The spatiotemporal dynamics of spatially variable genes in developing mouse brain revealed by a novel computational scheme.

To understand how brain regions form and work, it is important to explore the spatially variable genes (SVGs) enriched in specific brain regions during development. Spatial transcriptomics techniques provide opportunity to select SVGs in the high-throughput way. However, previous methods neglected the ranking order and combinatorial effect of SVGs, making them difficult to automatically select the high-priority SVGs from spatial transcriptomics data. Here, we proposed a novel computational pipeline, called SVGbit, to rank the individual and combinatorial SVGs for marker selection in various brain regions, which was tested in different kinds of public datasets for both human and mouse brains. We then generated the spatial transcriptomics and immunohistochemistry data from mouse brain at critical embryonic and neonatal stages. The results show that our ranking and clustering scheme captures the key SVGs which coincide with known anatomic regions in the developing mouse brain. More importantly, SVGbit can facilitate the identification of multiple gene combination sets in different brain regions. We identified three dynamical sub-regions which can be segregated by the staining of Sox2 and Calb2 in thalamus, and we also found that Nr4a2 expression gradually segregates the neocortex and hippocampus during the development. In summary, our work not only reveals the spatiotemporal dynamics of individual and combinatorial SVGs in developing mouse brain, but also provides a novel computational pipeline to facilitate the selection of marker genes from spatial transcriptomics data.

Extracellular vesicle-derived LINC00482 induces microglial M2 polarization to facilitate brain metastasis of NSCLC.

Considering the crucial role of long non-coding RNAs (lncRNAs) in non-small cell lung cancer (NSCLC), we tried to analyze the role of extracellular vesicle (EV)-derived LINC00482 in the occurrence of brain metastasis in NSCLC. LINC00482 expression was quantified in EVs isolated from serum samples of NSCLC patients (serum-EVs). Ectopic expression and depletion assays were conducted in the microglial cell line HMC3 co-cultured with serum-EVs and in xenograft mouse models of NSCLC to explore the roles of EV-carried LINC00482. LINC00482 was enriched in serum-EVs and induced M2 polarization of microglial cells HMC3 in vitro. LINC00482 competitively bound to miR-142-3p and upregulated the expression of miR-142-3p target gene TGF-ß1 in HMC3 cells, thus promoting microglial M2 polarization. EV-derived LINC00482-induced M2 microglia promoted the malignant properties of NSCLC cells. In vivo data demonstrated that EVs transmitted LINC00482 to regulate the miR-142-3p/TGF-ß1 axis, induce microglial M2 polarization and affect the pre-metastatic niche, thus enhancing brain metastasis of NSCLC. Overall, suppression of the expression of tumor-derived LINC00482 or LINC00482-containing EVs, may serve as an effective target for contributing to the reduction of brain metastasis of NSCLC.

A simple-to-use nomogram for predicting the risk of radiation pneumonitis in patients with thoracic segment esophageal squamous cell carcinoma.

Background: Radiation pneumonitis (RP) is one of the most severe complications of radiotherapy (RT) or concurrent chemoradiotherapy for thoracic segment esophageal squamous cell carcinoma (TSESCC) with delayed diagnose by conventional computed tomography (CT). The study aimed to develop a nomogram to predict the risk of RP early. Methods: Data was collected from 174 patients with clinicopathologically confirmed TSESCC from October 2013 to June 2020. Procalcitonin (PCT), C-reactive protein (CRP), and interleukin-6 (IL-6) levels in serum were dynamically monitored during radiotherapy. Lasso analysis was used for feature screening before multivariate logistic regression analysis to reduce the multicollinearity of variables. A nomogram combined with biological factors and clinical signs for individualized risk assessment and precise prediction of RP was developed and assed the performance with respect to its calibration, discrimination. Results: Of the 174 patients, 30 patients developed RP (grade ≥2) while 144 patients did not. After variable screening by Lasso analysis and logistics multivariate regression analysis, the predictor variables that were finally retained in the nomogram prediction model included IL-6, CRP, and radiotherapy techniques. The model displayed good discrimination with an area under the curve (AUC) of 0.898 (95% CI: 0.849-0.947), with the sensitivity and specificity of 0.967 and of 0.736, respectively. This model also shows good calibration and clinical practical value. In addition, the study provided a web-based version of the dynamic nomogram to facilitate clinical application. Conclusions: The study provides a nomogram model containing IL-6, CRP, RT techniques, which could be conveniently used for individualized prediction of RP in patients with TSESCC during radiotherapy or concurrent chemoradiotherapy.

Pan-Cancer Analysis on the Oncogenic Role of Programmed Cell Death 10.

Purpose: Programmed cell death factor 10 (PDCD10) is associated with intercellular junction, cytoskeleton organization, cell proliferation, apoptosis, exocytosis, and angiogenesis. However, the role of PDCD10 in human cancer is unclear. This study aims to explore the role of PDCD10 in various tumors and its possible mechanism through bioinformatics analysis. Methods: We verified the expression of the PDCD10 gene based on data from the ONCOMINE, TIMER2.0, and TISDB databases. The correlation of PDCD10 with prognosis of patients with different tumors was analyzed using data from the GEPIA2 database. Proteins bound to PDCD10 were analyzed from the STRING database. PDCD10, PDCD10-binding proteins, and associated candidate genes were analyzed in DAVID for functional and pathway analyses. We also evaluated the immunological, clinical, and genetic aspects of distinct cancers by using TIMER2.0 and the connection between PDCD10 expression and tumor immune subtypes by using TISDB. Single-cell sequencing data from the CancerSEA database were used to characterize cancer cell functional states and generate heat maps. Results: PDCD10 overexpression is linked to certain molecular subtypes of human cancer. Low PDCD10 expression in patients with bladder urothelial carcinoma (BLCA), lung adenocarcinoma (LUAD), liver hepatocellular carcinoma (LIHC), adrenocortical carcinoma (ACC), head and neck squamous cell carcinoma (HNSC), kidney chromophobe carcinoma (KICH), brain lower grade glioma (LGG), pancreatic adenocarcinoma (PAAD), uterine corpus endometrial carcinoma (UCEC), oral squamous cell carcinoma (OSCC), and esophageal adenocarcinoma (ESAD) was correlated with favorable OS, whereas high PDCD10 expression in patients with LUSC, KIRC, READ, SKCM, and THYM was correlated with good prognosis. STRING network prediction results showed that 20 proteins, namely, paxillin (PXN), CCM2 scaffold protein (CCM2), TRAF3 interacting protein 3 (TRAF3IP3), FGFR1 oncogene partner 2 (FGFR1OP2), chromosome 4 open reading frame 19 (C4orf19), suppressor of IKBKE 1 (SIKE1), serine/threonine kinase 25 (STK25), striatin (STRN), protein phosphatase 2 catalytic subunit alpha (PPP2CA), mammalian sterile-20-like kinase 4 (MST4), MOB family member 4 (MOB4), protein phosphatase 2 scaffold subunit Abeta (PPP2R1B), sarcolemma-associated protein (SLMAP), serine/threonine kinase 24 (STK24), striatin 4 (STRN4), STRN3, protein phosphatase 2 scaffold subunit A alpha (PPP2R1A), striatin interacting protein 1 (STRIP1), CTTNBP2 N-terminal like (CTTNBP2NL), and cortactin binding protein 2 (CTTNBP2), can bind to PDCD10. Gene enrichment analysis suggested that PDCD10 is involved in the occurrence of different tumors through the Hippo signalling pathway, RNA transport, mRNA monitoring pathway, endocytosis, and T cell receptor signalling pathway. An inverse relationship was found between PDCD10 expression and cancer-associated fibroblasts in LUSC and TGCT, and PDCD10 expression was strongly connected with immunological subtypes, such as C1 (wound healing), C2 (interferon-gamma dominant), C3 (inflammation), C4 (lymphocyte depletion), C5 (immune silenced), and C6 (TGF-beta dominant). Finally, analysis of single-cell sequencing data revealed that PDCD10 expression is linked to epigenetic reprogramming, DNA repair, cell cycle progression, cell differentiation, inflammation, cell proliferation, cell differentiation, cell invasion, and angiogenesis. Conclusion: The results of our investigation demonstrate that PDCD10 has an oncogenic function in many cancer types. This study provides a reference for future research on antitumor therapeutic targets.

Serum Extracellular Nano-Vesicles miR-153-3p to Identify Micronodular Lung Cancer from Sub-Centimeter Lung Nodules.

Identification of malignancy from sub-centimeter lung nodules (LNs, <1 cm) is core for prevention and treatment of primary phases of lung cancer (LC). The study's purpose was to predict micro-nodular lung cancer (mnLC), such as adenocarcinoma in situ (AIS), micro-invasive adenocarcinoma (MIA), and invasive adenocarcinoma stage 1 (IA1) based on serum extracellular nano-vesicles (sEVs) miR-153-3p. sEVs-miR-153-3p was selected in this study and then used to investigate the expression and efficacy by RT-qPCR in the validation phase, followed by sEVs derived from patients of mnLC, benign lung nodule (BLN), and healthy people groups comprised of 135 recruiters. Further, the study established the prediction model which combined sEVs-miR-153-3p expression with multiple clinical-radiomics features by Logistic algorithms and was authenticated by the area under curve (AUC) phenomenon. Then, a simple-to-use nomogram was developed by Cox proportional-hazards regression modeling for predicting mnLC. Besides, in vitro analyses were performed to demonstrate miR-153-3p/ROCK1 axis in regulating biological mechanisms using LC cell lines. Results demonstrated that sEVs-miR-153-5p's expression values were higher in mnLC patients compared to BLN and healthy people. The prediction model for mnLC was successfully established, utilizing sEVs-miR-153-5p biomarker with significant clinical-radiomics features and yielding an AUC = 0.943 (95%CI: 0.898~0.989, P <0.0001). Moreover, the miR-153-3p and its targeted gene ROCK1 were confirmed as down-regulated in NSCLC cell lines and up-regulated expression respectively. Moreover, the miR-153-3p/ROCK1 axis took part in promoting the epithelial-mesenchymal transition (EMT) signaling pathway and regulated certain biological functions, such as proliferation, migration and invasion of LC cells. Therefore, the sEVs-miR-153-3p associated with radiomics-clinical features showed enormous potential to non-invasively identify malignancy from subcentimeter LNs. miR-153-3p could promote the genesis and progress of LC via EMT signaling pathway, which may serve as a therapeutic target.

Distribution, source identification, and health risk assessment of heavy metals in the soil-rice system of a farmland protection area in Hubei Province, Central China.

Heavy metal contamination in farmland soil is of great concern due to the threat to food security arising from the bioaccumulation of heavy metals in crops planted in contaminated soil, such as rice, corn, and vegetables. Cd is the main contaminant in both paddy soils and rice. The purpose of this study was to reveal the spatial distribution of 8 heavy metals (Cd, Cr, Ni, Cu, Pb, Zn, As, and Hg) in the farmland protection areas in northwestern Hubei Province and to evaluate their pollution status, sources, and health risks. The total amounts of these 8 heavy metal elements in the samples were measured, and the health risk posed by their accumulation in rice was evaluated using the health risk evaluation model recommended by the US Environmental Protection Agency (US EPA). The mean contents of Cd, Ni, Cu, Zn, Hg, and As in soil exceeded the background levels (0.17, 37.3, 30.7, 83.6, 0.077, and 12.3 mg kg-1, respectively) of Hubei Province, and Cd showed the highest enrichment coefficient. The concentration of Cd in 89.1% of samples exceeded the limit values stipulated in the Soil Environmental Quality Risk Control Standard for Soil Contamination of Agricultural Land (Trial) (GB15618-2018). The contents of heavy metals showed dissimilar geographical distribution patterns. The principal component analysis (PCA) results indicated that Cd, Zn, Ni, As, and Cu mainly originated from the application of pesticides and fertilizers; Cr mainly originated from soil texture and pedogenesis; exhaust gas generated during transportation was the point pollution source of Pb; livestock wastewater, manure irrigation, and atmospheric deposition were the main pollution source of Hg. The contents of Ni and Cd in 52.2% and 58.7% of the rice samples, respectively, exceeded the limit values stipulated in the Food Safety National Standards for Contaminants in Foods (GB2762-2017), and the average effective Cd content accounted for 81.9% of the total Cd. The average bioconcentration factor of each heavy metal in rice followed the order Cd >Zn >Hg >As >Ni >Cr >Pb. Cd and As were the main noncarcinogenic contributing factors, accounting for 80.8% of the total noncarcinogenic risk. The carcinogenic risk indexes of Cd, As, and Cr exceeded the risk index threshold of 10-4, indicating a carcinogenic risk to the human body. The highest risks to local residents from heavy metals were found in rice. Cd and As were the main noncarcinogenic and carcinogenic factors and should receive greater attention in risk decision management.

Withdrawal Notice: The Correlation Between ERRα Expression Level and Pathological Grade, Overall Survival, and Cancer Types: A Study on a Cohort of 150 Patients with Breast Cancer

The article has been withdrawn by agreement between the editors and publisher of Recent Patents on Anti-Cancer Drug Discovery. The authors are not responding to the editor's requests to provide the language-edited version.Bentham Science apologizes to the readers of the journal for any inconvenience this may have caused.The Bentham Editorial Policy on Article Withdrawal can be found at https://benthamscience.com/editorial-policies-main.php. Bentham Science Disclaimer: It is a condition of publication that manuscripts submitted to this journal have not been published and will not be simultaneously submitted or published elsewhere. Furthermore, any data, illustration, structure or table that has been published elsewhere must be reported, and copyright permission for reproduction must be obtained. Plagiarism is strictly forbidden, and by submit-ting the article for publication the authors agree that the publishers have the legal right to take appropriate action against the authors, if plagiarism or fabricated information is discovered. By submitting a manuscript, the authors agree that the copyright of their article is transferred to the publishers if and when the article is accepted for publication.

Cross-Scale Integration of Nano-Sized Extracellular Vesicle-Based Biomarker and Radiomics Features for Predicting Suspected Sub-Solid Pulmonary Nodules.

Sub-solid nodules (SSN) are common radiographic findings. Due to possibility of malignancy, further evaluation is urgentlyneeded for prevention and management of lung cancer (LC). This current study enrolled patients with SSN, including LC, benign nodules (BN), and healthy individuals as a control, to discover small extracellular vesicles (sEVs) differentially expressed miRNAs (DEMs) as biomarker by next-generation sequencing (NGS) and validation by RT-qPCR. Through cross-scale integration of validated small-molecule and macro-imaging, the prediction model was developed by logistic algorithms and further interpreted into an easy-to-use Nomogram by Cox-proportional hazards modeling. Present study has discovered various sEVs DEMs and sEVs-miR-424-5p that were selected and validated as novel potential biomarkers for cancerous nodule, namely LC. Furthermore, the 10 radiomics signs and 4 clinical features of SSN were merged with sEVs-miR-424-5p and proceeded in multivariate logistic regression analysis to develop the cross-scale integrated modeling, which yielded a significantly higher area under the curve (AUC). Finally, visualization of an easy-to-use nomogram was invented to potentially predict suspected SSN. sEVs-miR-424-5p could be a novel biomarker for distinguishing SSN from LC and BN populations. Its association with cross-scale fusion of radiomics-clinical features will provide great potential to be an errorless prediction of malignant SSN.

Clinical Management of Risk of Radiation Pneumonia with Serum Markers During the Radiotherapy for Patients with Thoracic Malignant Tumors.

PURPOSE: Risk of radiation pneumonia (RP) could not be effectively detected due to non-specific clinical symptoms in the early stage. The purpose of this investigation was to evaluate serum biomarkers of cytokines interleukin-6 (IL-6), C-reactive protein (CRP) and procalcitonin (PCT) for its early detection in patients with thoracic malignant tumors receiving radiotherapy. PATIENTS AND METHODS: The clinical data of 105 patients with thoracic malignant tumors (lung cancer, esophageal carcinoma and mediastinal tumors) treated by radiotherapy were retrospectively analyzed. The patients were divided into RP group and non-RP group according to the Common Terminology Criteria for Adverse Events (CTCAE 5.0). The serum level of IL-6 was detected by chemiluminescence, and the level of CRP was measured by nephelometry during radiotherapy. The level of PCT, one of the specific indicators to distinguish infection and non-infectious etiologies, was also detected by chemiluminescence. RESULTS: Among 105 patients treated by radiotherapy, 28 developed RP, and the other 77 had no RP. There was no significant difference in the risk of RP between patients' factors (age, sex, PS score, smoking, tumor type) and treatment factors (chemotherapy, V5, GTV dose). However, chronic obstructive pulmonary disease (COPD), V20 and mean lung dose (MLD) were significantly different between the two groups (χ2 = 4.131, 3.986, 7.830, P < 0.05). Furthermore, PCT levels were also found to have insignificant differences between RP group and non-RP group (P > 0.05). However, there were significant differences between the groups in the levels of IL-6 and CRP (P < 0.05). The IL-6 levels significantly increased earlier than that of conventional CT imaging when patients suffering from RP and peaked at 6 weeks during radiotherapy. CRP had a similar change as IL-6. Single cytokine and combination of IL-6 and CRP possessed a good ability to predict RP with the AUC of IL-6 of 0.89±0.04 (95% CI, 0.80-0.95, P<0.001), CRP of 0.87±0.05 (95% CI, 0.78-0.94, P<0.001), IL-6 + CRP of 0.92 ± 0.03 (95% CI, 0.83-0.97, P < 0.001), respectively. CONCLUSION: The combined detection of serum IL-6, CRP and PCT may be an effectual method for early detection and clinical practice management of risk of RP.

Improved survival with higher radiation dose for esophageal squamous cell carcinoma patients treated with definitive chemoradiotherapy.

PURPOSE: The optimal radiation dose for patients with esophageal squamous cell carcinoma (ESCC) has long been debated. We undertook the retrospective study to evaluate the survival impact of high dose vs standard dose in patients with stage II-III esophageal cancer treated with definitive chemoradiotherapy (CRT). RESULTS: A total of 137 patients were included in our study, 63 patients classified as standard-dose group and 74 as high-dose group. For the 63 patients in the standard-dose group, the median PFS and the 1-, 2-, and 3-year PFS rates were 12.6 months, 58.0%, 26.0% and 12.0%, respectively; for the 74 patients in the high-dose group, they were 20.0 months, 80.1%, 31.0% and 20.0%, respectively (P = 0.013). The median OS of the patients in the standard-dose group and high-dose group groups were 19.0 months and 26.6 months, respectively, and the 1-, 2- and 3-year survival rates were 78.0%, 39.0%, and 24.0% , and 89.0%, 61.0%, and 30.0%, respectively (P = 0.037). Besides the rate of grade ≥ 3 acute irradiation esophagitis in the high-dose group (10.5% versus. 2.2%, P < 0.01), there were no significantly differ of treatment-related toxicities between the two groups. MATERIALS AND METHODS: According to the radiation dose, patients from 2010 to 2014 were allocated into either the standard-dose group (50-50.4 Gy) or the high-dose group (≥ 59.4 Gy). Overall survival (OS), progression-free survival (PFS) and treatment-related toxicities were assessed and compared between the two groups. CONCLUSIONS: Our findings suggest that higher radiation dose could perform better outcomes for esophageal squamous cell carcinoma patients.

AtOPR3 specifically inhibits primary root growth in Arabidopsis under phosphate deficiency.

The primary root plays essential roles in root development, nutrient absorption, and root architectural establishment. Primary root growth is generally suppressed by phosphate (P) deficiency in A. thaliana; however, the underlying molecular mechanisms are largely elusive to date. We found that AtOPR3 specifically inhibited primary root growth under P deficiency via suppressing root tip growth at the transcriptional level, revealing an important novel function of AtOPR3 in regulating primary root response to the nutrient stress. Importantly, AtOPR3 functioned to down-regulate primary root growth under P limitation mostly by its own, rather than depending on the Jasmonic acid signaling pathway. Further, AtOPR3 interacted with ethylene and gibberellin signaling pathways to regulate primary root growth upon P deficiency. In addition, the AtOPR3's function in inhibiting primary root growth upon P limitation was also partially dependent on auxin polar transport. Together, our studies provide new insights into how AtOPR3, together with hormone signaling interactions, modulates primary root growth in coping with the environmental stress in Arabidopsis.

The synergistic effect and mechanism of doxorubicin-ZnO nanocomplexes as a multimodal agent integrating diverse anticancer therapeutics.

BACKGROUND: Nanomaterials have emerged as ideal multimodal nanomedicine platforms, each one combining different designs and therapeutic approaches in a single system against cancer. The aim of the current study was to explore the synergistic effect and mechanism of a doxorubicin (Dox)-ZnO nanocomplex as a multimodal drug delivery system, integrating Dox chemotherapy and ZnO-mediated photodynamic therapy, in anticancer therapeutics. METHODS: Dox was loaded onto ZnO nanomaterials, forming complexes with the transition metal Zn to yield the Dox-ZnO nanocomplexes. After culture with SMMC-7721 hepatocarcinoma cells, the cellular uptake was quantitatively detected by flow cytometry and visualized by fluorescence microscopy. The synergistic effects of the different anticancer therapeutic modalities on the proliferation of SMMC-7721 hepatocarcinoma cells were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The expression of B-cell lymphoma 2 protein (Bcl-2), Bcl-2 associated X protein (Bax), caspase 9, and caspase 3 were examined by Western blot, to elucidate the possible molecular mechanisms involved. RESULTS: Our observations demonstrated that Dox-ZnO nanocomplexes could act as an efficient drug delivery system for importing Dox into SMMC-7721 cells, enhancing its potential chemotherapy efficiency by increasing the intracellular concentration of Dox. With the addition of ultraviolet (UV) illumination, the ZnO nanomaterials showed excellent photodynamic therapeutic properties, attacking the cancer cells further. Thus the caspase-dependent apoptosis was synergistically induced, resulting in distinct improvement in anticancer activity. CONCLUSION: The Dox-ZnO nanocomplex presents a promising multimodal agent for comprehensive cancer treatment.

Inclusion complexes of phosphorylated daidzein derivatives with ß-cyclodextrin: Preparation and inclusion behavior study.

In the present work the feasibility of ß-cyclodextrin in complexation was explored, as a tool for improving the solubility and biological ability of daidzein derivatives. A series of phosphorylated daidzein derivatives featuring different chain lengths were synthesized through a modified Atherton-Todd reaction and their inclusion complexes with ßCD were prepared by coprecipitation method. The inclusion complexation behavior was studied by fluorescence, UV, FT-IR, MS and (1)H NMR. The results showed that only phosphorylated daidzein derivative carrying small substituent group ((C(2)H(5)O)(2)PO) entered the cavity of ßCD and formed 1:1 inclusion complex. The formation constant was 175(mol/L)(-1).

Silencing of high mobility group A1 enhances gemcitabine chemosensitivity of lung adenocarcinoma cells.

BACKGROUND: The high mobility group A1 (HMGA1) proteins are architectural transcription factors found to be overexpressed in lung adenocarcinoma. Lentivirus-mediated RNA interference (RNAi) technology is a powerful tool for silencing endogenous or exogenous genes in human cancer cells. Our preliminary study shows that gemcitabine inhibits growth of the human lung cancer cell line SPCA-1 and induces apoptosis, and this effect might link with down-regulation of HMGA1 expression. This study aimed to investigate the chemosensitivity change of the lung adenocarcinoma cells SPCA-1 after HMGA1 inhibition by lentivirus-mediated RNAi. METHODS: We studied a highly malignant lung adenocarcinoma cell line (SPCA-1 cells). Lentiviral short-hairpin RNA (shHMGA1) expression vectors targeting HMGA1 were used for generation of lentiviral particles. After being transfected into the lung adenocarcinoma cell line SPCA-1, the expression of HMGA1 was determined by retrotranscriptase polymerase chain reaction (RT-PCR) and Western blotting. The effect of gemcitabine on proliferation of positive and negative cells was observed by methyl thiazolyl tetrazolium (MTT) assay and clonogenic survival assay. Apoptosis was observed by flow cytometery. Chemosensitivity to gemcitabine was determined by IC50 analysis. Caspase activity was quantitated by a caspase colorimetric protease assay kit. RESULTS: HMGA1-siRNA silenced its target mRNA specifically and effectively in SPCA-1 cells. The apoptotic rates of the scramble control group were (7.43 ± 0.21)%, (11.00 ± 0.20)%, and (14.93 ± 0.31)%, and the apoptotic rates in the silenced group were (9.53 ± 0.42)%, (16.67 ± 0.45)%, and (25.40 ± 0.79)% under exposure to 0.05, 0.5 and 5.0 µg/ml of gemcitabine (P < 0.05). The IC(50) of the silenced group was (0.309 ± 0.003) µg/ml which was significantly lower than in the scramble control group, (0.653 ± 0.003) µg/ml (P < 0.05). It reduced cancer cell proliferation and increased apoptotic cell death after being treated with gemcitabine compared with the scramble control group. HMGA1 silencing resulted in reduction in the phosphorylation of Akt, and promoted the activation of caspases 3, 8 and 9 upon exposure to gemcitabine. CONCLUSIONS: Lentivirus-mediated RNA interference of HMGA1 enhanced chemosensitivity to gemcitabine in lung adenocarcinoma cells. The mechanism may be associated with the PI-3K/Akt signal pathway. HMGA1 may represent a novel therapeutic target in lung cancer.

MDR reversal activity of bromotetrandrine in vitro and in vivo.

The present study was aimed to evaluate the MDR reversal activity of bromotetrandrine (BrTet) in vitro and in vivo. The inhibitory effects of adriamycin (ADM) used alone or in combination with BrTet or Tet on the proliferation of K562 and K562/A02 cells were evaluated by MTT assay. The ADM accumulation and the protein levels of P-glycoprotein (P-gp) were detected by flow cytometry. The mRNA levels of P-gp were determined by RT-PCR. The in vivo effect of BrTet and Tet was investigated by using nude mice grafted with sensitive human leukemia cell line K562 and MDR cell line K562/A02. The results showed that BrTet at 0.25, 0.5 and 1 micromol/L reversed the resistance to ADM in MDR K562/A02 cells in a dose-dependent manner. Flow cytometry suggested that BrTet significantly increased the intracellular accumulation of ADM in K562/A02 cells in a dose-dependent manner. BrTet also inhibited the overexpression of P-gp in K562/A02 cells, and down-regulated mdr1 expression. In nude mice bearing K562 xenografts on the left flank and K562/A02 xenografts on the right flank, intraperitoneal injection of 10 mg/kg BrTet significantly enhanced the antitumor activity of ADM against K562/A02 xenografts with inhibitory rates of 26.1%, while ADM alone inhibited the growth of K562/A02 xenografts only by 5.8%. No enhancement effect by BrTet was seen in K562 xenografts. It is concluded that BrTet shows significant MDR reversal activity in vitro and in vivo. Its activity may be related to the inhibition of P-gp overexpression and the increase intracellular accumulation of anticancer drugs. BrTet may be a promising-MDR modulator for eventual assessment in the clinic.

Effects of tetrandrine on apoptosis and radiosensitivity of nasopharyngeal carcinoma cell line CNE.

Tetrandrine is known to exert antitumor effect, however, little is known about its effect on nasopharyngeal carcinoma cells. In this study, we tested tetrandrine-induced apoptosis and radiosensitivity in nasopharyngeal carcinoma cell line CNE and investigated the possible mechanisms. Using flow cytometry and DNA electrophoresis, we found that tetrandrine could induce cell apoptosis. Further, it was shown that the level of Bcl-2 mRNA decreased and Bax mRNA increased after addition of tetrandrine by using reverse transcription-polymerase chain reaction. X-ray-induced G2 arrest was abrogated by treatment with tetrandrine, as detected by flow cytometry and mitotic index. The accumulation of cyclinB1 protein and the suppression of Cdc2 tyrosine-15 and Cdc25C serine-216 phosphorylation were detected in irradiated cells treated with tetrandrine using Western blot analysis. Taken together, these results show that tetrandrine can induce apoptosis and abrogate radiation-induced G2 arrest in CNE cells.

Tetrandrine: a potent abrogator of G2 checkpoint function in tumor cells and its mechanism.

OBJECTIVE: To assess the ability of tetrandrine (Tet) to enhance the sensitivity to irradiation and its mechanism in cell lines of human breast cancer p53-mutant MCF-7/ADR, p53-wild-type MCF-7 and human colon carcinoma p53-mutant HT-29 as well as in C26 colorectal carcinoma-bearing BALB/c mice. METHODS: MCF-7/ADR, HT-29 and MCF-7 cells were exposed to irradiation in the absence or presence of tetrandrine. The effect of Tet on the cytotoxicity of X-irradiation in these three cells was determined and the effect of tetrandrine on cell cycle arrest induced by irradiation in its absence or presence was studied by flow cytometry. Moreover, mitotic index measurement determined mitosis of cells to enter mitosis. Western blotting was employed to detect cyclin B 1 and Cdc2 proteins in extracts from irradiated or non-irradiated cells of MCF-7/ADR, HT-29 and MCF-7 treated with tetrandrine at various concentrations. Tumor growth delay assay was conducted to determine the radio-sensitization of tetrandrine in vivo. RESULTS: Clonogenic assay showed that tetrandrine markedly enhanced the lethal effect of X-rays on p53-mutant MCF-7/ADR and HT-29 cells and the sensitization enhancement ratio (SER) of tetrandrine was 1.51 and 1.63, but its SER was only 1.1 in p53-wt MCF-7 cells. Irradiated p53-mutant MCF-7/ADR and HT-29 cells were only arrested in G2/M phase while MCF-7 cells were arrested in G1 and G2/M phases. Radiation-induced G2 phase arrests were abrogated by tetrandrine in a concentration-dependent manner in MCF-7/ADR and HT-29 cells, whereas redistribution within MCF-7 cell cycle changed slightly. The proportion of cells in M phase increased from 1.3% to 14.7% in MCF-7/ADR cells, and from 1.5% to 13.2% in HT-29 cells, but 2.4% to 7.1% in MCF-7 cells. Furthermore, the levels of cyclin B 1 and Cdc2 expression decreased after X-irradiation in MCF-7/ADR and HT-29 cells, and the mitotic index was also lower. Tet could reverse the decrease and induce the irradiated cells to enter mitosis (M phase). Endosomatic experiment showed that tetrandrine caused tumor growth delay in irradiated mice. CONCLUSION: Tetrandrine boosts the cell killing activity of irradiation both in vitro and in vivo. Tetrandrine is a potent abrogator for G2 checkpoint control and can sensitize the cells to radiation.