RESUMEN
AIM: To construct a novel liposomal drug delivery system co-modified with SP94 and BR2 ligands, encapsulating both the bitter ginseng derivative B21 and doxorubicin (DOX), to achieve superior anti-tumour efficacy and reduced toxic side effects. METHODS: Liposomes were prepared using an organic phase reaction method, with B21 encapsulated in the lipid phase and DOX in the aqueous phase. The liposomes were further modified with SP94 and BR2 peptides. The characterisations, cytotoxicity, and in vitro targeting effects were assessed through various methods including ultraviolet spectrophotometry, high-performance liquid chromatography, nano-size analysis, ultrafiltration centrifugation, dialysis, transmission electron microscopy, flow cytometry, Methylthiazolyldiphenyl-tetrazolium bromide assay, confocal laser scanning microscopy, transwell assay, and tumorsphere assay. RESULTS: SP94/BR2-B21/DOX-LP liposomes were spherical with an average diameter of 120.87 ± 1.00 nm, a polydispersity index (PDI) of 0.223 ± 0.006, and a surface charge of -23.1 ± 1.27 mV. The encapsulation efficiencies for B21 and DOX were greater than 85% and 97% (mg/mg), respectively. The results indicated that SP94/BR2-B21/DOX-LP exhibited enhanced targeting and cytotoxicity compared to single-ligand modified and unmodified liposomes, with the combined encapsulation of B21 and DOX showing synergistic anti-hepatocarcinogenic effects. CONCLUSION: SP94/BR2-B21/DOX-LP liposomes represent a promising targeted drug delivery system for hepatocellular carcinoma, offering improved membrane penetration, enhanced therapeutic efficacy, and reduced systemic toxicity.
RESUMEN
Insufficient ventricular unloading is a serious complication during veno-arterial extracorporeal membrane oxygenation (VA-ECMO) that has a crucial impact on patient outcomes. The existing conservative treatment options are limited, while mechanical decompression techniques are challenging and restricted in terms of their adoption and application. Two patients with cardiogenic shock experienced insufficient left ventricular unloading with no pulsatile contraction and aortic valve closure during VA-ECMO support. Gentle chest compression was applied to establish an active left ventricular drainage mechanism, which prevented the formation of intracardiac thrombi. No life-threatening complications or technical problems occurred. Therefore, gentle chest compression was established as an effective and safe method for treating insufficient left ventricular unloading in VA-ECMO patients.
RESUMEN
AIM: To construct a novel nano-carrier with dual ligands to achieve superior anti-tumour efficacy and lower toxic side effects. METHODS: Liposomes were prepared by thin film hydration method. Ultraviolet, high performance liquid chromatography, nano-size analyser, ultrafiltration centrifugation, dialysis, transmission electron microscope, flow cytometry, Cell Counting Kit-8, confocal laser scanning microscopy, transwell, and tumorsphere assay were used to study the characterisations, cytotoxicity, and in vitro targeting of dg-Bcan targeting peptide (BTP-7)/pHA-temozolomide (TMZ)/tetra(4-carboxyphenyl)porphyrin (TCPP)-Lip. RESULTS: BTP-7/pHA-TMZ/TCPP-Lip was a spheroid with a mean diameters of 143 ± 3.214 nm, a polydispersity index of 0.203 ± 0.025 and a surface charge of -22.8 ± 0.425 mV. The drug loadings (TMZ and TCPP) are 7.40 ± 0.23% and 2.05 ± 0.03% (mg/mg); and the encapsulation efficiencies are 81.43 ± 0.51% and 84.28 ± 1.64% (mg/mg). The results showed that BTP-7/pHA-TMZ/TCPP-Lip presented enhanced targeting and cytotoxicity. CONCLUSION: BTP-7/pHA-TMZ/TCPP-Lip can specifically target the tumour cells to achieve efficient drug delivery, and improve the anti-tumour efficacy and reduces the systemic toxicity.
Asunto(s)
Glioblastoma , Liposomas , Temozolomida , Glioblastoma/tratamiento farmacológico , Humanos , Línea Celular Tumoral , Temozolomida/farmacología , Temozolomida/administración & dosificación , Temozolomida/farmacocinética , Temozolomida/química , Porfirinas/química , Porfirinas/administración & dosificación , Porfirinas/farmacología , Sistemas de Liberación de Medicamentos , Neoplasias Encefálicas/tratamiento farmacológico , Péptidos/química , Péptidos/farmacologíaRESUMEN
Lung cancer is characterized by its high mortality and morbidity. A deep understanding of the molecular mechanisms of lung cancer tumorigenesis helps to develop novel lung cancer diagnostic and therapeutic strategies. However, the picture of the associated molecular landscape is not yet complete. As understood, chemokine-receptor interactions contribute much to lung cancer tumorigenesis, in which CCR10 also plays an important role. This study aimed to expand the knowledge of CCR10 in lung squamous cell carcinoma (LUSC) in the manner of molecular mechanism and biological functions. Using GEPIA database, the survival analysis between LUSC patients with high and low CCR10 expressions was performed, showing that CCR10 could be regarded as a risk factor for LUSC patients. Subsequently, CCR10 protein and mRNA expressions in LUSC were examined by qRT-PCR and western blot respectively. The results indicated that CCR10 was highly expressed in LUSC cells. The results of CCK-8, colony formation, and Transwell assays presented that CCL27, the ligand of CCR10, promoted proliferative, migratory, and invasive abilities of LUSC cells by activating CCR10. Also, the PI3K/AKT signaling pathway was verified as the involved pathway by western blot. Overall, it could be concluded that the CCL27-CCR10 regulatory axis can activate the PI3K/AKT pathway fostering the malignant features of LUSC cells.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas , Carcinoma de Células Escamosas , Neoplasias Pulmonares , Humanos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Carcinoma de Células Escamosas/genética , Carcinoma de Pulmón de Células no Pequeñas/genética , Neoplasias Pulmonares/genética , Carcinogénesis/genética , Proliferación Celular , Pulmón/metabolismo , Regulación Neoplásica de la Expresión Génica , Receptores CCR10/genética , Receptores CCR10/metabolismo , Quimiocina CCL27/genética , Quimiocina CCL27/metabolismoRESUMEN
Lung adenocarcinoma (LUAD) is one of the most commonly diagnosed subtypes of lung cancer, and one of the deadliest cancers. Tetratricopeptide repeat domain 9A (TTC9) is upregulated and has played an oncogenic role in some malignant tumors. However, the expression and role of TTC9 has not yet been elucidated in LUAD. Here, we investigated the expression profiles, biological functions and potential molecular mechanism of the TTC9 gene in LUAD. TTC9 expression was significantly overexpressed in LUAD tissues compared with that in normal lung tissues. TTC9 expression was closely correlated with gender, lymph node metastasis, and survival status in the TCGA-LUAD cohort. Subsequent cellular function assays demonstrated that knockdown of TTC9 promoted PC9 cell apoptosis and inhibited cell proliferation, migration and invasion, leading to cell cycle arrest in G2 phase. Moreover, inhibition of TTC9 suppressed the tumorigenicity of PC9 cells in nude mice. TTC9 might serve as oncogene in LUAD through cancer-related signaling pathways including p38 MAPK pathway. The expression of TTC9 gene might be modulated by DNA copy number variant and DNA methylation. TTC9 was significantly associated with tumor immune infiltration patterns. Accordingly, TTC9 may be a novel therapeutic target for the treatment of LUAD.
RESUMEN
OBJECTIVE: This study attempted to investigate the mechanism of miR-204-5p and its downstream gene in regulating bio-functions of esophageal cancer (EC). METHODS: Bioinformatics analysis was performed to select the mature miRNAs, mRNAs, and clinical data of EC. The miRNA-mRNA regulatory axis was predicted through bioinformatics and used Dual-luciferase analysis to verify the interaction between miR-204-5p and APLN. qRT-PCR was applied to analyze expression of miR-204-5p and APLN mRNA. Western blot was utilized to detect APLN protein expression. Functional assays like CCK-8, wound healing, Transwell, and stem cell sphere formation assays were launched to confirm proliferative, migratory, invasive and stemness of cells in different treatment groups. RESULTS: MiR-204-5p was lowly expressed while its target gene APLN was highly expressed in tumor tissues. Besides, miR-204-5p overexpression hindered proliferation, invasion, migration, and stemness of EC cells. Additionally, dual-luciferase assay verified the interaction of miR-204-5p and APLN. MiR-204-5p could downregulate APLN level and its overexpression reduced the effect of APLN on EC cell functions. CONCLUSION: Dysregulation of miR-204-5p/APLN axis was linked with malignant progression of EC. MiR-204-5p/APLN may be an underlying candidate for the design of anticarcinogens.
Asunto(s)
Apelina , Neoplasias Esofágicas , MicroARNs , Humanos , Apelina/genética , Apelina/metabolismo , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patología , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , ARN MensajeroRESUMEN
Excision repair crosscomplementation group 6 like (ERCC6L) has been reported to be upregulated in a variety of malignant tumors and plays a critical oncogenic role. However, the role and molecular mechanism of ERCC6L in lung adenocarcinoma (LUAD) remain unclear, and were therefore investigated in the present study. Clinical data of patients with LUAD were obtained and bioinformatics analysis was performed to investigate the expression characteristics, prognostic value, and biological function of ERCC6L. In addition, cell function experiments were performed to detect the effect of ERCC6L silencing on the biological behavior of LUAD cells. The results revealed that ERCC6L expression was significantly higher in LUAD tissues vs. normal lung tissues and closely associated with nodal invasion, advanced clinical stage and survival in LUAD. Overexpression of ERCC6L was an independent prognostic biomarker of overall survival, progressionfree interval, and diseasespecific survival in patients with LUAD. DNA amplification and low methylation levels of ERCC6L suggested regulation at both the genetic and epigenetic levels. The most significant positive genes coexpressed with ERCC6L were mainly enriched in the cell cycle signaling pathway. The major functions of ERCC6L in LUAD cells were positively correlated with the cell cycle, DNA damage, DNA repair, proliferation, invasion and epithelialmesenchymal transition (EMT). Knockdown of ERCC6L inhibited the proliferative, migratory and invasive abilities of A549 and PC9 cells. It also promoted cell apoptosis, and led to cell cycle arrest in the S phase. ERCC6L may regulate the EMT process through the Wnt/ßcatenin and Wnt/Notch 3 signaling pathways, thus regulating the tumorigenesis and progression of LUAD. The overexpression of ERCC6L may be a biological indicator for the diagnosis and prognosis of LUAD. ERCC6L may be a novel molecular target for the treatment of lung cancer.
Asunto(s)
Adenocarcinoma del Pulmón , ADN Helicasas , Neoplasias Pulmonares , Adenocarcinoma del Pulmón/patología , Proliferación Celular/genética , ADN , ADN Helicasas/genética , Humanos , Neoplasias Pulmonares/patología , Fenotipo , PronósticoRESUMEN
BACKGROUND: Kinesin family member 18B (KIF18B) is a member of the kinesin-8 superfamily, and functions as an oncogene in human cancers. However, its expression profile and role in lung adenocarcinoma (LUAD) remain unclear. MATERIALS AND METHODS: We examined the expression profile of KIF18B using quantitative real-time reverse transcription polymerase chain reaction and immunohistochemistry in fresh clinical samples. Using data downloaded from the Cancer Genome Atlas database and Gene Expression Omnibus, we explored the clinical significance of KIF18B, potential mechanisms of its dysregulation and its underlying biological function in LUAD. RESULTS: KIF18B was significantly over-expressed in LUAD tissues relative to normal tissues. High KIF18B expression was associated with smoking history, positive nodal invasion, advanced clinical stage, death status and poorer prognosis. Cox regression analyses revealed that KIF18B overexpression was an independent prognostic biomarker for poor overall survival (OS) and recurrence-free survival in LUAD. In addition, KIF18B mutation was observed in 2.2% of LUAD cases. DNA copy number variation was correlated with upregulated expression of KIF18B in LUAD tissues and cell lines. The methylation level of some KIF18B DNA CpG sites was negatively associated with its mRNA expression. KIF18B was predictively targeted by miR-125a-5p, which was downregulated in LUAD tissues, inversely correlated with KIF18B mRNA expression and significantly associated with poor OS. Furthermore, gene set enrichment analysis revealed that genes positively co-expressed with KIF18B were mainly enriched in cell cycle signaling pathways. CONCLUSION: Our results indicate that KIF18B is a promising prognostic biomarker for LUAD. DNA amplification, hypomethylation as well as miR-125a-5p downregulation may be involved in the mechanism of KIF18B dysregulation in LUAD. KIF18B might function as a novel oncogene through cell cycle regulation pathways in LUAD.
RESUMEN
OBJECTIVE: To evaluate the effectiveness of autologous pericardium ring in tricuspid annuloplasty surgery for the treatment of tricuspid regurgitation (TR). METHODS: From December 2010 to December 2012, a total of 107 patients with secondary TR underwent tricuspid annuloplasty. The patients were divided into three groups: autologous pericardium ring group (n = 38), Edwards-MC3 ring group (n = 35), and DeVega group (n = 34). The patients were followed-up for two years. The survival rates and free from hospital readmission rates were measured and analyzed. The patients also received transthoracic echocardiography (TTE) in order to obtain TR regurgitant jet area to right atrial area (STR/STA), diastolic tricuspid annuloplasty diameter (DTAD), right atrial diameter (RAD), and right ventricular diameter (RVD). RESULTS: One patient from DeVega group and one patient from autologous pericardium ring died from low cardiac output syndrome during the perioperative period. In the two-year follow-up period, each group has one instance of death for unclear reasons. One month after operation, the STR/STA, DTAD, RAD, and RVD values in all groups were significantly lower than the pre-operation values (P < 0.05). During the two year follow-up period, DTAD values of patients from DeVega group increased significantly as compared to the values at one month post operation (P<0.05), which is different from the other two groups in which DTAD values remained stable (P>0.05). In both pericardium ring group and Edwards-MC3 group, STR/SRA, remained stable (P>0.05) during the follow-up period, whereas STR/SRA of the DeVega group had showed a tendency of increase (although statistically insignificant, P>0.05). There was no significant difference in the survival rates among three study groups (P > 0.05), but the rate of free from hospital readmission in the DeVega group was significantly lower than those in the other two groups (P < 0.05) during the two-year follow-up period. CONCLUSIONS: Autologous pericardium tissue based ring annuloplasty demonstrated remarkable clinical utility for treating tricuspid regurgitation. It shows similar beneficial results to Edwards-MC3 annuloplasty within a short-term follow-up period, and outperforms the widely used DeVega annuloplasty. Autologous pericardium tissue annuloplasty represents a promising technique for tricuspid annuloplasty and holds great potential for treating tricuspid valve dysfunctions.
Asunto(s)
Anuloplastia de la Válvula Cardíaca/métodos , Pericardio/trasplante , Insuficiencia de la Válvula Tricúspide/diagnóstico por imagen , Insuficiencia de la Válvula Tricúspide/cirugía , Válvula Tricúspide/cirugía , Adulto , Ecocardiografía , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Readmisión del Paciente , Estudios Retrospectivos , Tasa de Supervivencia , Trasplante Autólogo , Resultado del TratamientoRESUMEN
Kinesin family member 18A (KIF18A), as a member of the kinesin superfamily, is significantly overexpressed and abnormally functions in various human cancers. But, its expression profiling in the lung adenocarcinoma (LUAD) remains unclear. In the present work, using the data derived from the Cancer Genome Atlas (TCGA), we assessed the expression pattern and prognostic value of KIF18A in LUAD. In addition, we analyzed the underlying mechanism of its gene dysregulation. Experimental and bioinformatic analysis results showed that KIF18A expression was dramatically increased in LUAD tissues compared with the normal counterparts. Moreover, the patients with high KIF18A expression had significantly poorer overall survival (OS) and recurrence-free survival (RFS). Univariate and multivariate analyses indicated that increased KIF18A expression was independently associated with unfavorable OS and RFS. In addition, by analyzing deep sequencing data from TCGA-LUAD, we found that KIF18A mutation was detected in 2.6% of LUAD cases, and increased KIF18A expression was associated with genetic amplification rather than DNA methylation. Moreover, gene co-expression network analysis revealed that a total of 339 KIF18A co-expressed genes were detected and enriched in several tumor-related pathways, especially cell cycle. Knockdown of KIF18A significantly inhibited cell proliferation in vitro and in vivo. Furthermore, silencing KIF18A induced LUAD cells apoptosis and arrested the cell cycle in the G2/M phase. KIF18A promotes cell proliferation, inhibits apoptosis, and is a valuable prognostic predictor and potential therapeutic target for the patients with LUAD. © 2019 IUBMB Life, 2019.
Asunto(s)
Adenocarcinoma del Pulmón/patología , Apoptosis , Biomarcadores de Tumor/metabolismo , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Cinesinas/metabolismo , Neoplasias Pulmonares/patología , Adenocarcinoma del Pulmón/genética , Adenocarcinoma del Pulmón/metabolismo , Anciano , Animales , Biomarcadores de Tumor/genética , Ciclo Celular , Metilación de ADN , Femenino , Perfilación de la Expresión Génica , Humanos , Cinesinas/genética , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Persona de Mediana Edad , Pronóstico , Tasa de Supervivencia , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
We developed a new minimally invasive thoracoscopic technique of extended thymectomy for myasthenia gravis by combining a subxiphoid incision with dual costal margin incisions. In our experience of 31 consecutive cases, this procedure provides a good operative view in the neck region and makes verification of the bilateral phrenic nerves easy. All the patients recovered smoothly with less trauma, less bleeding, less complication, and good cosmetic results. This modified transsubxiphoid approach is a satisfactory procedure for performing extended thymectomy in patients with myasthenia gravis.
Asunto(s)
Miastenia Gravis/cirugía , Cirugía Torácica Asistida por Video/métodos , Timectomía/métodos , Adulto , Pérdida de Sangre Quirúrgica , Drenaje , Femenino , Humanos , Tiempo de Internación , Masculino , Miastenia Gravis/diagnóstico , Complicaciones Posoperatorias/etiología , Factores de Riesgo , Cirugía Torácica Asistida por Video/efectos adversos , Timectomía/efectos adversos , Factores de Tiempo , Resultado del TratamientoRESUMEN
We report a rare case of dextroversion accompanied with atrial septal defect (ASD), persistent left superior vena cava with absent right superior vena cava in a four-year-old male. A polytetrafluoroethylene (PTFE) graft as an extracardiac conduit was used to connect the persistent left superior vena cava (PLSVC) with the right atrial appendage.
Asunto(s)
Apéndice Atrial/anomalías , Apéndice Atrial/cirugía , Implantación de Prótesis Vascular/métodos , Procedimientos Quirúrgicos Cardiovasculares/métodos , Dextrocardia/cirugía , Defectos del Tabique Interatrial/cirugía , Vena Cava Superior/anomalías , Vena Cava Superior/cirugía , Prótesis Vascular , Preescolar , Humanos , Masculino , Politetrafluoroetileno , Resultado del TratamientoRESUMEN
BACKGROUND: The CC chemokine receptor 9 (CCR9) plays an important role in tumorigenesis and metastasis in various cancers. Our previous studies have shown the aberrant expression of CCR9 in non-small cell lung cancer (NSCLC) cell lines, revealing that the CCR9-CCL25 axis modulates cell migration and invasion, and supports cancer cell survival by inhibiting apoptosis in vitro and in vivo. In the present study, we aimed to evaluate the expression and possible prognostic role of CCR9 in lung adenocarcinoma. METHODS: Immunohistochemical analysis of CCR9 expression was performed on 144 lung adenocarcinoma tissues and 30 adjacent normal lung parenchymal tissues. We assessed the correlation of CCR9 expression with clinicopathological characteristics and the prognosis of lung adenocarcinoma. RESULTS: The expression of CCR9 was increased in lung adenocarcinoma tissue compared with normal lung tissue. Moreover, such an expression was positively correlated with tumor size (p = 0.032), lymph node metastasis (p = 0.002) and advanced TNM stage (p = 0.012). In addition, the patients with negative CCR9 expression exhibited a higher overall survival (OS) compared with those with positive CCR9 expression. Multivariate analysis showed that the CCR9 expression was an independent prognostic factor for the OS of patients with lung adenocarcinoma. CONCLUSIONS: We, for the first time, reported that CCR9 could be beneficial in predicting lymph node metastasis, and it might act as a novel prognostic biomarker for lung adenocarcinoma.
Asunto(s)
Adenocarcinoma/química , Biomarcadores de Tumor/análisis , Neoplasias Pulmonares/química , Receptores CCR/análisis , Adenocarcinoma/mortalidad , Adenocarcinoma/secundario , Adenocarcinoma del Pulmón , Adulto , Anciano , Distribución de Chi-Cuadrado , Femenino , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Neoplasias Pulmonares/mortalidad , Neoplasias Pulmonares/patología , Metástasis Linfática , Masculino , Persona de Mediana Edad , Análisis Multivariante , Estadificación de Neoplasias , Valor Predictivo de las Pruebas , Modelos de Riesgos Proporcionales , Factores de Riesgo , Factores de Tiempo , Carga Tumoral , Regulación hacia ArribaRESUMEN
CC chemokine receptor-9 (CCR9) is highly expressed in non-small cell lung cancer (NSCLC) tissues and cell lines. However, the biological functions and the signals elicited by the interaction between CCR9 and its natural ligand CCL25 in NSCLC are unknown. Here, we selectively depleted CCR9 and inhibited CCR9-CCL25 interaction in NSCLC cells using small recombinant lentivirus-mediated miRNA, and investigated the tumorigenic effects in vitro and in vivo. Compromised CCR9-CCL25 interaction promoted apoptosis in NSCLC cells by activating phosphoinositide 3-kinase (PI3K)/Akt in vitro. In addition, we showed that CCR9-CCL25 interaction mediated the activation of the PI3K/Akt pathway in NSCLC cells, resulting in the up-regulation of anti-apoptotic proteins, as well as the down-regulation of apoptotic proteins in a PI3K-/Akt-dependent manner. These CCR9-CCL25-mediated effects were abrogated in the presence of a PI3K inhibitor (wortmannin 10 nM) or by inhibiting the CCR9-CCL25 interaction through CCR9 silencing, which also suggested that the biological function of CCR9-CCL25 was mainly regulated by PI3K. In vivo studies also demonstrated a significantly lower tumor burden in mice receiving CCR9-silence cells than those in mice receiving control cells. Together, these data suggested that CCR9-CCL25 interaction induced tumorigenesis of NSCLC cells and that this induction might be accomplished through the activation of the PI3K/Akt pathway. These findings may lead to a better understanding of the biological effects of CCR9-CCL25 interaction and provide clues for identifying novel therapeutic and preventive molecular markers for NSCLC.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/patología , Quimiocinas CC/metabolismo , Neoplasias Pulmonares/patología , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptores CCR/genética , Androstadienos/farmacología , Animales , Apoptosis/genética , Secuencia de Bases , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Línea Celular Tumoral/efectos de los fármacos , Quimiocinas CC/genética , Inhibidores Enzimáticos/farmacología , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Inhibidores de las Quinasa Fosfoinosítidos-3 , Receptores CCR/metabolismo , Wortmanina , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
CD59, belonging to membrane complement regulatory proteins (mCRPs), inhibits the cytolytic activity of complement and is overexpressed in many types of solid cancers. The aim of the present study was to detect the expression of CD59 in non-small cell lung cancer (NSCLC) and to investigate the relationship between decreased CD59 expression and tumorigenesis of NSCLC by transfecting recombinant retrovirus encoding shRNA targeting human CD59 into the human NSCLC cell line NCI-H157. CD59 expression in NSCLC was detected by immunocytochemistry (IHC). In the human NSCLC cell line NCI-H157, CD59 mRNA and protein expression suppressed with lentivirus-mediated RNAi was confirmed by using RT-PCR and western blotting, respectively. The proliferation and apoptosis of NCI-H157 cells was measured by using MTT assay and FACS. The resistance to complement cracking ability was detected by LDH assay. Caspase-3 expression in cells was assessed by IHC. Bcl-2 and Fas protein was determined by western blotting both in vitro and in vivo. CD59 is overexpressed in human NLCLC cancer. In NCI-H157 cells, lentivirus-mediated RNAi significantly reduced both CD59 mRNA and protein expression, which resulted in suppressing cell proliferation and increasing cell apoptosis. When incubated with fresh normal human serum (8%, v/v) for 1 h at 37ËC, the cell viability was decreased and cell apoptosis was increased in siCD59-infected NCI-H157 cells compared to siCD59-C-infected cells. Reduced CD59 expression led to increased expression of caspase-3 and Fas and decreased expression of Bcl-2. Furthermore, the nude mouse tumor graft weight was significantly decreased and survival rate was significantly increased in the siCD59 group. CD59 is overexpressed in human NLCLC. CD59 silencing in NSCLC cancer cells via retrovirus-mediated RNAi can enhance complement-mediated cell apoptosis, inhibiting the growth of NSCLC. CD59 may serve as a potential target for gene therapy in NSCLC.
Asunto(s)
Antígenos CD59/genética , Carcinoma de Pulmón de Células no Pequeñas/genética , Neoplasias Pulmonares/genética , ARN Mensajero/genética , Animales , Apoptosis/genética , Antígenos CD59/biosíntesis , Carcinoma de Pulmón de Células no Pequeñas/patología , Línea Celular Tumoral , Proliferación Celular , Supervivencia Celular/genética , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Pulmonares/patología , Ratones , Interferencia de ARNRESUMEN
BACKGROUND: Previously, we reported the preservation method of donor hearts in an empty beating status with mild hypothermic perfusion. To completely avoid cardiac arrest and myocardial ischemia, we performed the beating preservation technique from procurement of hearts to transplants and assessed its efficacy for long-term preservation and feasibility for heart transplantation. METHODS: Thirty-two swine donor hearts were preserved in beating status (group A, n = 8 pairs, perfused continuously with normothermic blood) or in static cold storage (group B, n = 8 pairs, stored in 4°C histidine-tryptophan-ketoglutarate solutions) for 8 hours. Then the donor hearts were implanted either in beating or static status. During transplantation, the incidence of arrhythmia, duration of anastomosis and cardiopulmonary bypass, and dosage of inotropic drugs were recorded. Hemodynamics of left ventricle and serum level of creatine kinase-MB were measured during transplantation. Myocardial ultrastructure was observed. RESULTS: Compared with group B, in group A the anastomotic time was significantly longer, the cardiopulmonary bypass time was significantly shorter, the cardiac output was larger, and the incidence of arrhythmia, dosage of cardiovascular-active drugs, and serum level of creatine kinase-MB were lower. After declamping for 2 hours and 3.5 hours, the left ventricular hemodynamics of group A was significantly better than that of group B. The myocardial ultrastructure of group A was superior to that of group B. CONCLUSIONS: Preservation of donor hearts in beating status with continuous, normothermic, blood perfusion is an effective approach for long-term preservation and is appropriate for heart transplantation.