ABSTRACT
PURPOSE: Cancer-associated fibroblasts (CAFs) activated by cancer cells has a central role in development and malignant biological behavior in colorectal cancer (CRC). Adult fibroblasts do not express Snail, but Snail-positive fibroblasts are discovered in the stroma of malignant CRC and reported to be the key role to chemoresistance. However, the reciprocal effect of CAFs expressed Snail to chemoresistance on CRC cells and the underlying molecular mechanisms are not fully characterized. MATERIALS AND METHODS: Snail-overexpressed 3T3 stable cell lines were generated by lipidosome and CT26 mixed with 3T3-Snail subcutaneous transplanted CRC models were established by subcutaneous injection. Cell Counting Kit-8, flow cytometry and western blotting assays were performed, and immunohistochemistry staining was studied. The cytokines participated in chemoresistance was validated with reverse transcriptase-polymerase chain reaction and heatmap. RESULTS: Snail-expression fibroblasts are discovered in human and mouse spontaneous CRCs. Overexpression of Snail induces 3T3 fibroblasts transdifferentiation to CAFs. CT26 co-cultured with 3T3-Snail resisted the impairment from 5-fluorouracil and paclitaxel in vitro. The subcutaneous transplanted tumor models included 3T3-Snail cells develop without restrictions even after treating with 5-fluorouracil or paclitaxel. Moreover, these chemoresistant processes may be mediated by CCL1 secreted by Snail-expression fibroblasts via transforming growth factor β/nuclear factor-κB signaling pathways. CONCLUSION: Taken together, Snail-expressing 3T3 fibroblasts display CAFs properties that support 5-fluorouracil and paclitaxel chemoresistance in CRC via participation of CCL1 and suggest that inhibition of the Snail-expression fibroblasts in tumor may be a useful strategy to limit chemoresistance.
Subject(s)
Adult , Animals , Humans , Mice , Blotting, Western , Cell Count , Cell Line , Colorectal Neoplasms , Cytokines , Drug Resistance, Multiple , Fibroblasts , Flow Cytometry , Fluorouracil , Immunohistochemistry , In Vitro Techniques , Injections, Subcutaneous , Paclitaxel , Snails , Transforming Growth FactorsABSTRACT
To investigate the impact of phenotypic knockout of CXCR4 on Molt-4 cells via intrakine technology,the C-terminal alpha-helix gene SDF-1alpha/54/KDEL of human stromal cell-derived Faceor-1 deletion is fused to a retention signal 4-peptide -KDEL that retains the newly synthesized receptor within the Molt-4 cells endoplasimc reticulum. Subsequently, PCR is used to amplify the target gene SDF-1alpha/54/ KDEL from the constructed plasmid SDF-WT-Gly x 4-Dec/PET-30a(+) at its C-terminal and subclone it into eukaryotic expression vectors pEGFP-C3 for generating recombinant vector cells by lipEGFP-C3/SDF-1alpha/54/KDEL, and then have it sequenced. After the transfection of recombinant plasmids into COS-7 posome, SDF-1alpha/54/KDEL protein is confirmed with Western blot. The recombinant plasmids pEGFP-C3/SDF-1alpha/54/KDEL are isolated and transiently transfected in Molt-4 cells by electroporation. Flow cytometric analysis shows a dramatic reduction of CXCR4 expression on Molt-4 cells. The conclusion is that SDF-1alpha/54/KDEL could assume a role in the phenotypic knockout of CXCR4, and the findings suggest that the inhibiting effect of SDF-1alpha/54 against CXCR4 is not influenced by the deletion of SDF-1alpha helix at the C terminal.
Subject(s)
Animals , Humans , COS Cells , Cell Membrane , Metabolism , Chlorocebus aethiops , Chemokine CXCL12 , Genetics , Cloning, Molecular , Electroporation , Gene Knockout Techniques , Genetic Vectors , Genetics , Mutation , Receptors, CXCR4 , Genetics , Metabolism , Recombinant Proteins , Genetics , Stromal Cells , Metabolism , TransfectionABSTRACT
OBJECTIVE: To investigate the status quo of pharmacists training in Hong Kong,and to provide reference for pharmaceutical education reform of college in mainland.METHODS: The status quo,role,license examination and culture system of licensed pharmacist in Hong Kong were analyzed.Based on the practice of our university,the difference in culture system of pharmacentical talents and curriculum setting of pharmaceutical education were compared between Hong Kong and mainland.RESULTS & CONCLUSION: On the basis of education reform practice of our university for Hong Kong students,it is suggested to match the pharmacy curriculum setting and training program used in Hong Kong,to improve the clinical practice,to explore "4+2" culture model,and to enhance English training of pharmaceutical education.
ABSTRACT
Decorin (DCN) is a member of the small leucine-rich proteoglycan gene family. Many studies indicated that DCN inhibited fibrosis and scar-formation by neutralization of TGF-P and interfering the binding of TGF-beta with its receptor, which induced ectopic deposition of extracellular matrix. Additionally, DCN can prevent the proliferation and metastasis of tumor cells by activating EGFR/MAPK/p21 signal pathway and inhibiting the cell proliferation pathway mediated by EGF-EGFR. It is suggested that the recombinant DCN had potential pharmaceutical potency in treatment of chronic fibrosis and neoplasm for its critical biological activities and low immunogenicity.
Subject(s)
Animals , Humans , Antineoplastic Agents , Pharmacology , Decorin , Extracellular Matrix Proteins , Chemistry , Pharmacology , Fibrosis , Proteoglycans , Chemistry , Pharmacology , ErbB Receptors , Recombinant Proteins , Pharmacology , Transforming Growth Factor beta1ABSTRACT
The chemokine SDF-1 (CXCL12) and its receptor, CXCR4, have been implicated in organ-specific metastases of several malignancies. CXCR4 expression has recently been characterized in many cancer cell types and is thought to play a pivotal role in directing the migration of metastasizing tumor cells to SDF-1-rich tissues. SDF-1, which is highly expressed in the organs where breast cancers preferentially metastasize, has been shown to promote cancer cell migration. The tumor cells use chemotaxis which occurred between CXCR4 and its ligand SDF-1 to direct migration from their primary sites via the circulation to the preferential sites of metastases, and further studies on the mechanism involved in a variety of cellular signaling pathways are beneficial to tumor therapy.
Subject(s)
Humans , Breast Neoplasms , Pathology , Chemokine CXCL12 , Physiology , Multiple Myeloma , Pathology , Neoplasm Metastasis , Receptors, CXCR4 , Physiology , Signal Transduction , PhysiologyABSTRACT
It is no doubt that the gene therapy using recombinant engineering cells provides a novel approach to many refractory diseases. However, the transplant rejection from the host's immune system against heterogeneous cells has been the main handicap of its clinical application. The modern cell micro-encapsulation technique with good immune isolation makes it possible to overcome this problem and has shown potential application foreground in clinical therapies for a lot of diseases such as Parkinson's disease and Hemophiliac disease. This article reviews mainly the relative materials and techniques in processing micro-encapsulation, the host cells used to construct the recombinant genetic engineering cells and application of cell micro-encapsulation technique in the field of gene therapy.
Subject(s)
Humans , Biomedical Engineering , Methods , Cell Transplantation , Methods , Genetic Therapy , Miniaturization , Tissue Engineering , MethodsABSTRACT
The aim of this study is to explore the possibility and technical itinerary of establishing an mammal engineering cell line in which hBD-2 can be effectively expressed, secreted, detected, separated and purified. The full hBD2 cDNA was inserted into the multi clone site of a eukaryotic expressive plasmid pcDNA3.1/Myc-His(+) and located closely at the upstream of two tag gene (myc and 6 Poly-histidines) so as to construct another recombinant eukaryotic expressive vector of hBD-2 gene: rpcDNA3.1/Myc-His/hBD-2. By the use of RT-PCR with special primers, a band of 240 bp was amplified from COS-7 cells transfected by this recombinant plasmid, which matched full length of cDNA coding hBD2 plus myc epitope and 6 poly-histidines tags. Western blot analysis with specific anti-histidines antibody revealed that the lysate of COS-7 cells transfected by rpcDNA3.1/Myc-His/hBD-2 had a strong band with molecular weight of about 10 Kd that was approximate to the size of chiasmic peptide. Antibacterial activity assay showed that obvious bacterial inhibition occurred in both lysate and supernatant of COS-7 cells transfected by rpcDNA3.1/Myc-His/hBD-2.
Subject(s)
Animals , Humans , Base Sequence , Blotting, Western , COS Cells , Gene Expression , Genes, myc , Histidine , Genetics , Molecular Sequence Data , Plasmids , Reverse Transcriptase Polymerase Chain Reaction , Transfection , beta-Defensins , Genetics , PharmacologyABSTRACT
Aim To investigate whether SC58125 synergized with TNF-? to induce HT-29 cell apoptosis and study the possible molecular mechanism. Methods By using MTT, agarose gel electrophoresis and flow cytometry, we examined the effect of SC58125/TNF-? on cell proliferation and apoptosis in HT-29 cells. The activity of caspase-3 and the changes of I?B? and NF-?B were also measured after treatment with SC58125 by Electrophoretic mobility shift assay and Western blot. Results Both SC58125 and TNF-? exhibited cytotoxicity, the combination of the two agents significantly reduced HT-29 cell viability in a dose-dependent manner. TNF-?-treated cells showed oligonucleosomal cleavage of genomic DNA. SC58125 significantly enhanced the inhibition of cell proliferation and inducement of cell apoptosis of TNF-?,the apoptotic index was increased from 11.2%?1.1% to 53.9%?2.1%. SC58125/TNF-?-induced apoptosis of HT-29 cells was accompanied by the induction of caspases-3. I?B? levels were substantially decreased after treatment with TNF-? and the degradation of I?B? was almost completely inhibited when SC58125 was added in NF-?B was activated in HT-29 cells after treatment with TNF-?, whereas pretreatment of HT-29 cells with SC58125 for 2 h, TNF-?-induced NF-?B DNA binding was profoundly inhibited. Conclusion SC58125 synergizes with TNF-? to inhibit cell growth and induce apoptosis in HT-29 cells, which may be mediated by activating caspases and preventing degradation of I?B?.
ABSTRACT
Many snake venoms contain complex mixtures of pharmacologically important molecules, some of which show potential therapeutic value in the treatment of cancer and other human disorders. In this review, we mainly reports the effects of snake venom active components, such as disintegrins and lectins in paralyzing cancer cells, blocking on cell migration, interaction with integrins, inhibition of tumor dissemination and angiogenesis. The advanced researches on the snake venom's apoptosis-inducing components on tumors are also introduced. [