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Liu Q.,First Hospital of Handan City | Dong C.,First Hospital of Handan City | Li L.,Hebei Normal University | Sun J.,Hebei Medical University | Li C.,Hebei Normal University
Chinese Journal of Lung Cancer | Year: 2011

Background and objective: Survivin, a member of the inhibitor of apoptosis (IAP) protein family, has been demonstrated as a potential new target for apoptosis-based therapy in cancer and lymphoma. The aim of this study is to investigate effects and mechanisms of survivin siRNA transfection on lung adenocarcinoma cell lines SPCA1 and SH77. Methods: A siRNA plasmid expression vector and pSi scrambled against survivin were constructed and transfected into SPCA1 and SH77 cells with Lipofectamine 2000. The proliferations of lung adenocarcinoma SPCA1 and SH77 cells were detected by MTT. The apoptotic rate and cell cycle were detected by flow cytometer. The activity of survivin mRNA and protein expression were analyzed with RT-PCR and Western blot. Results: Survivin siRNA reduced the proliferation of SPCA1 and SH77 cells. Cell cycle was inhibited in G0/G1. Expressions of survivin siRNA mRNA and protein were reduced in transfected cells compared with the control cells. Conclusion: siRNA targeted against survivin can effectively suppress SPCA1 and SH77 cells proliferation and significantly induce SPCA1 and SH77 cells apoptosis. Source


Hu Z.,260th Hospital of PLA | Du Y.,260th Hospital of PLA | Liu Q.,First Hospital of Handan City | Wang Y.,Hebei Normal University | Li L.,Hebei Normal University
Chinese Journal of Lung Cancer | Year: 2010

Background and objective: The spider venom may inspire new drugs to treat cancer. The aim of this study is to investigate the effects and mechanisms of spider venom on lung adenocarcinoma cell A549. Methods: The proliferation of lung adenocarcinoma A549 cells was detected by MTT. The apoptosis rate was observed with MTT assay and flow cytometer. The activity of catalase was detected by colorimetry. The malondialdehyde (MDA) content was determined by improved thiobarbituric acid fluorometric method. The expression of P38MAPK protein was analyzed with Western blot. Results: Spider venom can remarkably inhibite the proliferation of lung adenocarcinoma A549 cells, increased activity of catalase and MDA content, down-regulated expression of P38MAPK compared with the control group. Conclusion: The reduced proliferation of lung adenocarcinoma A549 cells by spider venom is may be associated with the increased of activity of catalase and MDA content and decreased expression of P38MAPK. Source


Su Y.-H.,Hebei Medical University | Zhang S.-B.,Third Hospital of Shijiazhuang City | Cui H.-X.,Hebei Medical University | Kang L.,Hebei Medical University | And 3 more authors.
Acta Anatomica Sinica | Year: 2013

Objective: To examine the correlation of draxin expression and neural crest migration in chick embryonic spinal cord at different developmental stages. Methods: Ten chick embryos at different stages were used. In situ hybridization and immunohistochemistry were used to observe the localization of draxin expression, the migration pathway of neural crest cell-the spatial correlation of draxin expression and the neural crest migration. Living tissue incubation with draxin alkaline phosphatase (draxin-AP) recombinant protein was used to check the binding ability of migrating neural crest with draxin protein. Results: With development, the draxin expression and neural crest migration had a very similar anterior-posterior gradient in chick embryonic spinal cord. Draxin was expressed in the dorsal neural tube, roof plate and dorsal tip of dermomyotome which surrounded the migration pathway of neural crest cells. We also found that portion of migrating neural crest cells bound to draxin-AP protein directly. Conclusion: Draxin is expressed in the tissues surrounding the neural crest migration pathway and portion of migrating neural crest cells binds to draxin protein directly. We conclude therefore that draxin may be involved in the regulation of neural crest cell migration in chick embryonic spinal cord. Source


Nan C.,Hebei Medical University | Guo L.,Hebei Medical University | Zhao Z.,Hebei Medical University | Ma S.,Hebei Medical University | And 4 more authors.
International Journal of Oncology | Year: 2016

The present study evaluated the ability and optimal concentration of tetramethylpyrazine (TMP) to induce human umbilical cord-derived mesenchymal stem cells (hUMSCs) to differentiate into neuron-like cells in vitro. Human umbilical cords from full-term caesarean section patients were used to obtain hUMSCs by collagenase digestion after removal of the umbilical artery and vein. The surface antigen expression profile of cultured hUMSCs was monitored by flow cytometry. After amplification, cells of the 5th passage were divided into experimental groups A-C treated with TMP at 4.67, 2.34 and 1.17 mg/ml, respectively, in low glucose-Dulbecco's Modified Eagle's Medium (L-DMEM) (induction medium), while group D (control) was exposed to L-DMEM culture medium only. Differentiation of hUMSCs into neuron-like cells and morphological changes were observed every 0.5 h with an inverted phase contrast microscope for 6 h. After the 6-h induction period, proportions of cells expressing neuronal markers neuron-specific enolase (NSE), neuroflament protein (NF-H) and glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. The optimal concentration of TMP was selected on the basis of neuron-like cell positive rate. Western blotting and RT-polymerase chain reaction were applied to detect the expression of NSE, NF-H, and GFAP of the group of optimal concentration in each point-in-time. Results showed that most primary cells were adherent 12 h after seeding and first appeared as diamond or polygon shapes. Thereafter, they gradually grew into long spindle-shaped cells and finally in a radiating or swirling pattern. The cells maintained a strong proliferative capacity after continuous passage. Flow cytometry analysis of cultured hUMSCs at the 3rd, 5th and 10th passages expressed CD73, CD90 and CD105, but not CD11b, CD19, CD34, CD45 or human leukocyte antigen-DR. After 6 h of TMP treatment, typical neuron-like cells with many protrusions connected into a net-like pattern were observed in all experimental groups. These neuron-like cells were positive for NSE and NF-H, but negative for GFAP. Among the tested treatment groups, group A with TMP at 4.67 mg/ml had the highest expression of NSE and NF-H. By contrast, no change was found after induction in the control group. The mRNA expression of cells expressing neuronal markers as well as GAPDH was observed, with the relative NSE transcript levels of 0, 1.303±0.031, 1.558±0.025, 1.927±0.019 and 2.415±0.033 after 0, 1, 2, 4 and 6 h of treatment, respectively; the mRNA expression of NH-F was 0, 1.429±0.025, 1.551±0.024, 1.930±0.042 and 1.398±0.014 after 0, 1, 2, 4 and 6 h of treatment, respectively. There was no expression of GFAP before or after induction and all the groups showed high expression of GAPDH at each time point. Protein expression was also observed on cells expressing neuronal markers as well as GAPDH at each time point. The protein expression of NSE was 0, 0.717±0.097, 0.919±0.056, 1.097±0.143 and 1.157±0.055 in proper order; the protein expression of NH-F was 0, 0.780±0.103, 0.973±0.150, 1.053±0.107 and 0.753±0.094 in proper order. There was no expression of GFAP before or after induction, and all the groups showed high expression of GAPDH at each tested time point. Our results demonstrated that TMP can induce hUMSCs to differentiate into neuron-like cells effectively with the optimal concentration of 4.67 mg/ml. After induction, the NSE and NF-H of the neuron-like cells were positive, but the GFAP-2 was negative. Source


Nan C.,Hebei Medical University | Shi Y.,Hebei Medical University | Zhao Z.,Hebei Medical University | Ma S.,Hebei Medical University | And 4 more authors.
International Journal of Molecular Medicine | Year: 2015

In the present study, human umbilical cord-derived mesenchymal stem cells (hUMSCs) were investigated for their potential to be induced to differentiate in vitro into neuron-like cells by monosialoteterahexosyl ganglioside (GM1). Mononuclear cells obtained from umbilical cords from women with full-term pregnancies whose babies were delivered by cesarean section were cultivated in vitro and their surface antigen expression profiles were monitored. Following amplification, the cells were divided into 5 groups, of which 4 (groups A-D) were treated with GM1 at doses of 50, 100, 150 and 200 μg/ml, respectively. The control (group E) was treated with the vehicle only. The ability of GM1 to induce the differentiation of the hUMSCs into neuron-like cells was monitored for 6 h. The expression levels of microtubule-associated protein-2 (MAP-2), neurofilament protein (NF-H) and glial fibrillary acidic protein (GFAP) were measured by immunohistochemistry. Following exposure to GM1, the hUMSCs first appeared to have a diamond or polygonal shape and gradually grew into long spindle-shaped cells, finally exhibiting a radiating or swirling pattern. The cells maintained a strong proliferative capacity after continuous passage. Flow cytometry revealed that the hUMSCs expressed CD73, CD90 and CD105 up to passage 10, but not CD11b, CD19, CD34, CD45 or HLA-DR. Treatment with GM1 for 6 h led to the appearance of neuron-like cells with oval-shaped cell bodies and protruding neurites. These neuron-like cells were positive for MAP-2 and NF-H, but negative for GFAP expression. No changes in the expression of these markers were observed in the control group. Thus, the findings of the present study demonstrate that GM1 effectively induces hUMSCs to differentiate into neuron-like cells. Source

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