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Negin P Martin, Page Myers, Eugenia Goulding, Shih-Heng Chen, Mitzie Walker, Thomas M Porter, Lucas Van Gorder, Amanda Mathew, Artiom Gruzdev, Charles Romeo
Lentiviruses are highly efficient vehicles for delivering genes into cells. They readily transduce primary and immortalized cells in vivo and in vitro. Genes delivered by lentiviruses are incorporated and replicated as part of their host genome and therefore offer a powerful tool for creation of stable cell lines and transgenic animals. However, the zona pellucida surrounding the fertilized eggs acts as a barrier and hinders lentiviral transduction of embryos. Here, we utilize a laser, typically used to perforate the zona pellucida for in vitro fertilization, to permeabilize the zona for lentiviral gene delivery...
February 13, 2018: Transgenic Research
Gang Sun, Xin Sui, Dayue Han, Jianjun Gao, Yining Liu, Lin Zhou
The human tripartite motif (TRIM) 59 has been implicated in tumorigenesis of many types of cancer. However, the biological function and molecular mechanism of TRIM59 in hepatocellular carcinoma (HCC) remains unknown. In our study, the purpose was to investigate the impact of TRIM59 on the biologic behavior of HCC cells. We observed that TRIM59 was highly expressed in HCC cells compared with a normal human hepatocyte cell line. Lentivirus-mediated knocking down of TRIM59 significantly suppressed the proliferation, migration and invasion of HCC cells, whereas overexpression of TRIM59 enhanced cell growth and metastasis...
November 1, 2017: Die Pharmazie
Weihong Zhu, Xinzhan Mao, Wanchun Wang, You Chen, Ding Li, Hui Li, Pengcheng Dou
Cisplatin is an effective chemotherapeutic agent for osteosarcoma (OS) and has been shown to induce endoplasmic reticulum (ER) stress-associated apoptosis in human cancer cells. Ganglioside GD2-specific antibodies can inhibit tumor cell viability without involvement of the immune system. A recent study has shown that antiGD2 monoclonal antibody (mAb) 14G2a effectively inhibits the viability and invasiveness of human OS cells. In this study, we explored the effect of anti-GD2 mAb and cisplatin alone and in combination on ER stress-associated apoptosis in osteosarcoma cells...
February 1, 2018: Die Pharmazie
Yongwei Li, Zhengfei Shan, Bin Yang, Diandong Yang, Changping Men, Yuanshan Cui, Jitao Wu
AIMS: Adipose-derived stem cells (ADSCs), a source of mesenchymal stem cells, are able to differentiate into numerous cell lineages, including epithelial and smooth muscle cells. The use of ADSCs in tissue engineering technology has become the most promising therapeutic approach for urethral reconstruction. This study aimed to explore the effect of lncRNA highly upregulated in liver cancer (HULC) on the induction of ADSCs to differentiate into epithelial and smooth-muscle-like cells. METHODS: ADSCs were isolated from a male dog, and the expression of HULC in ADSCs was overexpressed by transfection with HULC expressing vector lentivirus...
January 2, 2018: Die Pharmazie
Jinqiu Wei, Yujiao Zhang, Zhan Li, Ximin Wang, Linlin Chen, Juanjuan Du, Jing Liu, Ju Liu, Yinglong Hou
BACKGROUND/AIMS: Cardiac autonomic nerve remodeling (ANR) is an important mechanism of atrial fibrillation (AF).GTP cyclohydrolase I (GTPCH-1), encoded by GCH1, is the rate-limiting enzyme in de novosynthesis of tetrahydrobiopterin (BH4), an essential cofactor for nitric oxide (NO) synthesis. Previous studies reported that increased BH4 and NO content negatively regulated nerve regeneration. This study investigated effects of GCH1on ANR via BH4 pathway, regulated by microRNA-206 (miR-206)...
February 13, 2018: Pacing and Clinical Electrophysiology: PACE
Guokai Dong, Man Wang, Guangfu Gu, Shanshan Li, Xiaoming Sun, Zhouru Li, Hongxing Cai, Zhengqiu Zhu
Metastasis-associsated in colon cancer 1 (MACC1), a newly identified oncogene, promotes tumor cell proliferation and invasion. In the present study, the expression of MACC1, hepatocyte growth factor (HGF) and its receptor, MET proto-oncogene (c-Met), was investigated in human gastric cancer tissues and adjacent normal tissues by immunohistochemistry. The association between the expression levels of the proteins and the clinicopathological parameters of the tumors were statistically analyzed. Furthermore, lentiviral particles expressing MACC1 were used to infect the hepatic satellite cell (HSC) line LX2...
March 2018: Oncology Letters
Huan Xu, Wenzhi Li, Kai Qi, Juan Zhou, Meng Gu, Zhong Wang
Previous studies have assessed nucleobindin 2 (NUCB2) expression in multiple urological cancer cell lines and detected its effect on renal cancer cell apoptosis. Additionally, certain reports have indicated a novel function of NUCB2 in promoting invasion in renal cancer. The levels of NUCB2 expression in different tumor cell lines were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Human NUCB2 and β-actin (ACTB) cDNA plasmids were inserted into lentivirus plasmids, which were then transfected into 786-O cells...
February 2018: Oncology Letters
Jibo Wang, Yingjie Zhao, Miaomiao Xin, Lin Pan, Liqin Wang, Kun Yang
The aim of the present study was to identify the effectiveness of lentivirus-mediated RNA interference (RNAi) targeting mouse tumor necrosis factor-α (TNF-α). RNAi lentivirus was used in vitro to transfect RAW264.7 cells, and the expression of TNF-α, interleukin (IL)-1β and IL-6 mRNAs and TNF-α protein in RAW264.7 cells was measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay, respectively. In vivo, mice with collagen-induced arthritis (CIA) were injected intravenously with RNAi lentivirus, and CIA arthritis scores and the serum levels of TNF-α were detected...
February 2018: Experimental and Therapeutic Medicine
Jian Zhang, Li Wang, Yongkang Zhang
NIMA-related kinase-7 (Nek7) is a centrosomal kinase involved in various types of cancer, including gallbladder cancer and hepatocellular carcinoma. However, the biological function and the potential underlying mechanism of Nek7 in retinoblastoma remain largely unknown. Therefore, the present study investigated the effects of Nek7 in retinoblastoma cells. The expression of Nek7 was initially determined and observed to be commonly upregulated in retinoblastoma cell lines (Y79, SO-RB50 and WERI-RB1) as compared with that in normal retinal pigment epithelium cells...
February 2018: Experimental and Therapeutic Medicine
Xi Zeng, Ping Huang, Mingkai Chen, Shiqian Liu, Nannan Wu, Fang Wang, Jing Zhang
The aim of the present study was to elucidate the effect of transmembrane protein 16A (TMEM16A) on portal vein smooth muscle cell (PVSMC) proliferation associated with portal vein remodeling in portal hypertension (PHT). Sprague-Dawley rats were subjected to bile duct ligation to establish a rat model of liver cirrhosis and PHT. Sham-operated animals served as controls. At 8 weeks after bile duct ligation, the extent of liver fibrosis and the portal vein wall thickness were assessed using hematoxylin-eosin staining...
January 2018: Experimental and Therapeutic Medicine
Ye He, Hai-Bo Li, Xin Li, Yi Zhou, Xiao-Bo Xia, Wei-Tao Song
BACKGROUND/AIMS: Retinal Müller cells could be induced to differentiate into retinal ganglion cells (RGCs), but RGCs derived from Müller cells have defects in axon growth, leading to a defect in signal conduction. In this study we aimed to explore the role of miR-124 in axon growth of RGCs derived from Müller cells. METHODS: Müller cells were isolated from rat retina and induced to dedifferentiate into retinal stem cells. The stem cells were infected by PGC-FU-Atoh7-GFP lentivirus and then transfected with miR-124 or anti-miR-124, and the length of axon was compared...
February 5, 2018: Cellular Physiology and Biochemistry
Yang Wang, Juan Yu, Lvjun Liu, Wen Li, Xingxiang Duan, Yingying Peng, Sicong Zeng, Qi Ouyang, Guangxiu Lu, Ge Lin, Yi Sun
The human embryonic stem cell (hESC) line NERCe003-A-1 was generated by introducing lentiviral-vector-mediated tetracycline-inducible β-catenin expression into a normal hESC line, NERCe003-A. The resulting cell line can overexpress the β-catenin protein, encoded by the CTNNB1 gene, after exposure to doxycycline (Dox). CTNNB1 gene expression was confirmed by quantitative PCR (qPCR) and immunofluorescence assays. Further characterization confirmed that the NERCe003-A-1 cell line expresses typical pluripotency markers and has the ability to form the three germ layers both in vitro and in vivo...
February 3, 2018: Stem Cell Research
Wei Hu, Xing-Bo Dang, Gang Wang, Shuai Li, Yue-Lin Zhang
Peroxiredoxins (PRDXs) are a highly conserved family of thiol peroxidases that scavenge peroxides in cells. PRDX3 is one member of PRDXs localized in the mitochondria, and has been shown to be involved in antioxidant defense and redox signaling. In this study, we investigated the role of PRDX3 in neuronal trauma using a traumatic neuronal injury (TNI) model in primary cultured cortical neurons. We found that TNI significantly decreased the expression of PRDX3 at both mRNA and protein levels. Overexpression of PRDX3 by lentivirus (LV-PRDX3) transfection attenuated lactate dehydrogenase (LDH) release and neuronal apoptosis after TNI...
February 7, 2018: Neurochemistry International
Caroline Le Duigou, Etienne Savary, Mélanie Morin-Brureau, Daniel Gomez-Dominguez, André Sobczyk, Farah Chali, Giampaolo Milior, Larissa Kraus, Jochen C Meier, Dimitri M Kullmann, Bertrand Mathon, Liset Menendez de la Prida, Georg Dorfmuller, Johan Pallud, Emmanuel Eugène, Stéphane Clemenceau, Richard Miles
BACKGROUND: Insights into human brain diseases may emerge from tissue obtained after operations on patients. However techniques requiring transduction of transgenes carried by viral vectors cannot be applied to acute human tissue. NEW METHOD: We show that organotypic culture techniques can be used to maintain tissue from patients with three different neurological syndromes for several weeks in vitro. Optimized viral vector techniques and promoters for transgene expression are described...
February 7, 2018: Journal of Neuroscience Methods
Jing Lin, Aihui Ji, Guanzhong Qiu, Huaizhi Feng, Jian Li, Shuo Li, Yongxiang Zou, Yong Cui, Chaoli Song, Hua He, Yicheng Lu
F-box and WD repeat domain-containing 7(FBW7) is a SCF-type E3 ubiquitin ligase targeting a multitude of oncoproteins for degradation. Acting as one of the most important tumor suppressor it is frequently inactivated in various tumors. In this study we aimed to evaluate the relationship of FBW7 with glioma pathology and prognosis, and examine its effect in glioma malignancies and temozolomide(TMZ)-based therapy. Clinical tissues and TCGA database analysis revealed FBW7 expression was correlated inversely with glioma histology and positively with patient survival time...
February 10, 2018: Cancer Science
Hai Liu, Xuanxuan Wang, Aihua Huang, Huaping Gao, Yikan Sun, Tingting Jiang, Liming Shi, Xianjie Wu, Qinghua Dong, Xiaonan Sun
Artemis is a key protein of NHEJ (non-homologous end-joining), which is the major pathway for the repair of IR-inducedDSBs in mammalian cells. However, it has not been largely investigated about the expression of Artemis in tumor and the influence of silencing Artemis on the tumor's sensitivity to radiation. In this study, we investigated how expression levels of Artemis may affect treatment outcome of radiotherapy and chemotherapy in colorectal cancer cells. Firstly, we found that the expression of Artemis is strong in some human rectal cancer samples, even can be higher than adjacent normal tissues using immunohistochemical staining...
February 9, 2018: Oncology Research
Valerie T Y Tan, Bruce G Mockett, Shane M Ohline, Karen D Parfitt, Hollie E Wicky, Katie Peppercorn, Lucia Schoderboeck, Mohamad Fairuz Bin Yahaya, Warren P Tate, Stephanie M Hughes, Wickliffe C Abraham
Alzheimer's disease (AD) is a neurodegenerative disease driven in large part by accumulated deposits in the brain of the amyloid precursor protein (APP) cleavage product amyloid-β peptide (Aβ). However, AD is also characterised by reductions in secreted amyloid precursor protein-alpha (sAPPα), an alternative cleavage product of APP. In contrast to the neurotoxicity of accumulated Αβ, sAPPα has many neuroprotective and neurotrophic properties. Increasing sAPPα levels has the potential to serve as a therapeutic treatment that mitigates the effects of Aβ and rescue cognitive function...
February 9, 2018: Molecular Brain
Q Zhang, N Yu, B-T Yu
OBJECTIVE: To investigate the role and mechanism of micro ribonucleic acid (miR)-298 in myocardial apoptosis after myocardial infarction. MATERIALS AND METHODS: In vivo experiments, the rat model of myocardial infarction was established, and miR-298 was up-regulated via lentivirus with miR-298 overexpression. Cardiac function of rats was detected via echocardiography, Bcl-2 associated X protein (BAX) expressions in infarction border zone were detected via Real-time Quantitative PCR (qT-PCR) and Western blot, and TUNEL assay was used to detect the myocardial apoptosis...
January 2018: European Review for Medical and Pharmacological Sciences
J Pei, P Li, Z-Y Zhang, H-L Zhang, Y-H Gao, D-Y Wang, Y Zheng, X Xu, J-Z Cui
OBJECTIVE: To explore the role of transgelin-2 TAGLN2 in the development and progression of meningioma and the potential regulatory. MATERIALS AND METHODS: TAGLN2 knockdown expression and overexpression in vitro models were constructed using lentivirus in meningioma cell line CH157; their corresponding transfection efficiencies were verified by qRT-PCR and Western Blot. Actions of TAGLN2 on the proliferation of meningioma cells were explored by CCK8 and colony formation assays...
January 2018: European Review for Medical and Pharmacological Sciences
Jamie L Everman, Cydney Rios, Max A Seibold
The adaptation of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR associated endonuclease 9 (CRISPR-Cas9) machinery from prokaryotic organisms has resulted in a gene editing system that is highly versatile, easily constructed, and can be leveraged to generate human cells knocked out (KO) for a specific gene. While standard transfection techniques can be used for the introduction of CRISPR-Cas9 expression cassettes to many cell types, delivery by this method is not efficient in many primary cell types, including primary human airway epithelial cells (AECs)...
2018: Methods in Molecular Biology
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