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1.
Carcinogenesis ; 39(8): 1006-1015, 2018 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-29850766

RESUMO

Pancreatic cancer (PC) is a highly invasive tumor with early metastasis and poor prognosis, yet the mechanisms for tumor progression have not been fully elucidated. Emerging evidence indicates that microRNA-331-3p (miR-331-3p) plays an important role in the progression of diverse human cancers. Here, we found that miR-331-3p was significantly upregulated in tumor specimens of PC patients and PC cell lines. Functional studies showed that downregulation of miR-331-3p inhibited PC cell proliferation and epithelial-mesenchymal transition (EMT)-mediated metastasis in vitro. Furthermore, suppression of tumorigenicity 7 like (ST7L) was identified as a novel target gene of miR-331-3p. Tumor promotion effects of miR-331-3p were partially reversed by ST7L re-expression. In addition, miR-331-3p antagomir suppressed PC tumor growth and metastasis via upregulation of ST7L in xenograft mice. In summary, these results demonstrate that miR-331-3p is a tumor-promoting microRNA (miRNA) in PC cells and a promising biomarker for PC.


Assuntos
Biomarcadores Tumorais/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/metabolismo , Neoplasias Pancreáticas/genética , Proteínas de Ligação a RNA/genética , Idoso , Animais , Biomarcadores Tumorais/antagonistas & inibidores , Linhagem Celular Tumoral , Proliferação de Células/genética , Transição Epitelial-Mesenquimal/genética , Feminino , Humanos , Masculino , Camundongos , MicroRNAs/antagonistas & inibidores , Pessoa de Meia-Idade , Oncogenes , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/patologia , Proteínas de Ligação a RNA/metabolismo , Proteínas Supressoras de Tumor , Ensaios Antitumorais Modelo de Xenoenxerto
2.
Zhonghua Zhong Liu Za Zhi ; 38(1): 28-34, 2016 Jan.
Artigo em Zh | MEDLINE | ID: mdl-26796803

RESUMO

OBJECTIVE: To observe the efficacy and safety of chemotherapy regimens oxaliplatin combined with capecitabine (CAPOX) or oxaliplatin combined with tegafur, gimeracil and oteracil potassium capsules (S-1)(SOX), and to investigate the value of expression of thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) proteins in tumor tissue for predicting the efficacy of CAPOX and SOX regimens in advanced gastric cancer patients. METHODS: A total of 107 newly-diagnosed, stage Ⅲc/Ⅳ gastric cancer patients (no surgical indication, ECOG performance scores 0-2 and expected survival time ≥3 months) were recruited with 101 patients evaluated. The patients were randomly divided into two groups. One was study group in which the patients received CAPOX regimen. The other was control group received SOX regimen. Each patient received four cycles, at least two cycles chemotherapy every three weeks and followed up until death or lost. Tumor biopsies were obtained by gastroscopy for immunohistochemical examination of the expression of TP and DPD proteins before chemotherapy. Response rate (ORR), overall survival (OS) and time to tumor progression (TTP) of the patients were assessed. RESULTS: The objective response rate (ORR) of the study and control groups was 49.0% (5/51) vs. 46.0% (23/50), respectively (P>0.05). The overall survival (OS) was 357.36±24.69 days in the study group and 349.87±22.63 days in the control group, and the time-to-progression (TTP) was 216.75±19.32 days in the study group and 220.54±18.47 days in the control group (P>0.05 for both). Stratified analysis showed that the ORR of TP-positive patients in the study group was significantly higher than that in the control group (72.0 % vs. 41.7 %, P=0.032). There was no significant difference in ORR between the TP-negative patients in the study and control groups (26.9% vs. 50.0%, P=0.087), while the ORR of DPD-positive patients in the control group was significantly higher than that of the study group (51.9% vs. 34.6%, P=0.046). There was no significant difference in the ORR between DPD-negative patients in the study and control groups (64.0% vs. 39.1%, P=0.084). The follow-up showed that the OS (378.42±22.56 days) and TTP (271.77±24.92 days) in the TP-positive patients of the study group were significantly longer than those of the control group (OS: 326.57±19.84 days, and TTP: 229.13±22.68 days)( P<0.05). The OS was 371.25±23.97 days and TTP was 264.66±21.36 days in the DPD-positive patients of control group, significantly longer than those of the study group (OS: 334.73±21.47days, and TTP: 208.58±20.70 days) (P<0.05). But there was no significant difference in the OS and TTP between the TP- and DPD-negative patients in the two groups (P>0.05). In respect of adverse events, both the rates of hematological and non-hematological toxicities were low and similar between the two groups (P>0.05), and well-tolerated by the patients. CONCLUSIONS: Both CAPOX and SOX regimens are effective chemotherapeutic protocols in treatment of patients with advanced gastric cancer. The expression levels of TP and DPD in tumor tissue can be used as a predictive factor for the efficacy of capecitabine or tegafur, gimeracil and oteracil potassium capsules combined with oxaliplatin regimens. CAPOX chemotherapy regimen is more suitable for the TP-positive gastric cancer patients, and SOX regimen is more suitable for the DPS-positive gastric cancer patients.


Assuntos
Antineoplásicos/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias Gástricas/tratamento farmacológico , Antineoplásicos/efeitos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Capecitabina/administração & dosagem , Capecitabina/efeitos adversos , Cápsulas , Di-Hidrouracila Desidrogenase (NADP)/metabolismo , Progressão da Doença , Humanos , Proteínas de Neoplasias/metabolismo , Compostos Organoplatínicos/administração & dosagem , Compostos Organoplatínicos/efeitos adversos , Oxaliplatina , Ácido Oxônico/administração & dosagem , Ácido Oxônico/efeitos adversos , Piridinas/administração & dosagem , Piridinas/efeitos adversos , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/mortalidade , Neoplasias Gástricas/patologia , Tegafur/administração & dosagem , Tegafur/efeitos adversos , Timidina Fosforilase/metabolismo
3.
Environ Sci Pollut Res Int ; 30(41): 94401-94413, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37531060

RESUMO

A novel hierarchically nanostructured magnetite (Fe3O4) was manufactured using microwave-assisted reflux method without surfactants. The nanostructured Fe3O4 is formed via the co-precipitation of Fe(III) and Fe(II), followed by a nanocrystal aggregation-based mechanism. Moreover, the effects of solution pH, contact time, initial Sb concentration, coexisting anions, and recycle numbers on the adsorption of nanostructured Fe3O4 toward Sb were extensively examined in the batch adsorption tests. The results demonstrated that the obtained Fe3O4 exhibited excellent adsorption ability toward Sb with the maximum adsorption capacities of 154.2 and 161.1 mg.g-1 for Sb(III) and Sb(V), respectively. The prepared Fe3O4 could be easily regenerated and reused for adsorption/desorption studies multiple times without compromising the Sb adsorption ability. Further exploration indicated that the oxidation or reduction reactions infrequently occurred during Sb adsorption processes. The proposed hierarchically nanostructured Fe3O4 thus could be potentially used for sustainable and efficient antimony removal.


Assuntos
Antimônio , Poluentes Químicos da Água , Antimônio/química , Compostos Férricos/química , Adsorção , Micro-Ondas , Oxirredução
4.
Zhonghua Nei Ke Za Zhi ; 50(6): 469-73, 2011 Jun.
Artigo em Zh | MEDLINE | ID: mdl-21781528

RESUMO

OBJECTIVE: To investigate the predictive value of breast cancer susceptibility gene 1 (BRCA1) and class IIIß-tubulin protein expression in tumor tissue for the efficacy of taxol and cisplatin combined chemotherapy (TP) in stage IIIB/IV non-small cell lung cancer (NSCLC) patients. METHODS: A total of 92 stage IIIB/IV NSCLC patients were recruited with 87 patients evaluated. Bronchoscopy or lung puncture tumor biopsy samples were obtained with BRCA1 and class IIIß-tubulin protein expression examined immunohistochemically before chemotherapy. The patients were randomly assigned to be received 4 to 6 cycles of TP chemotherapy regiments and followed up until death or lost. Response rate (RR), overall survival (OS) and time to tumor progression (TTP) were assessed. RESULTS: Among the 87 evaluated patients, the positive expression rates of BRCA1 and class IIIß-tubulin were 57.5% (50/87) and 48.3% (42/87) respectively. There was no significant difference in clinical characteristics among patients with different positive expression rate. According to different expression of BRCA1 and class IIIß-tubulin, the patients were divided into four groups: group A (low expression of both BRCA1 and class IIIß-tubulin), group B (high expression of both BRCA1 and class IIIß-tubulin), group C (high expression of only BRCA1) and group D (high expression of only class IIIß-tubulin). The RR was higher in group A than other three groups (60.7%, 34.8%, 9/19 and 6/17 respectively). The OS and TTP were longer in group A than other three groups [OS: (539.4 ± 17.6) days, (267.2 ± 20.5) days, (325.6 ± 24.1) days and (283.7 ± 26.2) days respectively; TTP: (256.9 ± 28.4) days, (143.8 ± 17.6) days, (179.3 ± 19.8) days and (152.6 ± 23.5) days respectively]. There were no significant differences among the other three groups. CONCLUSIONS: The expression level of BRCA1 and class IIIß-tubulin in tumor tissue is probably a predictor for the efficacy of TP chemotherapy in NSCLC patients. TP chemotherapy is more suitable for the NSCLC patients with lower expression of both BRCA1 and class IIIß-tubulin. Our study may provide a new sight for tailored chemotherapy in NSCLC patients.


Assuntos
Proteína BRCA1/metabolismo , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Neoplasias Pulmonares/metabolismo , Tubulina (Proteína)/metabolismo , Adulto , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/patologia , Feminino , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento
5.
Ying Yong Sheng Tai Xue Bao ; 32(9): 3257-3266, 2021 Sep.
Artigo em Zh | MEDLINE | ID: mdl-34658212

RESUMO

In order to understand the composition and stability of soil aggregate in paddy filed, as well as the changes of soil aggregate-associated nitrogen (N), phosphorus (P) and potassium (K) after straw addition combined with chemical fertilization, soil samples were collected from a 34-year positioning experiment with three treatments, including no chemical fertilizer (CK), chemical fertilizer only (NPK), and straw addition plus chemical fertilizer (NPKS). The composition of water-stable aggregates at the soil layers of 0-20 cm and 20-40 cm were analyzed with the wet sieving method, as well as the distribution characteristics, contribution rate and activation rate of soil aggregate-associated N, P, and K. Results showed that the fractions of >2 mm and 0.25-1 mm aggregates dominated the soil water-stable aggregates in paddy field, while the contribution of <0.053 mm aggregates was lowest. Compared with CK, NPKS treatment increased the contents of >2 mm and 1-2 mm aggregates at the layers of 0-20 and 20-40 cm, and reduced the contents of 0.053-0.25 mm and <0.053 mm. Similar result in NPK treatment was observed at the layer of 0-20 cm. Compared with tat under the NPK treatment, mean weight diameter (MWD) and geometric mean diameter (GMD) increased by 3.9%-15.5% and 6.3%-41.7% in NPKS treatment, respectively. However, the unstable aggregate index (ELT) reduced by 5.7%-28.7% in the NPKS treatment. NPKS significantly increased the contents of total N (TN), available P (AP), and available K (AK) in soil aggregates, especially in the >0.25 mm aggregates. There were no significant diffe-rences about alkali-hydrolysable N (AN) and total K (TK) between NPK and NPKS treatments. The nutrient contribution of soil aggregates in paddy field was affected by aggregate composition. NPKS significantly increased the contribution of AN, AP, and AK within >1 mm aggregates. In all, straw addition combined with chemical fertilizer could increase the stability of soil aggregates at the layers of 0-20 cm and 20-40 cm, and increase the contents of soil aggregate-associated N, P and K, especially for the >1 mm aggregates. Our results provided insights into ensuring soil quality and sustainable development of resources in paddy field by adjusting the ratio of soil C to N.


Assuntos
Nitrogênio , Fósforo , Agricultura , Fertilizantes , Potássio
6.
Mol Ther Oncolytics ; 18: 432-442, 2020 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-32913892

RESUMO

Pancreatic cancer cells are characterized by high cell proliferation and low cell apoptosis, but the factors involved in these processes remain to be further studied. In this study, we report that miR-324-5p regulates the proliferation and apoptosis of pancreatic cancer cells through regulating the expression of Krüppel-like factor 3 (KLF3). In both pancreatic cancer tissues and cell lines, the levels of miR-324-5p are significantly increased. Inhibition of miR-324-5p represses cell proliferation but promotes cell apoptosis, whereas overexpression of miR-324-5p exerts the opposite effect. Furthermore, we identified KLF3, a factor regulating pancreatic cancer cell proliferation and apoptosis, as a new direct downstream target of miR-324-5p. Our results suggest that miR-324-5p plays an important role in pancreatic cancer cell proliferation and apoptosis via downregulating the expression of KLF3.

7.
Zhonghua Yi Xue Za Zhi ; 89(20): 1382-6, 2009 May 26.
Artigo em Zh | MEDLINE | ID: mdl-19671326

RESUMO

OBJECTIVE: To study the effect of allicin on invasion and metastasis of human colon cancer cell line LoVo in vitro and furthermore elucidate its anticancer mechanisms. METHODS: MTT assay was used to test dynamically the effect of cell growth inhibition. The inhibitory effects of allicin on movement, adhesiveness and invasiveness of LoVo cells were evaluated by the migratory test, adhesion test and Transwell chamber experiment. Quantitative real-time reverse transcription PCR (real-time RT-PCR) was performed to quantify the mRNA expression of MMP-2, TIMP-2, CD147, VEGF, nm23-H1, HPA and uPAR. RESULTS: Allicin had inhibitive effects on growth of LoVo cells in a dose and time-dependent manner. Allicin at non-cytotoxic concentration (3 and 6 microg/ml) could obviously suppress the movement, adhesion and invasive capability of LoVo cells. In the allicin-treated group (3 and 6 microg/ml), after 24 hours, the inhibition rates of migratory time were 24% and 50% (t = 4.543, 12.348, P = 0.010, 0.001), the inhibition rates of adhesion were 19% and 28% (t = 6.145, 6.355, P = 0.004, 0.003), the inhibition rates of migration were 28% and 46% (t = 8.065, 28.435, both P < 0.01), and the inhibition rates of invasion were 44% and 65% respectively (t = 21.274, 26.288, both P < 0.01). Allicin at non-cytotoxic concentration could down-regulate the mRNA levels of VEGF, uPAR and HPA in a dose-dependent manner in LoVo cells (t = 7.129, 6.764, 8.497, P = 0.002, 0.002, 0.001) while the mRNA levels of TIMP-2, CD147 and nm23-H1 remained basically unchanged with the same treatment (t = 0.341, 1.889, 0.914, P = 0.059, 0.132, 0.412). The expression of MMP-2 had not been detected in LoVo cells. CONCLUSION: Allicin in vitro inhibits invasion and metastasis of human colon carcinoma cell LoVo at non-cytotoxic concentration through down-regulating the expression of VEGF, uPAR and HPA mRNA.


Assuntos
Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Ácidos Sulfínicos/farmacologia , Adesão Celular , Linhagem Celular Tumoral , Proliferação de Células , Dissulfetos , Humanos , Metaloproteinase 2 da Matriz/metabolismo , Invasividade Neoplásica , Metástase Neoplásica , RNA Mensageiro/genética , Receptores de Ativador de Plasminogênio Tipo Uroquinase/metabolismo , Inibidor Tecidual de Metaloproteinase-2/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
8.
Pathol Res Pract ; 215(10): 152553, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31362888

RESUMO

BACKGROUND: Nidogen-2 (NID2) is a ubiquitous component in the basement membrane and plays an important role in the development of malignant tumors. However, the specific function and mechanism of the NID2 gene in gastric cancer remains unclear. In this study, we aimed to investigate the role of NID2 in gastric cancer(GC). METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of NID2 in 67 GC tissues and adjacent normal tissues. The relationship between NID2 expression and clinicopathological features was further analyzed. In addition, we evaluated the expression of NID2 in GC based on data from the GEPIA and Kaplan-Meier Plotter database and compared the database results with our own experimental results. Invasion and wound healing assays were used to detect the function of NID2 in MKN45 and SGC7901 cells. Finally, the NID2 network and its possible related genes are constructed by the bioinformatics framework. RESULTS: The expression level of NID2 was found to be significantly over-expressed in gastric cancer cells and tissues compared with normal controls and positively associated with TNM stage, showing a poor prognosis of GC patients. In vitro experiments indicated that NID2 was able to promote the ability of invasion and migration in GC cells. Bioinformatics prediction showed NID2 might regulate the progression of GC via protein digestion and absorption, amoebiasis, PI3K-AKt-signaling pathway, focal adhesion and ECM-receptor interaction pathways. CONCLUSION: Our study demonstrates that up-regulated NID2 plays an important role in promoting the invasion and migration of GC cells and has a potential of being a novel biomarker for diagnosis, treatment and prognosis of GC in the future.


Assuntos
Biomarcadores Tumorais/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Moléculas de Adesão Celular/metabolismo , Movimento Celular/fisiologia , Invasividade Neoplásica/patologia , Neoplasias Gástricas/patologia , Estômago/patologia , Proliferação de Células , Bases de Dados Factuais , Progressão da Doença , Intervalo Livre de Doença , Feminino , Mucosa Gástrica/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/mortalidade , Taxa de Sobrevida , Regulação para Cima , Cicatrização/fisiologia
9.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 25(5): 1550-1558, 2017 Oct.
Artigo em Zh | MEDLINE | ID: mdl-29070142

RESUMO

OBJECTIVE: To investigate the effect of P2X7R antagonist brilliant blue G (BBG) on aGVDH of mice after allo-HSCT. METHODS: aGVHD mouse model after HSCT was established and treated with the P2X7R antagonist BBG of different dosages (50 mg/kg and 75 mg/kg). After treatment, the survival, body weight, pathological and liver function of aGVDH mice were abserved, and the expression levels of P2X7, NLRP3, caspase-1, IL-1ß, IL-18 mRNA and protein were evaluated by real-time PCR and Western blot. RESULTS: The allo-HSCT aGVHD mouse model was successfully established, the intraperitoneal injection of BBG alleviated the aGVHD clinical manifestations including roachback, ruffled fur, skin peeling and weight loss of recipient mice, decreased P2X7R and IL-1ß expression and reduced the mRNA levels of P2X7R, NLRP3, Caspase-1, IL-1ß and IL-18. Furthermore, GVHD group receiving 75 mg/kg BBG showed most significant difference of these indexes. CONCLUSION: BBG alleviates liver inflammatory damage induced by aGVHD after allo-HSCT, and decreases the expression of proinflammatory cytokines. Moreover, the protective effect of that of BBG 75 mg/kg group is better than that of BBG 50 mg/kg group.


Assuntos
Doença Enxerto-Hospedeiro , Transplante de Células-Tronco Hematopoéticas , Corantes de Rosanilina/uso terapêutico , Doença Aguda , Animais , Citocinas , Modelos Animais de Doenças , Doença Enxerto-Hospedeiro/tratamento farmacológico , Doença Enxerto-Hospedeiro/metabolismo , Interleucina-1beta , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR
10.
Oncol Rep ; 33(3): 1402-10, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25607234

RESUMO

Colorectal cancer (CRC) is the most common cancer diagnosed worldwide, and the development of metastases is a major cause of mortality. Accumulating evidence suggests that microRNAs are important in carcinogenesis by affecting the expression of genes that regulate cancer progression. A number of studies have shown that miR-206 is frequently downregulated in many human malignancies, including CRC, and is associated with a more malignant phenotype. Previous studies involving HeLa and C2C12 cells have validated the inhibitory mechanism of miR-206 via NOTCH3 targeting. However, whether or not the interplay between miR-206 and NOTCH3 also occurs in CRC is unknown. Therefore, we investigated the tumor suppressive and metastatic effects of miR-206 and its target, NOTCH3, in CRC. Based on the inverse association between the expression of miR-206 and NOTCH3 in CRC tissues, miR-206 mimics were transiently transfected into the SW480 (and its metastatic strain) and SW620 colon cancer cell lines. Upregulation of miR-206 inhibited cancer cell prolife-ration and migration, blocked the cell cycle, and activated apoptosis. The tumor suppressive capacity of miR-206 had a similar effect on CRC cells, although with a different metastatic potential, and may be explained by direct NOTCH3 signaling inhibition and indirect cross-talk with other signaling pathways involving CDH2 and MMP-9. These results support miR-206 as a tumor suppressor in CRC and suggest a potential therapeutic target for clinical intervention.


Assuntos
Apoptose/genética , Neoplasias Colorretais/genética , MicroRNAs/genética , Receptores Notch/antagonistas & inibidores , Antígenos CD/metabolismo , Caderinas/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Neoplasias Colorretais/patologia , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica , Genes Supressores de Tumor , Humanos , Metaloproteinase 9 da Matriz/metabolismo , MicroRNAs/biossíntese , Receptor Notch3 , Receptores Notch/biossíntese , Transfecção , Cicatrização/genética
11.
Ai Zheng ; 25(6): 718-22, 2006 Jun.
Artigo em Zh | MEDLINE | ID: mdl-16764767

RESUMO

BACKGROUND & OBJECTIVE: Vascular endothelial growth factor (VEGF) signal pathway is a hot spot in recent tumor research, but its role in rhabdomyosarcoma (RMS) has rarely been reported, and its mechanism is unclear. This study was to investigate the expression of VEGF and its receptors (VEGFR) in RMS cell line RH4 to reveal the mechanism of VEGF signal pathway, and explore the therapeutic value of Avastin in RMS in vitro. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of VEGF, VEGFR1, VEGFR2, and VEGFR3 mRNA in RH4 cells. ELISA was used to detect the concentration of VEGF in supernatant. Western blot was used to detect the expression of VEGFR1 protein. The effects of VEGF and Avastin on RH4 cell growth were evaluated by direct cell counting. RESULTS: RH4 cells expressed VEGF at a high level and secreted VEGF to culture media. RH4 cells only expressed VEGFR1, did not express VEGFR2 and VEGFR3. VEGF promoted the growth of RH4 cells in a dose-dependent manner within a concentration range of 0-100 ng/ml, this effect could be blocked by 10-40 microg/ml Avastin. CONCLUSION: VEGF can promote the growth of RMS cell through VEGFR1, and this effect can be blocked by Avastin.


Assuntos
Anticorpos Monoclonais/farmacologia , Proliferação de Células/efeitos dos fármacos , Rabdomiossarcoma/patologia , Fator A de Crescimento do Endotélio Vascular/farmacologia , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais Humanizados , Bevacizumab , Western Blotting , Técnicas de Cultura de Células , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Humanos , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rabdomiossarcoma/metabolismo , Fator A de Crescimento do Endotélio Vascular/administração & dosagem , Fator A de Crescimento do Endotélio Vascular/biossíntese , Fator A de Crescimento do Endotélio Vascular/genética
12.
Ai Zheng ; 24(11): 1408-11, 2005 Nov.
Artigo em Zh | MEDLINE | ID: mdl-16552973

RESUMO

BACKGROUND & OBJECTIVE: Recently, chitosan, as a nonviral vehicle for transferring DNA molecules into the cells, has attracted much attention because of its cationic properties. This study was to investigate characteristics and transfection activity of chitosan-pDNA microparticles, and confirm the feasibility of chitosan as gene therapy vehicle, which may be employed in further in vivo study. METHODS: Plasmid DNAs were amplified in Eschericha Coli JM109 with transfection of enhanced green fluorescence protein (EGFP) gene, and isolated according to the protocol of Qiagen plasmid midi kit. Chitosan-pDNA microparticles were prepared by the coacervation method, and observed under transmission electronic microscope. In vitro releasing experiment and ultraviolet spectrophotometry were used to measure DNA loading capacity and loading efficiency. The location of pDNA in the microparticles was analyzed by gel retardation assay. Transfection efficiency of chitosan-pDNA microparticles was evaluated by detecting gene expression of EGFP in HEK293 cells after transfection. RESULTS: Chitosan-pDNA microparticles were global, and the size ranged 100-200 nm, with the average of (138 +/- 43) nm. DNA loading capacity and loading efficiency of the microparticles were (46.8 +/- 9.0)% and 100%, respectively. Gel retardation assay confirmed that DNA was wholly encapsulated in the microparticles. In vitro study revealed that the release of DNA from chitosan-pDNA microparticles included two-step process, namely burst effect with a slow second phase. Most of DNA could be released at the time of 48 h. In vitro transfection results indicated that chitosan could efficiently deliver pEGFP into HEK293 cells and stably express green florescence protein. CONCLUSIONS: Chitosan can efficiently transfer target DNA molecules into mammalian cells, continuously release DNA, and stably express DNA. It can be potentially employed as a gene therapy vehicle.


Assuntos
Quitosana/administração & dosagem , Sistemas de Liberação de Medicamentos , Linhagem Celular , Quitosana/química , DNA/metabolismo , Portadores de Fármacos , Estudos de Viabilidade , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Rim/citologia , Rim/metabolismo , Microesferas , Tamanho da Partícula , Plasmídeos , Transfecção
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