mitochondrial respiration and functionality of HCC cells. Our study identifies an active pool of SPARC within the mitochondria of HCC cells, with the mitochondrial subset proving crucial for the regulation of migration and invasion. The mitochondrial SPARC interacts with GPD2, influencing its expression levels and subsequently modulating GPD2-mediated mitochondrial respiration. This regulatory mechanism orchestrates the migratory and invasive phenotypes of HCC cells. Notably, SPARC and GPD2 exhibit upregulated expression in HCC tissues compared to normal liver tissues. High expression levels of both SPARC and GPD2 in HCC patients are associated with a poorer prognosis. Our study unveils a novel role for SPARC in governing HCC cell migration and invasion through regulating GPD2-mediated mitochondrial respiration. These findings underscore the importance of mitochondrial processes in cancer progression and propose the SPARC/GPD2 axis as a promising target for HCC interventions.

Optimization of uniformity in coronary artery enhancement using a bolus tracking technique with a dual region of interest in coronary computed tomographic angiography.

BACKGROUND: Consistent coronary artery enhancement is essential to achieve accurate and reproducible quantification of coronary plaque composition. PURPOSE: To optimize coronary artery uniformity of enhancement using a bolus tracking technique with a dual region of interest (ROI) in coronary computed tomography angiography (CCTA) on a 320-detector CT scanner. MATERIAL AND METHODS: This prospective study recruited 100 consecutive patients who underwent CCTA and were randomly divided into two groups, namely, a manual trigger group (n = 50), in which a manual fast start technique was used to start the diagnostic scan with the visual evaluation of attenuation in the left atrium and left ventricle, and an automatic trigger group (n = 50), in which a bolus tracking technique was used to automatically start the breath-holding command and diagnostic scan with two ROIs placed in the right and left ventricles. Coronary artery image quality was assessed using quantitative and qualitative scores. The enhancement uniformity was characterized by attenuation variability of the ascending aorta (AAO) and coronary arteries. RESULTS: No statistically significant differences in the image quality of the coronary arteries were observed between the two groups (all P > 0.05). The coefficients of variation (COVs) of arterial attenuation in the automatic trigger group were significantly smaller than in the manual trigger group (AAO: 9.89% vs. 17.93%; LMA: 10.35% vs. 18.98%; LAD proximal: 12.09% vs. 20.84%; LCX proximal: 11.85% vs. 20.95%; RCA proximal: 12.13% vs. 20.84%; all P < 0.05). CONCLUSION: The automatic trigger technique accompanied with dual ROI provides consistent coronary artery enhancement and optimizes coronary artery enhancement uniformity in CCTA on a 320-detector CT scanner.

Coronary artery calcium scoring assessment in ultra-low-dose chest computed tomography.

OBJECTIVES: To investigate the effect of non-electrocardiogram (ECG) -triggered ultra-low-dose CT (ULD-CT) with different reconstruction protocols on coronary artery calcium (CAC) scoring assessment, compared with ECG-triggered CAC CT (CAC-CT). METHODS: This prospective study included 115 patients who underwent CAC-CT and ULD-CT scans under the same topogram images. CAC-CT adopted a prospective ECG-triggered sequential acquisition with a tube potential of 120 kV, and the reconstruction protocol was standard Qr36 + slice 3 mm (CACQr-3mm group). ULD-CT adopted a non-ECG-triggered high-pitch acquisition with a tube potential of Sn100 kV, and four groups of images (named ULDQr-3mm, ULDSa-3mm, ULDQr-1.5mm, and ULDSa-1.5mm) were reconstructed using different reconstruction algorithms (standard Qr36, kV-independent Sa36) and slice thicknesses (3 mm, 1.5 mm). The accuracy of CAC detection by ULD-CT was calculated. The agreement of the CAC score between ULD-CT and CAC-CT scans was assessed using intraclass correlation coefficients (ICC) and Bland-Altman plot, and the agreement of risk categorization was assessed using weighted kappa. RESULTS: The sensitivity and specificity of the ULDSa-1.5mm group for detecting positive CAC were 100% and 97.4%, respectively (k = 0.980). The CAC score for the ULDSa-3mm and ULDSa-1.5mm groups demonstrated excellent agreement with the CACQr-3mm group (ICC = 0.992, 0.990, respectively), with a mean difference of -12.3 and - 12.4. The agreement of risk categorization based on absolute and percentile CAC score between the ULDSa-1.5mm and CACQr-3mm groups was excellent (weighted k = 0.954, 0.983, respectively), and risk reclassification rates were low (3.5%, 2.8%, respectively). The effective dose was reduced by approximately 77.2% for the ULD-CT compared to the CAC-CT (0.18 mSv vs. 0.79 mSv, p < 0.001). CONCLUSION: Reconstruction with a 1.5-mm slice thickness and kV-independent iterative algorithmic protocol in ULD-CT yielded excellent agreement in CAC score quantification and risk categorization compared with ECG-triggered CAC-CT.

SMAGP regulates doxorubicin sensitivity in triple-negative breast cancer cells via modulating mitochondrial respiration.

Doxorubicin-based chemotherapy remains as a major therapeutic approach for patients with triple-negative breast cancer (TNBC). However, insensitivity or resistance to doxorubicin treatment limits the therapeutic efficacy. Mitochondrial respiration plays a critical role in regulating the sensitivity of cancer cells to chemotherapy drugs. Here, we found that small trans-membrane and glycosylated protein (SMAGP) is upregulated in TNBC cells in comparison to normal breast and other subtypes of breast cancer cells. High SMAGP expression is associated with poorer overall survival of TNBC patients. Importantly, loss of SMAGP enhanced the sensitivity of TNBC cells to doxorubicin treatment. Mechanistically, we detected a functional pool of SMAGP in the mitochondria of TNBC cells controlling doxorubicin sensitivity via regulating mitochondrial respiration. Thus, our data suggest that SMAGP acts as a novel regulator of doxorubicin sensitivity in TNBC, identifying SMAGP as a promising therapeutic target for improving the efficacy of doxorubicin-based chemotherapy in TNBC patients.

Size-Specific Dose Estimates Based on Water-Equivalent Diameter and Effective Diameter in Computed Tomography Coronary Angiography.

BACKGROUND To determine the difference in size-specific dose estimates (SSDEs), separately based on effective diameter (deff) and water equivalent diameter (dw) of the central slice of the scan range in computed tomography coronary angiography (CTCA). MATERIAL AND METHODS There were 134 patients who underwent CTCA examination, were electronically retrieved. SSDEs (SSDEdeff and SSDEdw) were calculated using 2 approaches: deff and dw. The median SSDEs and mean absolute relative difference of SSDEs were calculated. Linear regression model was used to assess the absolute relative difference of SSDEs based on the ratio of deff to dw. RESULTS The median values of SSDEdeff and SSDEdw were 18.26 mGy and 20.56 mGy, respectively (P<0.01). The former was about 10.08% smaller than the latter. The mean absolute relative difference of SSDEs was 10.48%, ranging from 0.33% to 24.16%. A considerably positive correlation was found between the absolute relative difference of SSDEs and the ratio of deff to dw (R²=0.9561, r=0.979, P<0.01). CONCLUSIONS The value of SSDEdeff was smaller by an average of about 10.08% than SSDEdw in CTCA, and the absolute relative difference increased linearly with the ratio of effective diameter to water equivalent diameter.

Comparative Study of Volume Computed Tomography Dose Index and Size-Specific Dose Estimate Head in Computed Tomography Examination for Adult Patients Based on the Mode of Automatic Tube Current Modulation.

BACKGROUND The aim of this study was to compare the metrics of volume computed tomography index (CTDIvol) and size-specific dose estimate (SSDE), and quantity the differences in head CT examinations of adult patients. MATERIAL AND METHODS A total of 157 patients underwent head CT examination were enrolled in this retrospective study. Pearson correlation analysis and linear regression correlation analysis were performed to observe the correlation between the dose metrics of CTDIvol and SSDEaver versus tube current product (mAs) and water equivalent diameter (WED). Correlated factors of CTDIvol and SSDEaver were analyzed by multivariate linear stepwise regression analysis. RESULTS A sum of 4239 data settings were measured: slices with WED >16 cm was 71.05%, and the slices with f <1 was 72.64%. The average value of the absolute difference between WED and the diameter of AAPM head phantom was 2.24±1.42 cm. Statistically significant difference was found between the values of CTDIvol and SSDEaver (P=0.000). The dispersion degree of the CTDIvol values was greater than that of SSDEaver. Strong positive correlation was shown between CTDIvol and mAs (P=0.000), as well as CTDIvol and WED (P=0.000). Strong positive correlation was shown between SSDEaver and mAs (P=0.000), and moderate correlation for SSDEaver and WED (P=0.000). Both the metrics of mAs and WED were included in the multivariate linear stepwise regression equation to observe the effect of related factors on the value of SSDEaver. CONCLUSIONS SSDEaver with better representative can reproduce the radiation dosage of the specific adult patients in head CT examination.

[Effect of the C-terminal truncated human apoptosis-inducing factor δ1-480 on biological behaviors of MCF-7 cells].

OBJECTIVE: To observe the expression of the C-terminal truncated human apoptosis-inducing factor (AIF) and its biological effect on MCF-7 cells. METHODS: PcDNA3.0-FDT-AIFδ1-480 was transfected into human breast carcinoma MCF-7 cells with lipofectamine. The expression of the truncated AIF gene was detected by Western blotting, and its effects on the biological behaviors of MCF-7 cells and on the expression of cytochrome c (cytC) were evaluated using flow cytometry, MTT assay, colony-forming assay, and mitochondrial membrane potential measurement. RESULTS: PcDNA3.0-FDT-AIFδ1-480 enhanced AIF expression in MCF-7 cells, obviously inhibited the cell proliferation, and significantly reduced the mitochondrial membrane potentials (P<0.05). Transfection of the cells with PcDNA3.0-FDT-AIFδ1-480 promoted the expression of cytC and resulted in significantly increased apoptosis of MCF-7 cells (P<0.05). CONCLUSION: The expression of C-terminal truncated human AIF gene can induce apoptosis of human MCF-7 cells by promoting cytC release from mitochondria.

Enhanced immunotherapeutic effect of modified HPV16 E7-pulsed dendritic cell vaccine by an adeno-shRNA-SOCS1 virus.

Cervical cancer is the second most common cause of cancer-related deaths among women worldwide. However, no efficient therapy exists against cervical cancer and current treatments have several disadvantages. One possible novel approach is to develop immune-based strategies using tumor antigen-loaded dendritic cells (DCs) for the induction of cellular antitumor immunity. In this study, we created a modified HPV16 E7, HPV16mE7, to reduce its transformation activity and to enhance its antigenicity. The siRNA delivery technique was used to silence the suppressor of cytokine signaling 1 (SOCS1) gene in DCs. BM-derived DCs infected by ad-shRNA-SOCS1 were pulsed with the HPV16mE7 protein and then were transfused into mouse models bearing TC-1 tumor cells expressing HPV16 E6/E7. IFN-γ, cytokine (TNF-α, IL-12, IL-6) expression, anti-E7 antibody and cytotoxic T lymphocyte (CTL) levels were measured. The survival rate, survival days and the tumor volume of the mouse models from the different treatment groups were monitored. The data showed that the mE7-pulsed DC vaccine enhanced by adenovirus-mediated SOCS1 silencing exhibited better immunotherapeutic effect on the allografted tumor mouse models. The method by silencing SOCS1 in HPV16mE7 protein-pulsed DCs may provide a new strategy for the development of safe and effective immunotherapy for cervical cancer.

[Establishment of primary Her-2 over- expression human breast cancer cell model].

AIM: Through isolation and purification pri-mary HER2 overexpression human breast cancer cells from malignant pleural effusion and identification the HER2 expression level of the cells to establish the primary HER2 overexpression human breast cancer cell model. METH-ODS: Malignant pleural effusion of HER2 overexpression breast cancer patient was collected. The primary cells were extracted from malignant pleural effusion by Lymphocyte separation medium and the method of density gradient centrifugation. When the primary cells were cultured and spreaded to the 5th generation, the HER2 expression level of the primary cells were detected by the methods of Q-PCR,Western blot and flow cytometry (FCM). Ability of tumor-bearing was detected by tumor-bearing nude mice assay. RESULTS: The primary HER2 overexpression human breast cancer cells were extracted and identified by the methods of Q-PCR, Western blot and tumor-bearing nude mice assay,even though the FCM showed Negative results. CONCLUSION: The primary HER2 overexpression human breast cancer cell model was established; Identification of primary cells need to be confirmed by different methods.