Poor Prognostic Biomarker KIAA1522 Is Associated with Immune Infiltrates in Hepatocellular Carcinoma.

Background: The prognosis is poor for hepatocellular carcinoma (HCC), a tumor and cancer associated with inflammation that is common. New data showed that significant levels of KIAA1522 were expressed in HCC tissues and cell lines, suggesting that KIAA1522 may be a highly useful prognostic marker for HCC. However, its biochemical processes and impacts on the immune system go deeper. Objective: To verify the significance of KIAA1522 in HCC and investigate its related carcinogenic mechanisms. Methods: Studies examining the relationship between KIAA1522 expression and clinical-pathologic characteristics in HCC have been checked in the Cancer Genome Atlas (TCGA) database. A receiver operating characteristic (ROC) curve was used to assess the diagnostic efficacy of KIAA1522 in HCC. Western blot analysis was used to find the presence of the KIAA1522 protein in the tumor and paraneoplastic tissues of eight randomly chosen HCC patients. The GSVA program in R language was used to evaluate the relationship between KIAA1522 and immune cell infiltration in HCC. We searched the Search Tool for the Retrieval of Interacting Genes (STRING) database for interacting proteins connected to the expression of KIAA1522. Pathways were involved in the enrichment analysis of KIAA1522 to anticipate potential mechanisms through which KIAA1522 may affect immunological infiltration. Results: Our study found that KIAA1522 was commonly elevated in HCC tumor tissues and that it also signaled a bad outcome. We found an inverse link between KIAA1522 and cytotoxic cells and an inverse relationship between KIAA1522 and Th2 cell infiltration. In STRING analysis, the top 5 coexpressed proteins of KIAA1522 were BAIAP2, NCK2, TSNAXIP1, POGK, and KLHL31. The effect of KIAA1522 on HCC may entail cell cycle alteration, an immunological response, and suppression of the PPAR signaling pathway. Conclusion: High expression of KIAA1522 was linked to HCC immune cell infiltration, disease progression, and a poor prognosis.

Identification of Diagnostic Markers in Infantile Hemangiomas.

Background: Infantile Hemangiomas (IHs) are common benign vascular tumors of infancy that may have serious consequences. The research on diagnostic markers for IHs is scarce. Methods: The "limma" R package was applied to identify differentially expressed genes (DEGs) in developing IHs. Plugin ClueGO in Cytoscape software performed functional enrichment of DEGs. The Search Tool for Retrieving Interacting Genes (STRING) database was utilized to construct the PPI network. The least absolute shrinkage and selection operator (LASSO) regression model and support vector machine recursive feature elimination (SVM-RFE) analysis were used to identify diagnostic genes for IHs. The receiver operating characteristic (ROC) curve evaluated diagnostic genes' discriminatory ability. Single-gene based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) was conducted by Gene Set Enrichment Analysis (GSEA). The chemicals related to the diagnostic genes were excavated by the Comparative Toxicogenomics Database (CTD). Finally, the online website Network Analyst was used to predict the transcription factors targeting the diagnostic genes. Results: A total of 205 DEGs were singled out from IHs samples of 6-, 12-, and 24-month-old infants. These genes principally participated in vasculogenesis and development-related, endothelial cell-related biological processes. Then we mined 127 interacting proteins and created a network with 127 nodes and 251 edges. Furthermore, LASSO and SVM-RRF algorithms identified five diagnostic genes, namely, TMEM2, GUCY1A2, ISL1, WARS, and STEAP4. ROC curve analysis results indicated that the diagnostic genes had a powerful ability to distinguish IHs samples from normal samples. Next, the results of GSEA for a single gene illustrated that all five diagnostic genes inhibited the "valine, leucine, and isoleucine degradation" pathway in the development of IHs. WARS, TMEM2, and STEAP4 activated the "blood vessel development" and "vasculature development" in IHs. Subsequently, inhibitors targeting TMEM2, GUCY1A2, ISL1, and STEAP4 were mined. Finally, 14 transcription factors regulating GUCY1A2, 14 transcription factors regulating STEAP4, and 26 transcription factors regulating ISL1 were predicted. Conclusion: This study identified five diagnostic markers for IHs and further explored the mechanisms and targeting drugs, providing a basis for diagnosing and treating IHs.

Downregulation of MCF2L Promoted the Ferroptosis of Hepatocellular Carcinoma Cells through PI3K/mTOR Pathway in a RhoA/Rac1 Dependent Manner.

Methods and Results: The levels of MCF2L were detected by PCR and western blotting assay. The effect of MCF2L on ferroptosis was confirmed by MTT, colony formation assay, Brdu, in vivo animal experiment, and the content of Iron, GSH, ROS, and MDA. The underlying mechanisms were explored by PCR, western blotting, and affinity precipitation assay. Our findings demonstrated that MCF2L is remarkedly upregulated in HCC tissues, and sorafenib can induce the levels of MCF2L, suggesting that MCF2L might function in sorafenib resistance of HCC. Further analysis showed that downregulation of MCF2L enhances HCC cell death induced by sorafenib, and ferroptosis inhibitor can reverse this process. Subsequent experiments showed that downregulation of MCF2L elevates the content of Iron, ROS, and MDA, which are all indicators of ferroptosis. Finally, mechanism analysis showed that MCF2L regulates the PI3K/AKT pathway in a RhoA/Rac1 dependent manner. Conclusions: Our study showed that targeting MCF2L may be a hopeful method to overcome sorafenib-resistance through inducing ferroptosis in HCC.

Emergence and characterization of a putative novel human adenovirus recombinant HAdV-C104 causing pneumonia in Southern China.

In 2017, a survey of the molecular epidemiology of human adenovirus (HAdV) infections in Southern China based on hexon and fiber genotype demonstrated that the most prevalent genotypes of HAdV were HAdV-3 (n = 62), HAdV-2 (n = 21), and HAdV-7 (n = 16). In addition, two patients were co-infected with two genotypes of HAdV. Interestingly, a novel human adenovirus C recombinant genotype strain was isolated from one of the pneumonia patients in this survey. Phylogenetic, recombination, and proteotyping analysis showed that this novel pathogen originated from the recombination of parental viruses harboring the HAdV-1 penton and hexon gene, and the HAdV-2 fiber gene. It was named 'P1H1F2' and was assigned as HAdV-C104 based on the nomenclature protocol of using three major capsid proteins for characterization. Subsequent in vitro experiments demonstrated that HAdV-C104 had comparable proliferation capacity to HAdV-1, HAdV-2, and another recombination genotype P1H2F2. In addition, the HAdV-C104 infected patient was diagnosed with pneumonia and recovered after antiviral therapy. This report strengthens the hypothesis of recombination as a major pathway for the molecular evolution of HAdV-C species.

Background-Free Chromatographic Detection of Sepsis Biomarker in Clinical Human Serum through Near-Infrared to Near-Infrared Upconversion Immunolabeling.

Luminescence nanomaterial-based lateral flow assay (LFA) is promising for point-of-care tests. However, the detection sensitivity and accuracy are often affected by the interferences of autofluorescence and photon scattering from nitrocellulose membrane and colored plasma. Here, we describe a near-infrared to near-infrared upconversion nanoparticle (UCNP) immunolabeled LFA for background-free chromatographic detection of sepsis biomarker procalcitonin (PCT) in clinical human plasma. This upconversion immunolabeling enables both light excitation (at ∼980 nm) and anti-Stokes emission (at 800 nm) to be adopted within the first biological window (700-1000 nm), which eliminates background autofluorescence as well as photon scattering interferences, empowering a high-sensitivity detection without complicated procedures. After optimization, the described assay presented a limit of detection reaching down to 0.03 ng/mL, lower than the normal level (0.05 ng/mL), while having a detection range of 0.03-50 ng/mL that covers the clinical PCT level of interest (0.5-10 ng/mL). The assay recoveries in human serum samples were evaluated to be about 95-110%, whereas the inter- and intra-assay coefficient variations were both determined to be below 15%. Importantly, measured PCT concentrations in clinical samples are in good correlation with that of the electrochemiluminescence immunoassay (Roche) widely applied in large clinical settings. This near-infrared to near-infrared upconversion immunolabeling approach has direct implications for ultrasensitive and background-free point-of-care detection of other serum biomarkers in resource-limited clinical settings.

Detection of COVID-19: A review of the current literature and future perspectives.

The rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to the coronavirus disease 2019 (COVID-19) worldwide pandemic. This unprecedented situation has garnered worldwide attention. An effective strategy for controlling the COVID-19 pandemic is to develop highly accurate methods for the rapid identification and isolation of SARS-CoV-2 infected patients. Many companies and institutes are therefore striving to develop effective methods for the rapid detection of SARS-CoV-2 ribonucleic acid (RNA), antibodies, antigens, and the virus. In this review, we summarize the structure of the SARS-CoV-2 virus, its genome and gene expression characteristics, and the current progression of SARS-CoV-2 RNA, antibodies, antigens, and virus detection. Further, we discuss the reasons for the observed false-negative and false-positive RNA and antibody detection results in practical clinical applications. Finally, we provide a review of the biosensors which hold promising potential for point-of-care detection of COVID-19 patients. This review thereby provides general guidelines for both scientists in the biosensing research community and for those in the biosensor industry to develop a highly sensitive and accurate point-of-care COVID-19 detection system, which would be of enormous benefit for controlling the current COVID-19 pandemic.

Dysregulated NF-κB signal promotes the hub gene PCLAF expression to facilitate nasopharyngeal carcinoma proliferation and metastasis.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is common in Southern China. The molecular mechanism underlying NPC genesis and progression has been comprehensively investigated, but the key gene (s) or pathway (s) pertaining to NPC are unidentified. METHODS: We explored some key genes and pathways involved in NPC through using meta-analysis of deposited expression of microarray data of NPC. The expression of proliferating cell nuclear antigen clamp associated factor (PCLAF) was determined by real-time PCR and western blots. CCK-8 assay, colony formation assay, transwell migration assay, cell wound healing assay, cell cycle analysis and cell apoptosis were carried out to assess biological behaviors caused by downregulation and overexpression of PCLAF in vitro. CHIP was utilized to determine the direct upstream regulatory transcription factors of PCLAF. RESULTS: PCLAF was the key gene of NPC, which was significantly up-regulated in NPC cell line compared to the normal nasopharyngeal cell line. Additionally, in vitro assay has demonstrated the down-regulation and overexpression of PCLAF, resulted in significantly suppressed and enhanced NPC proliferation, metastasis and invasion respectively. Furthermore, the up-regulation of PCLAF in NPC is induced by direct binding of dysregulated NF-κB p50/RelB complex to the promoter of PCLAF. CONCLUSION: Our results offer a strategy for re-using the deposited data to find the key genes and pathways involved in pathogenesis of cancer. Our study has provided evidence of supporting the role of PCLAF in NPC genesis and progression.

Role of IQGAP3 in metastasis and epithelial-mesenchymal transition in human hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most lethal cancers worldwide owing to its high rates of metastasis and recurrence. The oncogene IQ motif-containing GTPase activating protein 3 (IQGAP3) is ubiquitously overexpressed in several human cancers, including liver, ovary, lung, large intestine, gastric, bone marrow, and breast malignancies and is involved in the invasion and metastasis of cancer cells. Therefore, we aimed to determine the biological role and molecular mechanism of IQGAP3 in HCC. METHODS: We used 120 archived clinical HCC samples, 9 snap-frozen HCC tumor tissues, and 4 normal liver tissues. Expression of IQGAP3 mRNA and protein in HCC cell lines (Hep3B, SMMC-7721, HCCC-9810, HepG2, BEL-7404, HCCLM3, QGY-7701, Huh7, and MHCC97H) and normal liver epithelial cells LO2 was examined by western blot, quantitative polymerase chain reaction, and immunohistochemistry. In addition, wound-healing and transwell matrix penetration assays were used to assess the migratory and invasive abilities of HCC cells, respectively. RESULTS: Expression of the IQGAP3 was robustly upregulated in HCC cells and tissues. High expression of IQGAP3 in HCC correlated with aggressive clinicopathological features and was an independent poor prognostic factor for overall survival. Furthermore, ectopic expression of IQGAP3 markedly enhanced HCC cell migration, invasion, and epithelial-to-mesenchymal transition (EMT) in vitro and promoted metastasis of orthotopic hepatic tumors in nude mice. Conversely, silencing endogenous IQGAP3 showed an opposite effect. Mechanistically, IQGAP3 promoted EMT and metastasis by activating TGF-ß signaling. CONCLUSIONS: IQGAP3 functions as an important regulator of metastasis and EMT by constitutively activating the TGF-ß signaling pathway in HCC. Our findings present new evidence of the role of IQGAP3 in EMT and metastasis, indicating its potential as a prognostic biomarker candidate and a therapeutic target against HCC.

Serine-arginine protein kinase 1 is associated with hepatocellular carcinoma progression and poor patient survival.

The pre-mRNA splicing regulator serine-arginine protein kinase 1 (SRPK1) promotes cancer development and various pathophysiological processes. However, the clinical relevance of SRPK1 in hepatocellular carcinoma (HCC) is not clear. This study investigates the expression and prognostic value of SRPK1 in HCC. We found that SRPK1 expression was significantly upregulated at the mRNA and protein level in all HCC cell lines or HCC tissue samples compared with the hepatic cell line or matched noncancerous tissue samples, respectively. Higher SRPK1 expression significantly correlated with clinical staging (p = 0.031), survival time (p = 0.004), and gender (p = 0.011) of HCC patients. Together, our study showed that SRPK1 is overexpressed in HCC and may be a promising indicator of prognosis for HCC patients.

MicroRNA-105 suppresses cell proliferation and inhibits PI3K/AKT signaling in human hepatocellular carcinoma.

A growing amount of evidence supports that microRNA (miRNA) dysregulation is involved in cancer progression by directly downregulating multiple targets. Elucidating the underlying mechanism of miRNA in carcinogenesis may improve diagnostic and therapeutic strategies for malignancy. In the current study, we found that miR-105 expression was markedly downregulated in both hepatocellular carcinoma (HCC) cell lines and clinical HCC tissues, compared with normal human hepatocyte and adjacent non-cancerous tissues, respectively. Ectopic miR-105 expression suppressed, whereas inhibiting miR-105 promoted the proliferation and tumorigenicity of HCC cells both in vitro and in vivo. Furthermore, we demonstrated that miR-105 could deactivated the phosphoinositide 3-kinase (PI3K)/AKT signaling pathway by downregulating insulin receptor substrate-1, 3-phosphoinositide-dependent protein kinase-1 and AKT1 directly, resulting in increasing cyclin-dependent kinase inhibitors 1A and 1B (p21(Cip1) and p27(Kip1)) and decreasing cyclin D1 expression in HCC. Therefore, our results suggest that miR-105 functions as a potential tumor suppressor by inhibiting the PI3K/AKT signaling pathway and might represent a potential therapeutic target for HCC patients.

[Effects of miR-106b expression on the proliferation of human hepatocellular carcinoma cells].

OBJECTIVE: To investigate the effects of miR-106b expression on the proliferation of human hepatocellular carcinoma cells. METHODS: Real-time PCR was used to detect the expression of miR-106b in human hepatocellular carcinoma (HCC) cell lines and normal liver epithelial THLE3 cells. Over-expression of miR-106b was transfected by miR-106b mimics, and inhibition of miR-106b expression was transfected by miR-106b inhibitors. The effects of miR-106b expression on the proliferation of HCC cells were detected by MTT, clone formation assay and anchorage-independent growth ability assay. Bromodeoxyuridine labeling and flow cytometry analysis were used to examine the effects of miR-106b expression on the cell cycle distribution of the HCC cells. RESULTS: Compared with that in the normal liver epithelial THLE3 cells, the relative expression of miR-106b in HepG2, QGY-7703, BEL-7402, MHCC97H, HCCC-9810, Hep3B, MHCC97L and Huh7 cell lines were 5.37 ± 0.35, 8.45 ± 0.75, 19.22 ± 1.74, 11.93 ± 1.26, 17.03 ± 0.97, 4.19 ± 0.67, 7.94 ± 1.35 and 3.82 ± 0.87, respectively (P < 0.05 for all). Three days after transfection, the miR-106b over-expression was accelerated, while miR-106b inhibitor suppressed the proliferation of HCC cells. The numbers of clones formed were (4.13 ± 0.75) and (3.78 ± 0.47) times higher than that of control cells, and (147.73 ± 15.56) and (138.87 ± 15.58) clones in diameter >1.0 mm were formed by miR-106b-overexpressing cells. When the miR-106b expression was inhibited in the HepG2 and QGY-7703 cells, the clone numbers were (0.18 ± 0.05) and (0.24 ± 0.07) times of that in the controls, and the numbers of clones formed were (23.29 ± 7.14) and (20.60 ± 8.07) (P < 0.05 for all). The positive rates of BrdU labeled miR-106b-overexpressing HepG2 and QGY-7703 cells were (51.89 ± 4.91) % and (54.74 ± 6.10) %, those of the miR-106b-inhibited cells were (6.48 ± 3.15) % and (7.52 ± 2.03) %, significantly different from that in the control cells (P < 0.05 for all). The S phases were dramatically increased from 29.93% and 31.04% to 56.76% and 57.22% in the miR-106b-overexpressing HepG2 and QGY-7703 cells, while they were 19.43% and 19.92% in the miR-106b-inhibited HepG2 and QGY-7703 cells. CONCLUSIONS: MiR-106b overexpression may promote the proliferation and metastasis of HCC cells. The mechanism of this effect may be related to promoting cells into S phase and inhibiting cell apoptosis.

MicroRNA-26b inhibits epithelial-mesenchymal transition in hepatocellular carcinoma by targeting USP9X.

BACKGROUND: Metastasis is responsible for the rapid recurrence and poor survival of malignancies. Epithelial-mesenchymal transition (EMT) has a critical role in metastasis. Increasing evidence indicates that EMT can be regulated by microRNAs (miRNAs). The aim of this study was to investigate the role of miR-26b in modulating epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma (HCC), as well as to identify its underlying mechanism of action. METHODS: The expression level of miR-26b was assessed in multiple HCC cell lines (HepG2, MHCC97H, Hep3B, MHCC97L, HCCC9810, BEL-7402, Huh7 and QGY-7703), as well as in liver tissue from patients with HCC. Follow-up studies examined the effects of a miR-26b mimic (increased expression) and a miR-26b inhibitor (decreased expression) on markers of EMT, wound healing and cell migration. The molecular target of miR-26b was also identified using a computer algorithm and confirmed experimentally. RESULTS: MiR-26b expression was decreased in HCC cell lines and was inversely correlated with the grade of HCC. Increased expression of miR-26b inhibited the migration and invasiveness of HCC cell lines, which was accompanied by decreased expression of the epithelial marker E-cadherin and increased expression of the mesenchymal marker vimentin, at both the mRNA and protein expression levels. A binding site for miR-26b was theoretically identified in the 3'UTR of USP9X. Further studies revealed that overexpression of miR-26b repressed the endogenous level of USP9X protein expression. Overexpression of miR-26b also repressed Smad4 expression, whereas its inhibition elevated Smad4 expression. CONCLUSIONS: Taken together, our results indicate that miR-26b were inhibited in HCC. In HCC cell lines, miR-26b targeted the 3'UTR of USP9X, which in turn affects EMT through Smad4 and the TGF-ß signaling pathway. Our analysis of clinical HCC samples verifies that miR-26b also targets USP9X expression to inhibit the EMT of hepatocytes. Thus, miR-26b may have some effects on the EMT of HCC cells.

miR-106b downregulates adenomatous polyposis coli and promotes cell proliferation in human hepatocellular carcinoma.

Aberrant activation of the Wnt/ß-catenin signal pathway is frequently observed in hepatocellular carcinoma (HCC). ß-Catenin is the major cellular effector of Wnt signaling and inactivation of adenomatous polyposis coli (APC) results in nuclear accumulation of ß-catenin. Therefore, it was speculated that APC inhibition could play important roles in activating the Wnt/ß-catenin pathway and in HCC progression. In this study, we report that miR-106b expression is markedly upregulated in hepatoma cells and hepatoma tissues compared with immortalized normal liver epithelial cells and normal hepatic tissues. Ectopic expression of miR-106b induces the proliferation and anchorage-independent growth of hepatoma cells, whereas inhibition of miR-106b reduced this effect. Furthermore, miR-106b upregulation in hepatoma cells modulated entry into the G(1)/S transitional phase by upregulating cyclin D1 and downregulating APC. Moreover, we demonstrated that miR-106b downregulates APC expression by directly targeting the 3'-untranslated region of APC messenger RNA. Taken together, our results suggest that miR-106b plays an important role in promoting the proliferation of human hepatoma cells and presents a novel mechanism of micro RNA-mediated direct suppression of APC expression in cancer cells.

[The role of Src kinase inhibitor ZD6474 on multi-drug resistant K562/A02 cells].

OBJECTIVE: To investigate the role of Src kinase inhibitor ZD6474 on the growth of multidrug-resistant K562/A02 cells and its regulatory mechanisms. METHODS: The possible mechanisms of drug-resistance were tested by Western blot. Proliferation assays and cell cycle distribution were analyzed by WST metric analysis. Western blot were used to investigate the mechanisms of antiproliferative activity induced by tyrosine kinase inhibitor ZD6474. The in vivo anti-tumor activity was evaluated in K562, K562/A02 xenografted nude mice by administration of ZD6474 (25 - 100 mg×kg(-1)×d(-1), PO). RESULTS: Compared with parental K562 cells, marked high levels of p-Src and Src expression were detected in K562/A02 cells. WST results showed that the IC(50) values of ZD6474 on K562 and K562/A02 after 48 hours incubation were (1.61 ± 0.07) µmol/L and (3.22 ± 0.21)µmol/L, respectively. ZD6474 caused an accumulation of cells in the G(0)/G(1) fraction and apoptosis by inhibiting the expressions of p-Src and Src kinase. Administration of ZD6474 produced a dose-dependent inhibition of tumor growth. 50 mg/kg ZD6474 produced the growth inhibition rates of 43.7% and 56.3%, respectively in K562 and K562/A02. CONCLUSION: Our results indicated that inhibiting Src kinase could induce K562/A02 cells apoptosis in vitro and in vivo.

IL-1 polymorphisms in children with peptic symptoms in South China.

BACKGROUND: Polymorphisms of IL-1 gene cluster are reported to be associated with histological changes and IL-1ß expression in the gastric mucosa in adults, especially in Helicobacter pylori-infected subjects. As H. pylori infecting adults and children own different virulence genotypes, the aim of this study was to investigate whether IL-1 polymorphisms are risk factors in young children in South China. MATERIALS AND METHODS: A total of 128 children with peptic symptoms were enrolled in this study. Polymorphisms of IL-1B-511 and IL-1B-31 were identified by dual fluorescence PCR. Variable number of tandem repeat region in IL-1RN was detected by conventional PCR and IL-1ß mRNA expression by real-time PCR ddCT assay. RESULTS: IL-1B-31T and IL-1B-511C were completely linked in this study. Significant differences of IL-1B-511/-31 genotypes were observed among different clinical outcomes (p = .001). The IL-1B-511TT/-31CC was mostly found in the moderate gastritis and the above (severe gastritis or gastric ulcer) groups, with percentage of 60.7%. While no association was observed between IL-1RN genotypes and the gastric mucosal histological changes (p = .128). Also no relationships were found between IL-1 polymorphisms and H. pylori infection or gastric mucosal IL-1ß mRNA expression level. CONCLUSION: Children with IL-1B-511TT/-31CC may have a risk to develop relatively severe gastric mucosal histological changes in South China.

Improved BC method of Compomat G4 for expression of BCs twice from whole blood in top and top bags.

The aim of this paper was to evaluate an improved buffy coat (BC) method of Compomat G4 for automated expression of BCs twice from whole blood (WB) in top and top (T&T) bags. WB was separated using hard spin centrifugation (2,988g, 10 min) into layers of blood components by specific gravity, and different components were subsequently expressed into satellite bags in the T&T system using the manual BC method, the conventional BC method of G4, and our improved BC method of G4. In the improved BC method, an accessorial device we have named a 'gravity press' was designed and installed on the top flat of G4 to produce gravitational pressure on the plasma bag so as to exclude air and some of plasma to the upper compartment of the slide after BCs were expressed for the first time. The residual BCs in the upper compartment were expressed a second time by extending the upper press once more. All of the pooled BCs were centrifuged by soft spin (402g, 10 min) and upper platelet-rich supernatant was manually expressed into a platelet container by the plasma extractor. In vitro studies of blood components and pooled platelet concentrates (PCs) revealed no significant differences in BC blood components and platelet recovery of pooled platelets (61 ± 9 vs. 60 ± 7%, n = 12, p > 0.05) between the improved BC method and the conventional BC method; all components met our specifications for blood products. We suggest that the new BC method for use of T&T bags may improve the collection of BCs.

[Effect of ZD6474 on the proliferation of imatinib-resistant K562 cells].

OBJECTIVE: To investigate the effect of tyrosine kinase inhibitor ZD6474 (Vandetanib) on the proliferative inhibition of K562 cells and its derived imatinib-resistant K562/G cells and its mechanism. METHODS: Imatinib-resistant K562/G cells were obtained by culturing cells in gradually increasing concentrations of imatinib. The changed factors related to drug-resistance were tested by Western blot. ZD6474 and imatinib affected K562/G and parental K562 cells proliferation were analyzed by WST assay. Flow cytometry was used to analyze cell cycle. Direct inhibition of Src activity by ZD6474 was measured by a colorimetric ELISA assay with recombinant human Src kinase. RESULTS: 10 µmol/L imatinib failed to inhibit K562/G cells proliferation or induce cell cycle arrest. Compared with that in parental K562 cells, there were marked high levels of p-Src and Src protein in K562/G cells. The expression of Bcl-2 and p-STAT3 also increased in K562/G cells. After 48 hours incubation, the IC(50) values of ZD6474 in K562 and K562/G cells were 1.61 µmol/L and 3.18 µmol/L, respectively. ZD6474 treatment caused accumulation of cells in the G(0)/G(1) fraction and cell apoptosis in K562 and K562/G cells. ZD6474 decreased the expression of p-Src and Src at post-transcriptional level. Moreover, ZD6474 increased the ratio of Bax/Bcl-2 and decreased the expression of p-STAT3 at the same concentration for inducing apoptosis. CONCLUSIONS: ZD6474 is effective in inhibiting the proliferation of imatinib-resistant K562/G cells and parental K562 cells, and induces their apoptasis by significant inhibition of Src kinase activity. Our study provides a reliable experimental basis for chronic myeloid leukemia treatment with ZD6474.

ZD6474 inhibits Src kinase leading to apoptosis of imatinib-resistant K562 cells.

ZD6474 is an orally available, small-molecule tyrosine kinase inhibitor. This study explores the effect of ZD6474 on imatinib-resistant K562 cell lines, which show markedly increased SRC family kinases (SFKs) activity. ZD6474 induces growth arrest and apoptosis of imatinib-resistant and parental K562 cells, as well as inhibition of Src activity and its downstream effectors, the anti-apoptotic Bcl-2 family. ZD6474 treatment also inhibits the activity of STAT3 and reactivation of its activity results in suppression of the anti-tumor effects of SFKs inhibitors. A single oral administration of ZD6474 produced dose-dependent inhibition of imatinib-resistant K562 cells xenograft tumors. These results suggest that clinical assessment of ZD6474 against imatinib-resistant CML is warranted.

Dynamic distribution and expression in vivo of the human interferon gamma gene delivered by adenoviral vector.

BACKGROUND: We previously found that r-hu-IFNgamma exerts a potent anti-tumor effect on human nasopharyngeal carcinoma xenografts in vivo. Considering the fact that the clinical use of recombinant IFNgamma is limited by its short half-life and systemic side effects, we developed a recombinant adenovirus, Ad-IFNgamma. METHODS: Dynamic distribution of the adenovirus vector and expression of IFNgamma were evaluated by Q-PCR and ELISA after intratumoral administration of Ad-IFNgamma into CNE-2 xenografts. RESULTS: Ad-IFNgamma DNA was mainly enriched in tumors where the Ad-IFNgamma DNA was injected (P < 0.05, compared to blood or parenchymal organs), as well as in livers (P < 0.05). Concentrations of Ad-IFNgamma DNA in other organs and blood were very low. Intratumoral Ad-IFNgamma DNA decreased sharply at high concentrations (9 x 10(5) copies/microg tissue DNA), and slowly at lower concentrations (1.7-2.9 x 10(5) copies/microg tissue DNA). IFNgamma was detected in the tumors and parenchymal organs. The concentration of IFNgamma was highest in the tumor (P < 0.05), followed by the liver and kidney (P < 0.05). High-level intratumoral expression of IFNgamma was maintained for at least 7 days, rapidly peaking on day 3 after injection of Ad-IFNgamma DNA. CONCLUSION: An IFNgamma gene delivered by an adenoviral vector achieved high and consistent intratumoral expression. Disseminated Ad-IFNgamma DNA and the transgene product were mainly enriched in the liver.