RESUMEN
OBJECTIVES: Chromosome segregation 1-like (CSE1L) is abundant and strongly expressed in solid tumors. However, the expression and role of CSE1L in chronic myeloid leukemia(CML) remain largely unknown. MATERIALS AND METHODS: The relative expression levels of CSE1L in bone marrow granulocytes from patients with primary CML and non-hematologic controls were measured by flow cytometry. Cell counting kit-8 analysis, DNA Content Quantitation Assay, and Annexin V-PE/7-AAD staining were applied to assess the effects of CSE1L knockdown on cell proliferation, cell cycle progression, and apoptosis. RESULTS: Elevated expression of CSE1L was detected in bone marrow granulocytes of patients with primary CML. In the CML cell line K562 cells, CSE1L knockdown impaired cell proliferation blocked the cell cycle shift from G0/G1 phase to the S phase, and promoted apoptosis. Knockdown of CSE1L reduced Bcl-2 protein expression and increased Bax protein expression. Meanwhile, knockdown of CSE1L enhanced the expression of phospho-AMPK protein and decreased the expression of phospho-mTOR protein. The expression of total AMPK and mTOR proteins was not affected. In addition, CSE1L expression levels were decreased in imatinib-treated K562 cells. CONCLUSIONS: CSE1L plays a pivotal role in K562 cell survival and growth. These functions may be partially dependent on the AMPK/mTOR signaling pathway to achieve. In addition, CSE1L may have had a future impact on the treatment of CML patients.
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Proteínas Quinasas Activadas por AMP , Leucemia Mielógena Crónica BCR-ABL Positiva , Humanos , Proteínas Quinasas Activadas por AMP/genética , Proteínas Quinasas Activadas por AMP/metabolismo , Proteínas Quinasas Activadas por AMP/farmacología , Transducción de Señal , Serina-Treonina Quinasas TOR/metabolismo , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Apoptosis , Proliferación Celular , Células K562RESUMEN
OBJECTIVE: To explore the expression of cellular apoptosis susceptibility protein ï¼CASï¼ in acute myeloid leukemia ï¼AMLï¼ and its correlation with clinical characteristics. METHODS: The expression of CAS in bone marrow tissue of 54 patients with AML and 24 patients with non-hematological malignant diseases was detected by Western blot and immune-histochemical method, and compared between AML group and control group. Also the relationship of CAS expression in AML and sex, age, WBC count, Hb, platelet count, bone marrow blast cell ratio, ki-67 index, cytogenetic and molecular biological prognostic risk stratification, extramedullary infiltration and other clinical characteristics was analyzed. RESULTS: Western blot showed that the expression of CAS protein in bone marrow biopsies of AML patients was significantly higher than that in control group (P<0.05). Immune-histochemical method revealed that CAS was mainly located in the cytoplasm in both AML group and control group. Among 54 AML patients, 14 patients (25.9%) showed high expression of CAS, while all the 24 patients in the control group showed low expression of CAS. The high expression rate of CAS in AML patients was significantly higher than that in the control group (P<0.05). There were statistically significant differences in prognostic risk stratification and the remission rate of the first chemotherapy between CAS high expression group and CAS low expression group in AML (P<0.05). The proportion of high risk patients and unremission patients after the first chemotherapy in CAS high expression group were significantly higher than those in CAS low expression group (57.1% vs 27.5%, 30.8% vs 7.9%), while the proportion of low risk patients and complete remission patients after the first chemotherapy were significantly lower than those in CAS low expression group (14.3% vs 37.5%, 53.8% vs 84.2%). In AML patients, the ki-67 index of bone marrow tissue in CAS high expression group was higher than that in CAS low expression group (60% vs 50%) (P<0.05). CONCLUSION: CAS is localized in cytoplasm in both AML and non-hematological malignant diseases, and its expression increases in AML. CAS is related to the risk stratification of cytogenetics and molecular biology, the remission rate after the first chemotherapy and ki-67 index in AML, which suggests that CAS may be involved in the occurrence and development of AML.
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Proteína de Susceptibilidad a Apoptosis Celular , Leucemia Mieloide Aguda , Médula Ósea/metabolismo , Proteína de Susceptibilidad a Apoptosis Celular/metabolismo , Humanos , Antígeno Ki-67/metabolismo , Leucemia Mieloide Aguda/tratamiento farmacológico , Pronóstico , Inducción de RemisiónRESUMEN
BACKGROUND: Coronary heart disease (CHD) is a global public health concern, and CHD risk assessment remains a major challenge. Therefore, in this study, the ability of small dense low-density lipoprotein (sdLDL) alone to sufficiently predict CHD risk in the Chinese population was evaluated. METHODS: Patients with CHD (139) and healthy controls (58) were included in this study. Serum sdLDL was measured using the peroxidase method. Other lipid parameters were also determined. RESULTS: The sdLDL level in the CHD group was significantly higher than that in the control group (p < 0.001). Logistic regression analysis revealed that sdLDL was an independent risk factor for CHD. Based on the receiver operating characteristic curves, the area under the curve (AUC) of sdLDL alone for CHD was 0.722. The AUC of triglycerides (TG), high-density lipoprotein (HDL), and sdLDL combined was 0.763, which was larger than that of the independent ones or combinations of any two; however, the value was not significant. CONCLUSIONS: sdLDL alone can predict CHD risk efficiently similar to the combination of TG, HDL, and sdLDL. This finding suggests that sdLDL can be considered as an ideal parameter for the preliminary diagnosis of CHD in Chinese people.
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Enfermedad Coronaria , China/epidemiología , LDL-Colesterol , Enfermedad Coronaria/diagnóstico , Enfermedad Coronaria/epidemiología , Humanos , Factores de Riesgo , TriglicéridosRESUMEN
Testicular germ cell tumors are the most frequent malignancies found in men between 15 and 44 years old. Although cellular apoptosis susceptibility (CAS) was demonstrated to be upregulated in breast cancer and colon cancer, the expression of CAS in the human testis and testicular germ cell tumors remained elusive. In the present study, CAS-positive signals were detected in the normal testicular tissues, cancer adjacent normal testicular tissues, seminoma, yolk sac tumor, and teratoma. Interestingly, the expression level of CAS in testicular germ cell tumors (TGCTs) (but not seminoma) was significantly lower than that of human testicular tissues and cancer adjacent normal testicular tissues, suggesting that decreased CAS contributed to the progression of TGCTs. Notably, the expression of CAS in seminoma was significantly higher than that of in the non-seminomas, consistent with the results from TCGA database. Furthermore, the localization of CAS is mainly restricted in the nucleus in the lesions of normal human testicular tissue and cancer adjacent normal testicular tissue. Although the expression of CAS was not significantly different between normal testicular tissue and seminoma, CAS was more enriched in cytoplasm in seminoma compared to the normal, cancer adjacent tissue and other types of TGCTs. The current results demonstrated reduced expression of CAS in the human testicular germ cell tumors and the CAS translocation from the nuclear to cytoplasm in seminoma, thereby supporting a possible role in normal testis function and in the development of seminoma.
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Proteína de Susceptibilidad a Apoptosis Celular/biosíntesis , Neoplasias de Células Germinales y Embrionarias/metabolismo , Neoplasias Testiculares/metabolismo , Testículo/metabolismo , Carcinoma Embrionario/metabolismo , Citoplasma/metabolismo , Tumor del Seno Endodérmico/metabolismo , Humanos , Inmunohistoquímica , Masculino , Seminoma/metabolismo , Teratoma/metabolismo , Análisis de Matrices TisularesRESUMEN
OBJECTIVE: To clone human PD-1 gene, construct a prokaryotic expression plasmid and express in E. coli. METHODS: The human PD-1 cDNA was cloned by RT-PCR from the total RNA, which was extracted from peripheral blood lymphocyte cell of the patient with chronic hepatitis B. Recombinant PD-1 protein was been expressed and purified after the prokaryotic expression plasmid had been constructed. It was identified by SDS-PAGE, DNA sequencing and amino acid sequencing. RESULTS: The PD-1 gene was cloned and confirmed by DNA sequencing. The recombinant protein was expressed in E. coli. The purified protein was obtained, then been confirmed by amino acid sequencing. CONCLUSION: The human PD-1 gene was successfully cloned and expressed in E. coli, which lays the foundation for further study on the function and application of PD-1.
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Antígenos CD/biosíntesis , Antígenos CD/genética , Proteínas Reguladoras de la Apoptosis/biosíntesis , Proteínas Reguladoras de la Apoptosis/genética , Escherichia coli/genética , Células Procariotas/metabolismo , Secuencia de Aminoácidos , Antígenos CD/química , Antígenos CD/aislamiento & purificación , Proteínas Reguladoras de la Apoptosis/química , Proteínas Reguladoras de la Apoptosis/aislamiento & purificación , Clonación Molecular , Electroforesis en Gel de Poliacrilamida , Vectores Genéticos/genética , Vectores Genéticos/metabolismo , Humanos , Reacción en Cadena de la Polimerasa , Receptor de Muerte Celular Programada 1 , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
A system for the expression of recombinant lumbrokinase (rPI239) was developed in the yeast Pichia pastoris. A total supernatant protein content of 0.174 g/L of high density fermentation broth was obtained. The rPI239 exhibited in vitro fibrinolytic activity. The in vivo activity of rPI239 was measured by prothrombin time, kaolin part thrombin time, thrombin time, and fibrinolytic activity. This work presents the high-density fermentation of rPI239 from P. pastoris and shows that the recombinant protein has similar fibrinolytic activity both in vivo and in vitro.