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1.
Mol Cancer Ther ; 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31582530

RESUMO

Viral-based CAR T cell manufacturing has potential safety risks and relatively high costs. The non-viral minicircle DNA (mcDNA) is safer for patients, cheaper to produce, and may be a more suitable technique to generate CAR T cells. In this study, we produced mcDNA-based CAR T cells specifically targeting PSCA (mcDNA-PSCA-CAR T cells). Our results showed that mcDNA-PSCA-CAR T cells persisted in mouse peripheral blood as long as 28 days and demonstrated more CAR T cell infiltration, higher cytokine secretion levels, and better anti-tumor effects. Together, our results suggest that mcDNA-CAR can be a safe and cost-effective platform to produce CAR T cells.

2.
SLAS Discov ; : 2472555219878407, 2019 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-31560248

RESUMO

D-dimer is an essential diagnostic index of thrombotic diseases. Since the existing anti-D-dimer antibodies vary in quality and specificity, a search for alternative anti-D-dimer antibodies is required. The present study aimed to screen a novel monoclonal antibody (mAb) against D-dimer using a light-initiated chemiluminescence assay (LiCA). In this work, mice were immunized with antigen prepared from human plasma by enzyme hydrolysis. After screening, a novel mAb, DD 2G11, was obtained. The results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis indicated that DD 2G11 could be used as a standard marker for D-dimer. The isotype of DD 2G11 was IgG1, the Ka value was 0.646 nM-1, and the Kd value was 50 nM, indicating that the binding affinity to D-dimer was very high. Furthermore, no cross-reactivity between DD 2G11 and other fibrinogen degradation products (FgDPs) was found. Finally, the correlation between DD 2G11 and the reference antibody (commercial antibody) was investigated by analyzing 56 clinical samples using a latex-enhanced turbidimetric immunoassay (LTIA). The R2 value of the linear regression was 0.94538, indicating that DD 2G11 met clinical requirements. In conclusion, the present study provides a more expeditious protocol to screen mAbs and provides a clinically usable mAb against D-dimer.

3.
FEBS Lett ; 593(18): 2566-2573, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31254364

RESUMO

Aquaporin 1 (AQP1) plays an important role in endothelial functions and is regulated by MEF2C. However, how AQP1 level is stabilized to maintain endothelial water homeostasis is still not clear. Here, we show that AQP1 expression is significantly upregulated by MEF2C transcriptionally and inhibited by miR-133a-3p.1 post-transcriptionally. Meanwhile, MEF2C activates the expression of miR-133a1. Simultaneous overexpression of MEF2C and miR-133a-3p.1 suppresses the aptitude of changes in AQP1 expression caused by either MEF2C or miR-133a-3p.1. Accordingly, the changes in migration and tube formation of human umbilical vein endothelial cells (HUVECs) caused by MEF2C or miR133a-3p.1 are blunted by coexpression of both of them. These data demonstrate that the homeostasis and physiological function of AQP1 in endothelial health are maintained by the MEF2C and miR-133a-3p.1 regulatory circuit.

4.
FEBS Open Bio ; 9(7): 1305-1314, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31141316

RESUMO

Aberrant expression of microRNAs (miRNAs) may contribute to the initiation and development of multiple types of human cancer. Several miRNAs have been found to be strongly correlated with the diagnosis, progression, and prognosis of colorectal carcinoma (CRC), but the role of miR-125a in CRC remains unclear. In the present study, the function of miR-125a on the expression of Smad ubiquitin regulatory factor 1 (Smurf1) was investigated in vitro and in vivo. We verified that Smurf1 is a downstream target gene of miR-125a and is involved in miR-125a-mediated regulation of CT26 cell (colon cancer cell) proliferation and migration. Overexpression of miR-125a suppresses CT26 cell growth by inhibiting cell proliferation. Additionally, wound healing assays were performed to show that overexpression of miR-125a significantly reduced CT26 cell migration, which was reversed by overexpression of Smurf1. In vivo, miR-125a overexpression downregulated the expression of Ki67 and Smurf1, thus leading to a marked reduction in tumor growth. These results revealed that miR-125a plays a critical role in CRC by directly targeting Smurf1, a finding that may facilitate the development of improved diagnostic and therapeutic techniques for CRC.

5.
Immunotherapy ; 11(7): 599-616, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30943862

RESUMO

AIM: To produce dendritic cells (DCs) from CD34+ stem cells from cord blood and explore their prophylactic and curative effect against tumors by vaccinating humanized NSG mice. MATERIALS & METHODS: Separated CD34+ stem cells from cord blood were cultured for 30 days, and the resultant DCs (CD34-DCs) were collected. The basic function of the CD34-DCs and the cytotoxicity of CD34-cytotoxic-T lymphocytes (CTLs) were tested in vitro, and tumor inhibition in a humanized NSG mouse tumor model was observed. RESULTS: The number of CD34-DCs reached approximately 9 log. These cells performed functions similar to those of DCs derived from monocytes from peripheral blood (PBMC-DCs). The CTLs of the CD34-DCs (CD34-CTLs) presented a better antitumor effect in vitro. The obvious prophylactic and therapeutic antitumor effects of the CD34-DC vaccine were observed in the humanized NSG mouse models. CONCLUSION: CD34-DCs from cord blood were sufficient in quantity and quality as a vaccine agent against tumors in vitro and in vivo.

6.
Biomed Pharmacother ; 114: 108708, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30913493

RESUMO

PURPOSE: Dynamic remodeling of the extracellular matrix (ECM) around tumor cells is crucial for the tumor progressions. However, the mechanism is not well defined. Here, we aimed to reveal the underlying mechanism of ECM induced metastasis and provide innovative strategy to suppress the distant metastasis induced by ECM. MATERIALS AND METHODS: IHC was used to detect the expression of target proteins. H&E staining was used to evaluate the growth of tumor in vivo. Using wound healing and transwell assay, we examined the ability of cell to metastasis. We employed IF and Western blot to detect the expression of target proteins. And qRT-PCR was used to examine the target genes in mRNA level. We also applied flow cytometry to examine the percent of CD133+ cell population. RESULTS: Herein, we observed elevated expression of type I collagen in colorectal cancer tissues from patients with high metastasis. Additionally, colorectal cancer cells cultured on 2D collagen reveal obviously enhanced capability of metastasis and tumorigenesis both in vitro and in vivo. We demonstrated that the activation of PI3K/AKT signal induced by integrin α2ß1 resulted in the enhanced metastatic capability and stemness of colorectal cancer cells. Moreover, we found that Snail worked as the downstream of PI3K/AKT signaling, resulting in the intensive invasion and metastasis of colorectal cancer. Blocking the pathway by applying E7820 successfully reversed the type I collagen induced distant metastasis in colorectal cancer. CONCLUSION: Combining E7820 and chemotherapeutic agents to block the integrin α2ß1/PI3K/AKT/Snail signaling pathway revealed dramatic enhanced tumor suppression and provided an innovative approach for clinical colorectal cancer treatment.


Assuntos
Colágeno/metabolismo , Neoplasias Colorretais/metabolismo , Integrinas/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/fisiologia , Fatores de Transcrição da Família Snail/metabolismo , Animais , Carcinogênese/metabolismo , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Transição Epitelial-Mesenquimal/fisiologia , Feminino , Células HCT116 , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus
7.
Cancer Chemother Pharmacol ; 83(5): 911-920, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30848330

RESUMO

Activation of programmed death-1 (PD-1) and cytotoxic T-lymphocyte antigen-4 (CTLA-4) on T cells leads to T cell exhaustion and ultimately facilitates tumor progression. Recent success of using immune cell checkpoint inhibitors offers a great promise to treat various cancers, including bladder cancer. However, the expression pattern and therapeutic value of PD-1 and CTLA-4 in peripheral blood T cells remain largely unexplored. In this study, we presume that disruption of the potential dysregulated checkpoint molecules in peripheral blood T cells may improve the anti-tumor efficacy of cytotoxic T cells in bladder cancer. We showed that both PD-1 and CTLA-4 expression were specifically elevated on CD8 + T cells but not CD4 + T cells in peripheral blood of patients with bladder cancer compared with that in healthy donors. Notably, CTLA-4 expression was significantly higher in muscle-invasive bladder cancer (MIBC) and correlated with tumor size. By blocking CTLA-4 with anti-CTLA-4 antibody and CRISPR-Cas9-mediated CTLA-4 disruption, we revealed that CTLA-4-disrupted CTLs had enhanced cellular immune response and superior cytotoxicity to the CD80/CD86-positive bladder cancer cells in vitro. Moreover, the CTLA-4-disrupted CTLs exhibited a pronounced anti-tumor effect in vivo as demonstrated by prophylactic assay and therapeutic assay in the subcutaneous xenograft model. Collectively, our findings confirm improved therapeutic efficacy of CTLA-4-disrupted CTLs and provides the potential strategy for targeting immune checkpoints to enhance the promising immunotherapy.

8.
Eur J Pharm Sci ; 130: 78-90, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30684657

RESUMO

Many strategies have been employed to improve oral drug delivery. One such approach involves the use of supersaturable delivery systems such as amorphous self-micellizing solid dispersions (SmSDs). SmSDs have attracted more attention recently, but little is known regarding the impact of production methods on profiles and internal mechanisms of final SmSDs in spite of its importance. In this study, amorphous SmSDs containing self-micellizing Soluplus® and BCS II drug (either indomethacin (IND) or fenofibrate (FEN)) were generated using various methods: solvent evaporation (SOL), freeze-drying (FD), microwave radiation-quench cooling (MQC), and hot melt extrusion (HME). Microscopic morphology, amorphous state, thermal behavior, dissolution/solubility, and "spring-parachute" data were used to assemble physicochemical profiles for SmSD systems prepared using each method. Analysis of intermolecular interactions, solubilization, and crystallization inhibition further uncovered internal mechanisms explaining observed physicochemical properties. Generally, SmSD/IND and SmSD/FEN systems generated using HME exhibited superior dissolution, solubility, and spring-parachute profiles. The superior advantages of HME-generated SmSD/IND systems were attributed to relatively stronger intermolecular interactions than observed in SmSD/IND systems fabricated using other methods. Moreover, self-micellizing Soluplus® carrier was able to solubilize IND or FEN and suppress drug crystallization from a supersaturated state, which seemed to be an important mechanism for the properties enhancement caused by SmSD/FENHME. This knowledge should be useful for guiding further development of self-micellizing solid dispersions and for gaining deeper understanding of how HME technology can improve supersaturable drug delivery based on SmSDs strategy.


Assuntos
Química Farmacêutica/métodos , Fenofibrato/química , Temperatura Alta , Indometacina/química , Micelas , Polietilenoglicóis/química , Polivinil/química , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/farmacocinética , Relação Dose-Resposta a Droga , Fenofibrato/farmacocinética , Hipolipemiantes/química , Hipolipemiantes/farmacocinética , Indometacina/farmacocinética , Polietilenoglicóis/farmacocinética , Polivinil/farmacocinética , Solubilidade , Espectroscopia de Infravermelho com Transformada de Fourier/métodos
9.
Front Pediatr ; 6: 293, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30356669

RESUMO

Purpose: To assess the outcomes of a novel laparoscopic assisted transcrotal orchidopexy (LATO) combined with percutaneous extraperitoneal closure (PEC) for palpable inguinal canalicular cryptorchidism accompany with indirect inguinal hernia, and evaluate its safety and efficiency. Materials and Methods: A retrospective cohort study for single-port LATO-PEC and traditional inguinal orchidopexy (TIO) was performed between 2011 and 2014. Totally 53 children with both palpable inguinal canalicular testes and indirect inguinal hernia were included. Median patient age was 15month (range, 6 months to 4 years). Of them, 35 patients underwent LATO-PEC procedure, utilizing an umbilical trocar for laparoscope, transcrotal dissection for orchidopexy, and an inner two-hooked cannula for ligation of the patent processus at the level of the internal ring. Three of them were bilateral, 12 on the left side and 20 on the right. Eighteen patients received TIO, seven of them on the left side and 11 on the right. Patient demographics, surgical technique, complications, and clinical outcomes were reviewed. Follow-up visits were performed to reassess position and size of the testes. Results: All 56 undescended testes were delivered into the scrotum successfully. In the LATO-PEC group, nine contralateral herniorrhaphy were accomplished simultaneously. Fifteen contralateral patent processus vaginalis (PPVs) in 32 unilateral undescended testis (UDT) were newly confirmed during the laparoscopy, while 6 of them received percutaneous extra-peritoneal herniorrhaphy for visible inguinal bubble in pneumoperitoneum condition. No additional port placement or conversion to open procedure was needed. Mean operative time for unilateral and bilateral LATO-PEC in this study was (37.81 ± 5.23) min and (53.33 ± 2.98) min, respectively. In TIO group, mean operative time was (41.11 ± 8.67) min. There was no statistical difference in operative time between the two approaches for unilateral UDTs (p = 0.098). Median follow-up interval was 24 months (range, 12-84 months). No operative complications were found in either group to date. Conclusions: Singe-port LATO-PEC is a safe, effective, and cosmetic choice for inguinal canalicular cryptorchidism accompany with indirect inguinal hernia, minimizing injuries to the vas deferens and testicular vessels. Laparoscopy can provide a diagnostic and therapeutic solution of contralateral PPV.

10.
Cancer Res Treat ; 2018 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-29690747

RESUMO

Purpose: We investigated the role of tumor-associated macrophages (TAMs) on the epithelial to mesenchymal transition (EMT) of colorectal cancer cells and determined the potential mechanism involved in the metastatic process. Materials and Methods: In this study, flow cytometry was used to detect the expression of target proteins. We used transwell assay to evaluate the migration of cancer cells under specific conditions. Using real-time polymerase chain reaction, we examined the expressions of cytokines and EMT-related markers in mRNA level. Animal assay was performed for analysis in vivo and hematoxylin and eosin was used to visualize the effect of TAMs on tumor metastasis. We also used immunohistochemistry and Western blotting to detect the expression of target proteins. Results: Here, we observed enrichment of TAMs in colorectal tumor tissues, resulting in high metastasis in clinical therapy. Moreover, those TAMs could facilitate the EMT progression of colorectal cancer cells, which is induced by the transforming growth factor-ß (TGF-ß) derived from TAMs, leading to the invasion and migration of cancer cells. Conclusion: Our results demonstrated that TAMs contributed the EMT progression through a TGF-ß/Smad2,3-4/Snail signaling pathway, and disrupting this pathway with TGF-ß receptor inhibitor could suppress metastasis, readjusting our focus to the connection of TAMs and cancer metastasis.

11.
Sci Rep ; 8(1): 2561, 2018 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-29416099

RESUMO

A murine monoclonal antibody (MAb-1) specific for GM3 has been generated by immunizing ß3Gn-T5 knockout mice with purified GM3 ganglioside. The binding specificity of MAb-1 (IgG3 subclass) was established by an enzyme-linked immunosorbent assay (ELISA) and FACS and the antibody showed high binding specificity with GM3. Cell viability assay showed that MAb-1 significantly suppressed cell growth. Immunohistochemistry analysis revealed that MAb-1 was strongly expressed in human ovarian cancer tissues, whereas it was hardly expressed in normal tissues. Finally, antibody-dependent cellular cytotoxicity (ADCC) activities were determined by measuring lactate dehydrogenase (LDH) releasing assay and the results showed high ADCC activities in two representative ovarian cancer cell lines (OVHM and ID8). All of these data indicate that MAb-1 may be potentially used as a therapeutic antibody against ovarian cancers in clinical trials.

12.
Exp Ther Med ; 15(2): 1819-1824, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29434770

RESUMO

Ovarian cancer is a highly prevalent cancer among women. Recent studies have indicated that microRNAs (miRs) may serve important roles in the pathogenesis of ovarian cancer. miR-519a was observed to be downregulated in tissue samples of patients with ovarian cancer; however, its role in ovarian cancer requires further investigation. The aim of the present study was to examine the role of miR-519a in the pathogenesis of ovarian cancer and determine its direct target. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to examine the expression of miR-519a in 20 patients ovarian cancer and 20 normal ovarian tissue samples. Subsequently, SKOV3 cells were cultured and transfected with miR-519a mimics, while MTT and Annexin V assays were performed to investigate the role of miR-519a in the proliferation and apoptosis of SKOV3 cells. In addition, RT-qPCR and western blotting were used to determine the expression levels of miR-519a, signal transducer and activator of transcription 3 (STAT3), myeloid cell leukemia 1 (Mcl-1) and B-cell lymphoma-extra large (Bcl-xl) in untransfected and miR-519a mimic-transfected SKOV3 cells. Dual-luciferase reporter assay was also performed to confirm whether STAT3 was a direct target of miR-519a. The results revealed that miR-519a was significantly downregulated in tissue samples of patients with ovarian cancer as compared with the normal ovarian tissues. Furthermore, transient overexpression of miR-519a inhibited the proliferation and promoted the apoptosis of SKOV3 cells, as well as decreased the mRNA and protein expression levels of STAT3, Mcl-1 and Bcl-xl. Finally, dual-luciferase reporter assay confirmed that STAT3 was a direct target of miR-519a. In conclusion, the present study proved for the first time that miR-519a functions as a tumor suppressor by targeting STAT3 in ovarian cancer, suggesting that miR-519a may be a potential biomarker for the diagnosis and treatment of ovarian cancer.

13.
Oncotarget ; 9(4): 5208-5215, 2018 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-29435173

RESUMO

Programmed cell death protein 1 (PD-1) is an immune checkpoint receptor that functions to attenuate T cell activation. In this study, we knocked out (KO) PD-1 in cytotoxic T lymphocytes (CTLs) using CRISPR-Cas9 system to evaluate its effect on the anti-tumor activity of the CTLs against multiple myeloma (MM). Results show that PD-1 KO CTLs facilitate apoptosis and caspase activation of the co-cultured MM cells and enhanced MM cell death by 36% compared with the control. PD-1 KO also increased TNF-α and IFN-γ secretion of the CTLs by 2.4 and 1.9-fold respectively. The effectiveness of PD-1 KO in enhancing anti-tumor activity of the CTLs was verified in vivo using mouse xenograft model. The xenografted mice treated with PD-1 KO CTLs demonstrated repressed MM tumor growth and prolonged survival compared with the control group. We conclude that CRISPR-Cas9 is an efficient system to knock out PD-1 from CTLs and PD-1 KO could significantly enhance the anti-tumor activity of CTLs.

14.
Arch Biochem Biophys ; 640: 53-60, 2018 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-29325758

RESUMO

MicroRNA-186-5p (miR-186-5p) is upregulated and exhibits as a crucial oncogene in various human tumors. However, the functions and underlying mechanisms of this microRNA on colorectal cancer remain largely unknown. Here, we report that miR-186-5p share a lower expression in colorectal cancer cell lines (HT116, H29, SW620 and LoVo) than in normal colonic epithelial cell line NCM460. MiR-186-5p overexpression inhibits proliferation, metastasis and epithelial-to-mesenchymal transition (EMT) of colorectal cancer cell line LoVo. Zinc Finger E-Box Binding Homeobox 1 (ZEB1), an EMT related marker, is predicted as a target of miR-186-5p. Luciferase reporter assay, qRT-PCR and western blot analysis showed that miR-186-5p directly targeted the 3'-untranslated regions (3'UTR) of ZEB1 messenger RNA. Further functional experiments indicated that overexpression of miR-186-5p suppress the proliferation and metastasis ability of LoVo, which was consistent with the inhibitory effects by knockdown of ZEB1. Additionally, overexpression of ZEB1 could significantly reverse the miR-186-5p mimics initiated suppression impact of proliferation, metastasis and EMT on LoVo. In summary, miRNA-186-5p affects the proliferation, metastasis and EMT process of colorectal cancer cell by inhibition of ZEB1. Hence, it may serve as a promising therapeutic target for colorectal cancer.


Assuntos
Proliferação de Células/fisiologia , Neoplasias Colorretais/patologia , Transição Epitelial-Mesenquimal/fisiologia , MicroRNAs/fisiologia , Regulação para Cima , Homeobox 1 de Ligação a E-box em Dedo de Zinco/metabolismo , Animais , Linhagem Celular Tumoral , Neoplasias Colorretais/metabolismo , Regulação para Baixo , Xenoenxertos , Humanos , Camundongos , Camundongos Nus , Metástase Neoplásica , RNA Mensageiro/genética , Homeobox 1 de Ligação a E-box em Dedo de Zinco/genética
15.
Inflammation ; 41(2): 667-676, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29264744

RESUMO

To investigate the potential role of interleukin-18 (IL-18) in immunomodulation during tumorigenesis of esophageal carcinoma and elucidate the underlying molecular mechanism, we employed IL-18 knockout mice for this purpose. Carcinogen 4-nitroquinoline 1-oxide (4NQO) was administrated in drinking water to induce occurrence of esophageal squamous cell carcinoma (ESCC). T cell activation as indicated by the surface CD molecules was analyzed with flow cytometry. The serous content of interferon-γ (IFN-γ) along with other cytokines was determined by inflammatory human cytokine cytometric bead array. The cytotoxicity assay was performed by co-culture of tumor cells with immune cells and relative cell viability was determined by lactate dehydrogenase (LDH) assay. Apoptotic cells were stained with Annexin-V/propidium iodide (PI) and analyzed by flow cytometry. Cell proliferation was measured with Cell Counting Kit-8 (CCK-8) assay. Our data demonstrated that deficiency of IL-18 promoted the progression and development of 4NQO-induced ESCC. Loss of IL-18 suppressed the activation of T cells in the esophagus. Deficiency of IL-18 inhibited the IFN-γ production by CD8+ T cells and natural killer (NK) cells. Absence of IL-18 inhibited the cytotoxicity of CD8+ T cells and NK cell in vitro. Moreover, deficiency of IL-18 promoted the apoptosis of CD8+ T cells and inhibited the proliferation of CD8+ T cells in vitro. Our data elucidated the immunomodulatory role of IL-18 during tumorigenesis of ESCC, whose deficiency compromised antitumor immunity and contributed to immune escape of esophageal carcinoma. Our results also indicated the therapeutic potential of exogenous IL-18 against ESCC, which warrants further investigations.


Assuntos
Linfócitos T CD8-Positivos/metabolismo , Neoplasias Esofágicas/etiologia , Interferon gama/antagonistas & inibidores , Interleucina-18/deficiência , Células Matadoras Naturais/metabolismo , Animais , Linfócitos T CD8-Positivos/imunologia , Carcinoma de Células Escamosas/induzido quimicamente , Carcinoma de Células Escamosas/etiologia , Carcinoma de Células Escamosas/imunologia , Neoplasias Esofágicas/imunologia , Humanos , Interferon gama/biossíntese , Células Matadoras Naturais/imunologia , Camundongos , Camundongos Knockout
16.
Gene ; 636: 36-41, 2017 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-28888577

RESUMO

T cell-mediated anti-tumor immunity plays a pivotal role in cancer immune surveillance. Cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) is a protein receptor mainly expressed in activated T cells and regulatory T cells. CTLA-4 competes with CD28 for ligand binding and generates inhibitory signals to attenuate T cell activation. The blockade of CTLA-4 mediated immune inhibitory checkpoint has been associated with enhanced anti-tumor immunity. In this study, we use CRISPR-Cas9 system to knock out (KO) CTLA-4 from cytotoxic T lymphocytes (CTLs) and evaluate its effect on the anti-tumor activity of the CTLs. CTLA-4 KO CTLs robustly enhanced tumor cell death by 40% compared to the control and facilitated apoptosis and caspase activities in tumor cells. The knockout of CTLA-4 also increased TNF-α and IFN-γ secretion of the CTLs by approximately 2-fold. The effectiveness of CTLA-4 KO in enhancing anti-tumor activity of the CTLs was verified in vivo using mouse xenograft model. The xenografted mice treated with CTLA-4 KO CTLs demonstrated repressed tumor growth and prolonged survival compared to the control group. Our data suggest that CRISPR targeting CTLA-4 immune checkpoint could significantly improve the anti-tumor activity of CTLs.


Assuntos
Sistemas CRISPR-Cas , Antígeno CTLA-4/genética , Neoplasias/imunologia , Linfócitos T Citotóxicos/imunologia , Animais , Apoptose , Antígeno CTLA-4/antagonistas & inibidores , Caspases/metabolismo , Células Cultivadas , Células Matadoras Induzidas por Citocinas/classificação , Citocinas/biossíntese , Células Dendríticas/classificação , Células HCT116 , Humanos , Imunofenotipagem , Camundongos SCID , Neoplasias/patologia , Neoplasias/terapia , Linfócitos T Citotóxicos/classificação
17.
Int J Clin Oncol ; 22(4): 641-650, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28664300

RESUMO

Brachytherapy is an important radio-therapeutic modality for a variety of malignancies, including prostate cancer, cervix cancer, breast cancer, vagina cancer, endometrium cancer, head and neck cancer, and many more. This technique has been shown to be an effective and safe non-pharmaceutical treatment with fewer serious complications and better outcome than other treatments for breast cancer. Every year, hundreds of thousands of patients around the world benefit from brachytherapy, which reliably delivers a relatively higher radiation dose to the intended target. However, the follow-up time, patient eligibility criteria, treatment strategy, and radiation doses used in published studies are somewhat inconsistent, making it difficult to strictly compare and evaluate the performance of the treatment. More rigorous studies are required to confirm the safety of this technique and to make outcome data more comparable. In this review, we focus on recent advances in breast brachytherapy techniques and provide an overview of outcomes, cosmetic outcome, toxicity, complications, and limitations of brachytherapy for the treatment of breast cancer. We also summarize the clinical outcomes and toxicity results in patients receiving or not receiving brachytherapy.


Assuntos
Braquiterapia/efeitos adversos , Braquiterapia/métodos , Neoplasias da Mama/radioterapia , Braquiterapia/instrumentação , Neoplasias da Mama/patologia , Feminino , Humanos , Dosagem Radioterapêutica , Resultado do Tratamento
18.
J Biomed Nanotechnol ; 13(3): 290-302, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29381284

RESUMO

In this paper, poly(lactic-co-glycolic acid) (PLGA) was used to fabricate spinal cord scaffolds using low temperature deposition manufacturing (LDM) technology. The PLGA scaffolds were characterized as having good porosity, hydrophilicity and considerable biodegradability. The effects of the PLGA scaffolds on cell proliferation and cytotoxicity were evaluated by culturing Schwann cells (SCs) on the surfaces of the scaffolds. The results showed that the SCs spread and proliferated well on the PLGA scaffolds. Histological assessment including Glia fibrillary acidic protein (GFAP) staining, Nissl staining, Luxol fast blue (LFB) staining and Bielschowsky silver staining showed that the spinal cord recoveries considerably improved with the PLGA scaffolds, indicating that the PLGA scaffolds exhibited potential for applications in the management of spinal cord injuries.


Assuntos
Regeneração Tecidual Guiada/instrumentação , Ácido Láctico/química , Ácido Poliglicólico/química , Células de Schwann/transplante , Traumatismos da Medula Espinal/terapia , Regeneração da Medula Espinal/fisiologia , Tecidos Suporte , Animais , Desenho de Equipamento , Análise de Falha de Equipamento , Feminino , Masculino , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Impressão Tridimensional , Ratos , Ratos Sprague-Dawley , Células de Schwann/citologia , Traumatismos da Medula Espinal/patologia , Engenharia Tecidual/instrumentação , Resultado do Tratamento
19.
Radiat Res ; 186(4): 377-384, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27643877

RESUMO

To elucidate the role of the mismatch repair gene Mlh1 in genome instability during the fetal stage, spontaneous mutations were studied in Mlh1-deficient lacZ-transgenic mouse fetuses. Mutation levels were high at 9.5 days post coitum (dpc) and gradually increased during the embryonic stage, after which they remained unchanged. In addition, mutations that were found in brain, liver, spleen, small intestine and thymus showed similar levels and no statistically significant difference was found. The molecular nature of mutations at 12.5 dpc in fetuses of Mlh1+/+ and Mlh1-/- mice showed their own unique spectra, suggesting that deletion mutations were the main causes in the deficiency of the Mlh1 gene. Of note, fetuses of irradiated mice exhibited marked differences such as post-implantation loss and Mendelian distribution. Collectively, these results strongly suggest that high mutation ofMlh1-/--deficient fetuses has little effect on the fetuses during their early developmental stages, whereas Mlh1-/--deficient fetuses from X-ray irradiated mothers are clearly effected.


Assuntos
Desenvolvimento Embrionário/genética , Proteína 1 Homóloga a MutL/deficiência , Taxa de Mutação , Animais , Animais Recém-Nascidos , Reparo de Erro de Pareamento de DNA/genética , Reparo de Erro de Pareamento de DNA/efeitos da radiação , Desenvolvimento Embrionário/efeitos da radiação , Feto/embriologia , Feto/efeitos da radiação , Instabilidade Genômica/genética , Instabilidade Genômica/efeitos da radiação , Genótipo , Camundongos , Proteína 1 Homóloga a MutL/metabolismo , Raios X/efeitos adversos
20.
Mar Drugs ; 14(7)2016 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-27367705

RESUMO

The purified total sterols and ß-sitosterol extracted from Sargassum horneri were evaluated for their antidepressant-like activity using the forced swim test (FST) and tail suspension test (TST) in mice. Total sterols and ß-sitosterol significantly reduced the immobility time in the FST and TST. Total sterols were administered orally for 7 days at doses of 50, 100, and 200 mg/kg, and ß-sitosterol was administered intraperitoneally at doses of 10, 20, and 30 mg/kg. ß-sitosterol had no effect on locomotor activity in the open field test. In addition, total sterols and ß-sitosterol significantly increased NE, 5-HT, and the metabolite 5-HIAA in the mouse brain, suggesting that the antidepressant-like activity may be mediated through these neurotransmitters.


Assuntos
Antidepressivos/química , Antidepressivos/farmacologia , Sargassum/química , Sitosteroides/química , Sitosteroides/farmacologia , Esteróis/química , Esteróis/farmacologia , Animais , Comportamento Animal/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Depressão/tratamento farmacológico , Elevação dos Membros Posteriores/métodos , Ácido Hidroxi-Indolacético/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Atividade Motora/efeitos dos fármacos , Neurotransmissores/metabolismo , Serotonina/metabolismo , Natação/fisiologia
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