Cell Proliferation Status Research Articles

Preoperative prediction of Ki-67 expression in hepatocellular carcinoma by spectral imaging on dual-energy computed tomography (DECT).

The Ki-67 expression level, which represents the proliferation status of cells, serves as a prognostic marker for hepatocellular carcinoma (HCC); however, the immunochemistry method currently used to evaluate Ki-67 is invasive and is not suitable for patients who have lost the chance for surgery, such as those with advanced tumors or those with other serious organic diseases. This study aimed to investigate the value of quantitative dual-energy computed tomography (DECT) parameters in predicting the expression level of Ki-67 in HCC. This study analyzed the data of 123 consecutive patients with HCC who underwent both Ki-67 immunohistochemistry analysis and two-phase contrast-enhanced DECT imaging. The patients were divided into the following two groups based on the positive rate of Ki-67 (Ki-67%): the high-expression group (Ki-67% >20%, n=74); and the low-expression group (Ki-67% ≤20%, n=49). The computed tomography (CT) values in the 130 and 140 keV monochromatic energy images (HU130-140keV), normalized effective atomic number (NeffZ), fat density (Dfat), and water density (Dwater) were measured and calculated at the arterial phase (AP) and portal venous phase (PVP). A Spearman correlation coefficient analysis, a comparison of parameters between groups, a receiver operating characteristic (ROC) curve analysis for evaluating predictive efficacy, and a multivariable logistic regression analysis were conducted. The NeffZ-AP, HU130 keV-PVP, HU140 keV-PVP, Dfat-PVP, and Dwater-PVP were positively correlated with the Ki-67% (all P<0.05), and the DECT parameter values in the high-expression group were significantly higher than those in the low-expression group (all P<0.05). The Dwater-PVP [odds ratio (OR) =1.353, 95% confidence interval (CI): 1.204-1.521, P<0.001] and tumor diameter (OR =1.258, 95% CI: 1.08-1.465, P=0.003) were independent predictive factors for high Ki-67 expression. Dwater-PVP had the highest predictive efficacy with an area under the curve (AUC) of 0.810. The multivariable analysis combining the DECT parameters and morphological characteristics improved the predictive efficacy of the model in predicting high Ki-67 expression (AUC =0.857). DECT provides a non-invasive method to evaluate the proliferation status of HCC cells, and the predictive efficacy of high Ki-67 expression could be improved by combining DECT parameters and morphologic features.

From the genome's perspective: Bearing somatic retrotransposition to leverage the regulatory potential of L1 RNAs.

Transposable elements (TEs) are mobile genomic elements constituting a big fraction of eukaryotic genomes. They ignite an evolutionary arms race with host genomes, which in turn evolve strategies to restrict their activity. Despite being tightly repressed, TEs display precisely regulated expression patterns during specific stages of mammalian development, suggesting potential benefits for the host. Among TEs, the long interspersed nuclear element (LINE-1 or L1) has been found to be active in neurons. This activity prompted extensive research into its possible role in cognition. So far, no specific cause-effect relationship between L1 retrotransposition and brain functions has been conclusively identified. Nevertheless, accumulating evidence suggests that interactions between L1 RNAs and RNA/DNA binding proteins encode specific messages that cells utilize to activate or repress entire transcriptional programs. We summarize recent findings highlighting the activity of L1 RNAs at the non-coding level during early embryonic and brain development. We propose a hypothesis suggesting a mutualistic relationship between L1 mRNAs and the host cell. In this scenario, cells tolerate a certain rate of retrotransposition to leverage the regulatory effects of L1s as non-coding RNAs on potentiating their mitotic potential. In turn, L1s benefit from the cell's proliferative state to increase their chance to mobilize.

Non-genetic differences underlie variability in proliferation among esophageal epithelial clones.

Individual cells grown in culture exhibit remarkable differences in their growth, with some cells capable of forming large clusters, while others are limited or fail to grow at all. While these differences have been observed across cell lines and human samples, the growth dynamics and associated cell states remain poorly understood. In this study, we performed clonal tracing through imaging and cellular barcoding of an in vitro model of esophageal epithelial cells (EPC2-hTERT). We found that about 10% of clones grow exponentially, while the remaining have cells that become non-proliferative leading to a halt in the growth rate. Using mathematical models, we demonstrate two distinct growth behaviors: exponential and logistic. Further, we discovered that the propensity to grow exponentially is largely heritable through four doublings and that the less proliferative clones can become highly proliferative through increasing plating density. Combining barcoding with single-cell RNA-sequencing (scRNA-seq), we identified the cellular states associated with the highly proliferative clones, which include genes in the WNT and PI3K pathways. Finally, we identified an enrichment of cells resembling the highly proliferative cell state in the proliferating healthy human esophageal epithelium.

Integrating IL-12 mRNA nanotechnology with SBRT eliminates T cell exhaustion in preclinical models of pancreatic cancer

Pronounced T cell exhaustion characterizes immunosuppressive tumors, with the tumor microenvironment (TME) employing multiple mechanisms to elicit this suppression. Traditional immunotherapies, such as immune checkpoint blockade, often fail due to their focus primarily on T cells. To overcome this, we utilized a proinflammatory cytokine, IL-12, that re-wires the immunosuppressive TME by inducing T cell effector function while also repolarizing immunosuppressive myeloid cells. Due to toxicities observed with systemic administration of this cytokine, we utilized lipid nanoparticles encapsulating mRNA encoding IL-12 for intratumoral injection. This strategy has been proven safe and tolerable in early clinical trials for solid malignancies. We report an unprecedented loss of exhausted T cells and the emergence of an activated phenotype in murine pancreatic ductal adenocarcinoma (PDAC) treated with stereotactic body radiation therapy (SBRT) and IL-12mRNA. Our mechanistic findings reveal that each treatment modality contributes to the T cell response differently, with SBRT expanding the TCR repertoire and IL-12mRNA promoting robust T cell proliferation and effector status. This distinctive T cell signature mediated marked growth reductions and long-term survival in local and metastatic PDAC models. This is the first study of its kind combining SBRT with IL-12mRNA and provides a promising new approach for treating this aggressive malignancy.

A transcriptomic biomarker predictive of cell proliferation for use in adverse outcome pathway-informed testing and assessment.

High-throughput transcriptomics (HTTr) is increasingly being used to identify molecular targets of chemicals that can be linked to adverse outcomes. Cell proliferation (CP) is an important key event in chemical carcinogenesis. Here, we describe the construction and characterization of a gene expression biomarker that is predictive of the CP status in human and rodent tissues. The biomarker was constructed from 30 genes known to be increased in expression in prostate cancers relative to surrounding tissues and in cycling human MCF-7 cells after estrogen receptor (ER) agonist exposure. Using a large compendium of gene expression profiles to test utility, the biomarker could identify increases in CP in (i) 308 out of 367 tumor vs. normal surrounding tissue comparisons from 6 human organs, (ii) MCF-7 cells after activation of ER, (iii) after partial hepatectomy in mice and rats, and (iv) the livers of mice and rats after exposure to nongenotoxic hepatocarcinogens. The biomarker identified suppression of CP (i) under conditions of p53 activation by DNA damaging agents in human cells, (ii) in human A549 lung cells exposed to therapeutic anticancer kinase inhibitors (dasatinib, nilotnib), and (iii) in the mouse liver when comparing high levels of CP at birth to the low background levels in the adult. The responses using the biomarker were similar to those observed using conventional markers of CP including PCNA, Ki67, and BrdU labeling. The CP biomarker will be a useful tool for interpretation of HTTr data streams to identify CP status after exposure to chemicals in human cells or in rodent tissues.

Autophagic genes and antioxidant status during doxorubicin therapy under temperature-stressed conditions in breast cancer cells

ObjectivesAutophagy as a cellular event swings between apoptosis induction and cellular maintenance during chemotherapy. The interplay between autophagy and reactive oxygen species (ROS) remains obscure in cancer progression and treatment. This study aimed to determine the combination effect of chemotherapy and hyperthermia on cancer cell proliferation, autophagy signaling, and oxidative stress status. MethodsTo estimate the autophagic genes' role and antioxidant capacity involvement during chemotherapy, human breast cancer cell lines (MCF7 and CAL51) were exposed to a high temperature at 42 °C and treated with doxorubicin (DOX). The MTT assays were performed to determine the cell survival levels, and the autophagic gene expression levels (ATG5, LC3A, LC3B, and Beclin1) were analyzed. The intracellular TAS (total antioxidant status) and SOD (superoxide dismutase) activities were determined. ResultsThe combined effects of high temperature with chemotherapy significantly reduced cell viability and cell survival compared to cells treated with the chemotherapeutic agent DOX alone. The cell's autophagic gene activities were significantly increased after exposure to 42 °C temperature and DOX-based chemotherapy compared to the cells treated with DOX alone. It was observed that TAS and SOD activities were increased in the cell lines exposed to high temperatures with DOX compared to the cells treated with DOX alone. Increased autophagic gene expression levels and cell death were observed in response to the high temperature and DOX treatment in breast cancer cells. ConclusionThe study may provide a plausible route and treatment plan for the individual adoption of cancer chemotherapy and be developed as part of the personalized medicine protocol.

Heat Shock Related Protein Expression in Abdominal Testes of Asian Elephant (Elephas maximus).

The abdominal testes of Asian elephants show normal spermatogenesis. Heat shock in cryptorchid testes elevates heat shock factor (HSF) expression, leading to germ cell apoptosis, while increased heat shock proteins (HSPs) levels provide protection. To investigate how heat shock affects elephant spermatogenic cells, focusing on heat shock-related molecules and the cell death mechanism, immunohistochemistry and TUNEL staining were employed to assess the immunoexpression of several heat shock-related molecules and the status of apoptosis in elephant fibroblasts (EF) induced by heat shock stimulus. Additionally, the immunoexpression of heat shock-related molecules and cell proliferation status in the elephant spermatogenic cells. Our finding indicated that heat shock-induced HSF1 immunoexpression in EF leads to apoptosis mediated by T-cell death-associated gene 51 (TDAG51) while also upregulating HSP70 to protect damaged cells. In elephant spermatogenic cells, immunostaining revealed a predominance of proliferating cell nuclear antigen (PCNA)-positive cells with minimal TDAG51- and TUNEL-positive cells, suggesting active proliferation and apoptosis suppression during normal spermatogenesis in the abdominal testis. Interestingly, spermatogonia co-immunoexpressed HSF1 and HSP90, potentially reducing apoptosis through protective mechanisms different from those observed in other mammals. Spermatogenic cells did not show immunolocalisation of HSP70, and hence, it may not contribute to protecting the spermatogonia from heat shock because the transcriptional activity of HSF1 is suppressed by HSP90A binding. This study provides insight into the specific heat shock response and defence mechanisms in elephant spermatogenic cells and may contribute to our understanding of species-specific adaptation to environmental stresses of the testis.

ALTMAN: A Novel Method for Cell Cycle Analysis.

Accurate analysis of S-phase fraction is crucial for the assessment of cell proliferation levels, tumor malignancy and prognostic effects of treatment. Most of the currently developed methods for S-phase cell analysis rely on flow cytometric analysis of DNA content determination. However, the lack of standardized procedures for sample analysis and interpretation of cell cycle fitting graphs poses a significant limitation in clinical practice for utilizing flow cytometry to measure the cell cycle based on DNA content. Herein, we developed an approach for analyzing S-phase cells based on telomerase activity determination. Briefly, this approach distinguishes S-phase cells in cell populations via direct fluorescence tracking of telomerase activity within individual cells. The dynamic analysis of telomerase activity in different cell cycles was made possible by the ALTMAN strategy developed in our previous studies, which has been successfully employed to distinguish S-phase cells in cultured cells. This method offers a novel avenue for the assessment of cell cycle status and the evaluation of the proliferation status of tumor cells and the prognosis effect of tumor patients via analyzing the differences in telomerase activity during different cell cycle processes.

The relationship between contrast-enhanced computed tomography radiomics features and mitosis karyorrhexis index in neuroblastoma

ObjectiveMitosis karyorrhexis index (MKI) can reflect the proliferation status of neuroblastoma cells. This study aimed to investigate the contrast-enhanced computed tomography (CECT) radiomics features associated with the MKI status in neuroblastoma.Materials and methods246 neuroblastoma patients were retrospectively included and divided into three groups: low-MKI, intermediate-MKI, and high-MKI. They were randomly stratified into a training set and a testing set at a ratio of 8:2. Tumor regions of interest were delineated on arterial-phase CECT images, and radiomics features were extracted. After reducing the dimensionality of the radiomics features, a random forest algorithm was employed to establish a three-class classification model to predict MKI status.ResultsThe classification model consisted of 5 radiomics features. The mean area under the curve (AUC) of the classification model was 0.916 (95% confidence interval (CI) 0.913–0.921) in the training set and 0.858 (95% CI 0.841–0.864) in the testing set. Specifically, the classification model achieved AUCs of 0.928 (95% CI 0.927–0.934), 0.915 (95% CI 0.912–0.919), and 0.901 (95% CI 0.900–0.909) for predicting low-MKI, intermediate-MKI, and high-MKI, respectively, in the training set. In the testing set, the classification model achieved AUCs of 0.873 (95% CI 0.859–0.882), 0.860 (95% CI 0.852–0.872), and 0.820 (95% CI 0.813–0.839) for predicting low-MKI, intermediate-MKI, and high-MKI, respectively.ConclusionsCECT radiomics features were found to be correlated with MKI status and are helpful for reflecting the proliferation status of neuroblastoma cells.

Effect of graphene oxide loaded on TiO2-nanotube-modified Ti on inflammatory responses

Although a titanium matrix modified with titanium dioxide nanotube (TNT) arrays can have anti-inflammatory effects in vitro, these effects are limited. In this study, the TNT surface was modified by electrodepositing graphene oxide (GO) to enhance the anti-inflammatory effect of the material. Scanning electron microscopy (SEM), Raman spectroscopy, and x-ray diffraction were used to characterize each of these materials. Cell Counting Kit-8 (CCK-8) was used to determine the cell proliferation status. Enzyme-linked Immunosorbent Assay (ELISA), immunofluorescence staining, and RNA sequencing were used to assess the regulation of inflammation in each group. Raman spectroscopy confirmed that GO was successfully loaded onto the surface. The SEM, ELISA, fluorescence staining, and RNA sequencing results indicated that TNT-GO can effectively inhibit the inflammatory response and induce the M2 polarization of macrophages. TNT-GO can weaken the surface inflammatory responses of materials, suppress the secretion of pro-inflammatory factors, and promote the M2 polarization of macrophages. These advantageous properties render TNT-GO a promising material for dental implants.

SIRT3 protects endometrial receptivity in patients with polycystic ovary syndrome.

The sirtuin family is well recognized for its crucial involvement in various cellular processes. Nevertheless, studies on its role in the human endometrium are limited. This study aimed to explore the expression and localization of the sirtuin family in the human endometrium, focusing on sirtuin 3 (SIRT3) and its potential role in the oxidative imbalance of the endometrium in polycystic ovary syndrome (PCOS). Endometrial specimens were collected from both patients with PCOS and controls undergoing hysteroscopy at the Center for Reproductive Medicine, Peking University Third Hospital, from July to August 2015 and used for cell culture. The protective effects of SIRT3 were investigated, and the mechanism of SIRT3 in improving endometrial receptivity of patients with PCOS was determined using various techniques, including cellular bioenergetic analysis, small interfering ribonucleic acid (siRNA) silencing, real-time quantitative polymerase chain reaction, Western blot, immunofluorescence, immunohistochemistry, and flow cytometry analysis. The sirtuin family was widely expressed in the human endometrium, with SIRT3 showing a significant increase in expression in patients with PCOS compared with controls (P <0.05), as confirmed by protein and gene assays. Concurrently, endometrial antioxidant levels were elevated, while mitochondrial respiratory capacity was reduced, in patients with PCOS (P <0.05). An endometrial oxidative stress (OS) model revealed that the downregulation of SIRT3 impaired the growth and proliferation status of endometrial cells and reduced their receptivity to day 4 mouse embryos. The results suggested that SIRT3 might be crucial in maintaining normal cellular state by regulating antioxidants, cell proliferation, and apoptosis, thereby contributing to enhanced endometrial receptivity. Our findings proposed a significant role of SIRT3 in improving endometrial receptivity in patients with PCOS by alleviating OS and regulating the balance between cell proliferation and apoptosis. Therefore, SIRT3 could be a promising target for predicting and improving endometrial receptivity in this patient population.

Exploring a multiparameter MRI-based radiomics approach to predict tumor proliferation status of serous ovarian carcinoma.

To develop a multiparameter magnetic resonance imaging (MRI)-based radiomics approach that can accurately predict the tumor cell proliferation status of serous ovarian carcinoma (SOC). A total of 134 patients with SOC who met the inclusion and exclusion criteria were retrospectively screened from institution A, spanning from January 2016 to March 2022. Additionally, an external validation set comprising 42 SOC patients from institution B was also included. The region of interest was determined by drawing each ovarian mass boundaries manually slice-by-slice on T2-weighted imaging fat-suppressed fast spin-echo (T2FSE) and T1 with contrast enhancement (T1CE) images using ITK-SNAP software. The handcrafted radiomic features were extracted, and then were selected using variance threshold algorithm, SelectKBest algorithm, and least absolute shrinkage and selection operator. The optimal radiomic scores and the clinical/radiological independent predictors were integrated as a combined model. Compared with the area under the curve (AUC) values of each radiomic signature of T2FSE and T1CE, respectively, the AUC value of the radiomic signature (T1CE-T2FSE) was the highest in the training set (0.999 vs. 0.965 and 0.860). The homogeneous solid component of the ovarian mass was considered the only independent predictor of tumor cell proliferation status among the clinical/radiological variables. The AUC of the radiomic-radiological model was 0.999. The radiomic-radiological model combining radiomic scores and the homogeneous solid component of the ovarian mass can accurately predict tumor cell proliferation status of SOC which has high repeatability and may enable more targeted and effective treatment strategies. The proposed radiomic-radiological model combining radiomic scores and the homogeneous solid component of the ovarian mass can predict tumor cell proliferation status of SOC which has high repeatability and may guide individualized treatment programs. • The radiomic-radiological nomogram may guide individualized treatment programs of SOC. • This radiomic-radiological nomogram showed a favorable prediction ability. • Homogeneous slightly higher signal intensity on T2FSE is vital for Ki-67.

Cell-to-cell adhesion via CD54 (intercellular adhesion molecule-1)-associated cell proliferation in diffuse large B-cell lymphoma cases.

Cluster of Differentiation 54 (CD54), also known as intracellular adhesion molecule-1 (ICAM-1), is a transmembrane glycoprotein belonging to the immunoglobulin superfamily. Although CD54 has been shown to be involved in cell-to-cell adhesion and proliferation of B-cell lymphoma cell lines, the clinical significance of its expression has not yet been elucidated. We analyzed Ki-67 indices, the expression status of CD54 and its receptor (CD11a), and the intercellular distance of tumor cells in 40 diffuse large B-cell lymphoma (DLBCL) cases with vascular invasion to analyze the association of cell adhesion and proliferation status. CD54 and CD11a were simultaneously expressed (double-positive) in extra/intravascular tumor cells in 14 (35%) of the cases. Histologically, lymphoma cells of the double positive cases exhibited significantly higher Ki-67 index in extravascular tumor cells than that in the intravascular ones, while no difference was observed in lymphoma cells of the non-double positive cases. The significantly shorter extravascular intercellular distance compared with the intravascular intercellular distance suggested the association between cell-cell adhesion mediated by CD54 and cell proliferation. We further confirmed that the treatment of the recombinant LFA1 (CD11a/CD18) showed the adhesion of human DLBCL-derived cell line HBL-2 to LFA1 and increased cell viability. These findings suggest that cell-to-cell adhesion via CD54 maintains the cell proliferative activity of a subset of DLBCL. This study provides a valuable foundation upon which further research may be conducted to determine detailed mechanisms of cell-to-cell-associated and adhesion-independent cell proliferation.

Effect of Cryoprotectants on Long-Term Storage of Oral Mucosal Epithelial Cells: Implications for Stem Cell Preservation and Proliferation Status.

In this study, we tested a method for long-term storage of oral mucosal epithelial cells (OMECs) so that the cells could be expanded in vitro after cryopreservation and used for the treatment of bilateral limbal stem cell deficiency. The ability of suspended primary OMECs to proliferate in vitro after cryopreservation was compared to that of OMEC cultures that had undergone the same process. Both were preserved in standard complex medium (COM) with or without cryoprotective agents (CPAs) (gly-cerol at 5 % or 10 % or dimethyl sulphoxide at 10 %). We found that after cryopreservation, primary OMECs could form a confluent cell sheet only in a few samples after 22 ± 2.9 (mean ± SD) days of cultivation with 72.4 % ± 12.9 % overall viability. Instead, all ex vivo OMEC cultures could re-expand after cryopreservation with a comparable viability of 78.6 ± 13.8 %, like primary OMECs, but with significantly faster growth rate (adj. P &lt; 001), forming a confluent cell sheet at 13.7 ± 3.9 days. Gene expression analyses of the ex vivo expansion of OMEC cultures showed that the stemness, proliferation and differentiation-related gene expression was similar before and after cryopreservation, except for KRT13 expres-sion, which significantly decreased after the second passage (adj. P &lt; 0.05). The addition of CPAs had no effect on these outcomes. In conclusion, the optimal strategy for OMEC preservation is to freeze the cells that have been previously cultured, in order to maintain cell viability and the capacity to create a sizable graft even without CPAs.

Correlation analysis of dynamic enhanced energy spectrum CT parameters with Ki-67 high expression in hepatocellular carcinoma

Objective: To investigate the correlation between dynamic enhanced energy spectral CT parameters and Ki-67 high expression in hepatocellular carcinoma (HCC). Methods: A cross-sectional study. This retrospective case-control study analyzed the clinical data of 101 patients with pathologically confirmed HCC in Xiamen Hospital of Traditional Chinese Medicine and Zhongshan Hospital of Xiamen University from December 2017 to March 2023. These 101 patients included 84 males and 17 females, and the age[M(Q1, Q3)] was 59.0(49.0,66.0)years. These patients were divided into two groups according to the immunohistochemical Ki-67 expression levels in tumor tissues: the high expression group (Ki-67%>20%, n=59) and the low expression group (Ki-67%≤20%, n=42).CT values on 70 keV and 140 keV monochromatic energy images (HU70 keV-a, HU140 keV-a, HU70 keV-p, HU140 keV-p) and water density (Dwater-a, Dwater-p) were measured in arterial phase and portal vein phase, and the difference of HU70 keV, HU140 keV, Dwater values between portal vein and arterial phase (ΔHU70 keV, ΔHU140 keV, ΔDwater), as well as ratio of HU70 keV, HU140 keV, Dwatervalues between portal vein and arterial phase (HU70 keVratio, HU140 keVratio, Dwaterratio) were calculated. Spearman correlation analysis was used to analyze the correlation between the CT spectral parameters and Ki-67%. Multivariate logistic regression model was used to determine the factors associated with high expression of Ki-67. The receiver operating characteristics (ROC) curves were used to indicate the efficacy of dynamic enhanced spectral CT in evaluating Ki-67 high expression in HCC. Results: The high Ki-67 expression group revealed higher alpha fetal protein levels, larger tumor diameter and more irregular tumor shape compared with the low Ki-67 expression group,and the differences were statistically significant (all P<0.05). Spearman correlation analysis showed that the HU140 keV-p, Dwater-p, HU70 keV ratio, HU140 keV ratio, Dwater ratio, ΔHU70 keV, ΔHU140 keV, ΔDwater were positively correlated with Ki-67 positivity rate (r:0.31-0.50, all P<0.05). The spectral CT parameters (HU70 keV-p, HU140 keV-p, Dwater-p, HU70 keV ratio, HU140 keV ratio, Dwater ratio, ΔHU70 keV, ΔHU140 keV, ΔDwater) in high Ki-67 expression group were significantly higher than those in low Ki-67 expression group (all P<0.05). Multivariate logistic regression model shows that Dwater-p(OR=1.16, 95%CI: 1.05-1.29, P=0.005), ΔHU140 keV(OR=1.39, 95%CI: 1.20-1.62, P<0.001) and irregular tumor morphology (OR=5.25, 95%CI: 1.61-17.12, P=0.006) were correlative factors for high Ki-67 expression. The HU140 keV ratio and ΔHU140 keV alone evaluated the highest AUC of high Ki-67 high expression in HCC, which were 0.82 (95%CI: 0.74-0.90), the sensitivity were 61.0%, and the specificity were 88.1% and 85.7%. The combined analysis of Dwater-p, ΔHU140 keV and irregular tumor morphology had an increased AUC of 0.88 (95%CI: 0.81-0.95) in assessment high Ki-67 expression, with the sensitivity of 84.7% and the specificity of 78.6%. Conclusions: Dynamic enhanced spectral CT parameters were positively correlated with the Ki-67 expression in HCC. Spectral CT provides a non-invasive method to evaluate the proliferation status of HCC cells, and the efficiency could be improved by multi-parameter analysis combining spectral CT parameters and morphologic features.

OIP5-AS1/CD147/TRPM7 axis promotes gastric cancer metastasis by regulating apoptosis related PI3K-Akt signaling.

To explore the mechanism of OIP5-AS1/CD147/TRPM7 axis to gastric cancer (GC) metastasis. Bioinformatic analysis was performed to pick up the candidate genes associated with regulation GC metastasis. Using GC cell lines, AGS and MKN-45 as research objects, identify the effect of candidate genes on GC metastasis, judge cell proliferation status by MTT assay and cell clone number, and detect cell migration by Transwell and Wound-healing assay. The molecular mechanism of CD147/OIP5/TRPM7 axis regulating GC metastasis was further explored by RNA sequencing. The key signaling pathways were subsequently verified by flow cytometry and WB. Bioinformatic analysis suggested OIP5-AS1/CD147/TRPM7 axis may be involving in GC metastasis. The RNA interference experiment proved that after gene interference, the proliferation ability of GC cells decreased significantly (P<0.05), which was manifested in the reduction of the number of cell clones. In addition, the migration ability of GC cells was also affected, which was based on the results of Wound Healing (P<0.05). CD147, OIP5-AS1 and TRPM7 all have harmful effects on GC cells. The relationship between OIP5-AS1 and CD147/TRPM7 was detected by RNA immunoprecipitation. Moreover, the RNA sequencing data indicated that CD147/OIP5-AS1/TRPM7 may coordinately regulate the PI3K-AKT pathway related to GC cell apoptosis, thereby affecting the proliferation and migration of GC cells. After RNA interference, the level of apoptosis increased both in AGS and MKN-45 cells. Meanwhile, the expression of pro-apoptotic proteins Caspase9 and BAX were up-regulated (P<0.05). In addition, the expression of PI3K and AKT proteins was reduced (P<0.05). The mouse tumorigenesis experiment corroborated the results of the in vitro study. OIP5-AS1/CD147/TRPM7 axis reduces GC cell proliferation by regulating apoptosis associated with PI3K-AKT signaling, further affecting cancer metastasis.

Cell Line-Based Human Bladder Organoids with Bladder-like Self-Organization-A New Standardized Approach in Bladder Cancer Research.

Three-dimensional tumor models have gained significant importance in bladder cancer (BCa) research. Organoids consisting of different cell types better mimic solid tumors in terms of 3D architecture, proliferation, cell-cell interaction and drug responses. We developed four organoids from human BCa cell lines with fibroblasts and smooth muscle cells of the bladder, aiming to find models for BCa research. The organoids were characterized in terms of cytokeratins, vimentin, α-actin and KI67 by immunoreactivity. Further, we studied ligand-dependent activation of the Wnt/β-catenin pathway and investigated the responses to anti-tumor therapies. The organoids mimicked the structure of an inverse bladder wall, with outside urothelial cells and a core of supportive cells. The cytokeratin staining patterns and proliferation rate were in conjunction with the origins of the BCa cells. RT-112 even showed stratification of the epithelium. Treatment with Wnt10B led to increased β-catenin (active) levels in high-grade organoids, but not in low-grade BCa cells. Doxorubicin treatment resulted in clearly reduced viability (10-30% vs. untreated). In contrast, the effectivity of radiotherapy depended on the proliferation status of BCa cells. In conclusion, cell-line-based organoids can form bladder-like structures and reproduce in vivo features such as urothelial differentiation and stratification. Thus, they can be useful tools for functional studies in BCa and anti-cancer drug development.

Correlation between metabolomic profile and proliferation of Macrobrachium rosenbergii primary embryonic cell culture

Despite extensive research spanning several decades, a commercial cell line for prawns remains elusive, and we have yet to determine the ideal conditions required for successful in vitro cell cultures of crustaceans. This study marks the first application of mass spectrometry to analyze lipid content changes in prawn culture media, allowing us to investigate how specific metabolites in media correlate with cell cycle and proliferation status. To investigate these relationships, short-term cell culture experiments were conducted using three commercial media: Dulbecco's modified eagle medium (DMEM) high glucose, Opti-Minimal essential media (Opti-MEM), and DMEM:F12. We monitored metabolic activity, performed cell-cycle analysis, and used mass spectrometry to analyze lipid metabolic profiles of culture media. This comprehensive approach enables us to examine the interplay between media lipidomic content and cellular behavior. Our results revealed that all three media sustained cellular metabolic activity. DMEM high glucose and Opti-MEM showed peaks in metabolic activity at day 4, while DMEM:F12 exhibited a peak at day 7. Opti-MEM supported the shortest doubling time, making it ideal for short-term culture. Interestingly, DMEM high glucose retained more attached cells on day 1 and had the highest count on day 4, with a noticeable trend towards the G2/M cell-cycle phase. Moreover, our lipidomic analysis of Opti-MEM revealed significantly higher concentrations of 26 annotated lipids compared to the other media. Notably, lipids like phosphatidylcholine and phosphatidylserine were consistently abundant in all three media, and palmitate levels correlated with proliferation phase. These findings have significant implications for establishing sustainable crustacean cell culture lines. Understanding the lipidomic compositions and their associations with cell-cycle dynamics and proliferation allows for precise conditioning of culture media to promote sustainable growth and delay cellular entry into quiescence phases. This knowledge contributes to the long-term goal of maintaining robust prawn cell cultures for various research and biotechnological applications.

Eribulin promotes proliferation of CD8+ T cells and potentiates T cell-mediated anti-tumor activity against triple-negative breast cancer cells.

Chemotherapeutic agents exert immunomodulatory effects on triple-negative breast cancer (TNBC) cells and immune cells. Eribulin favorably affects the immunological status of patients with breast cancer. However, the effects of eribulin on the immune cells remain unexplored. The aim of this study was to investigate the effects of eribulin on immune cells. Peripheral blood mononuclear cells (PBMCs) from healthy donors and mouse splenocytes were stimulated with anti-CD3 and anti-CD28 antibodies. The effects of eribulin and paclitaxel on cell proliferation and differentiation status were analyzed using flow cytometry. RNA sequencing was performed to assess alterations in gene expression in CD8+ T cells following eribulin and paclitaxel treatment. Using TNBC cell lines (MDA-MB-231, Hs578T, and MDA-MB-157), the anti-tumor activity of CD3/CD28-stimulated T cells combined with eribulin or paclitaxel was evaluated. Eribulin did not affect CD3/CD28-stimulated PBMCs proliferation. However, eribulin significantly decreased the CD4/CD8 ratio in T cells, indicating that eribulin facilitates CD8+ T cell proliferation. Furthermore, eribulin significantly increased the frequency of less differentiated CD45RA+, CCR7+, and TCF1+ subsets of CD8+ T cells. RNA sequencing revealed that eribulin enhanced the expression of gene sets related to cell proliferation and immune responses. Moreover, eribulin augmented the anti-tumor effects of CD3/CD28-stimulated T cells against TNBC cells. These results were not observed in experiments using paclitaxel. Eribulin promoted CD8+ T cell proliferation, repressed effector T cell differentiation, and harnessed T cell-mediated anti-tumor effects. These mechanisms may be one of the cues that eribulin can improve the immunological status of tumor-bearing hosts.

Glial Cell Adhesion Molecule (GlialCAM) Determines Proliferative versus Invasive Cell States in Glioblastoma.

The malignant brain cancer glioblastoma (GBM) contains groups of highly invasive cells that drive tumor progression as well as recurrence after surgery and chemotherapy. The molecular mechanisms that enable these GBM cells to exit the primary mass and disperse throughout the brain remain largely unknown. Here we report using human tumor specimens and primary spheroids from male and female patients that glial cell adhesion molecule (GlialCAM), which has normal roles in brain astrocytes and is mutated in the developmental brain disorder megalencephalic leukoencephalopathy with subcortical cysts (MLC), is differentially expressed in subpopulations of GBM cells. High levels of GlialCAM promote cell-cell adhesion and a proliferative GBM cell state in the tumor core. In contrast, GBM cells with low levels of GlialCAM display diminished proliferation and enhanced invasion into the surrounding brain parenchyma. RNAi-mediated inhibition of GlialCAM expression leads to activation of proinvasive extracellular matrix adhesion and signaling pathways. Profiling GlialCAM-regulated genes combined with cross-referencing to single-cell transcriptomic datasets validates functional links among GlialCAM, Mlc1, and aquaporin-4 in the invasive cell state. Collectively, these results reveal an important adhesion and signaling axis comprised of GlialCAM and associated proteins including Mlc1 and aquaporin-4 that is critical for control of GBM cell proliferation and invasion status in the brain cancer microenvironment.SIGNIFICANCE STATEMENT Glioblastoma (GBM) contains heterogeneous populations of cells that coordinately drive proliferation and invasion. We have discovered that glial cell adhesion molecule (GlialCAM)/hepatocyte cell adhesion molecule (HepaCAM) is highly expressed in proliferative GBM cells within the tumor core. In contrast, GBM cells with low levels of GlialCAM robustly invade into surrounding brain tissue along blood vessels and white matter. Quantitative RNA sequencing identifies various GlialCAM-regulated genes with functions in cell-cell adhesion and signaling. These data reveal that GlialCAM and associated signaling partners, including Mlc1 and aquaporin-4, are key factors that determine proliferative and invasive cell states in GBM.