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AIM and METHODS:The Ph.D.-7 phage display library was used to isolate peptides specific for glioma SWO-38 cell by whole cell screening.Moreover,binding efficiency analysis was carried out to test the binding specificity of the clones obtained. RESULTS: After three rounds of biopanning,a high concentration of phage clones was obtained and two of them were found to be highly specific to glioma SWO-38. CONCLUSION: Highly specific clones against neurtral glioma cells can be obtained from a phage display library by simple procedures.
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It has long been acknowledged that there is a link between inflammation and cancer, but its molecular mechanism remains unclear. A key player in inflammation is nuclear transcription factor NF-?B, that activity is triggered in response to infectious agents and proinflammatory cytokines via the I?B kinase. In parenchyma cells, inflammation through I?B kinase/NF-?B pathway suppresses apoptosis, accelerates cell cycle, then promote tumorigenesis. In mesenchyma cells inflammation through I?B kinase/NF-?B pathway produces cytokines and chemokines that may serve as tumor growth factors. To sum up, I?B kinase/NF -?B pathway represents a critical molecular link between inflammation and cancer.
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AIM: To isolate peptides targeted binding and internalizing into glioblastoma cell line SWO-38. METHODS: Tumor cells were screened five rounds of whole cell screen through the Ph.D.-12 phage display library. The monoclone specific binding efficiency to the tumor cell was analyzed, and the DNA of phages were extracted, sequenced and translated to the sequences of amino acid. RESULTS: In the phage library after five rounds of screen , 10 of 13 monoclones had highly selective binding to SWO-38 cells. We found two repeated peptide sequences. CONCLUSION: Whole cell screening against tumor cells through random phage peptide library can obtain phage peptides with highly specific binding and internalizing ability. The peptides could be used as a therapy vector for tumor targeted delivery.
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AIM: Three different antisense oligonucleotides complementary to basic fibroblast growth factor (bFGF) mRNA were compared in inhibitory effect on gene targeted expression. METHODS: After transfecting bFGF antisense oligonucleotides (asODN) into SWO-38 cells by lipofectin,the proliferation of cells was identified by MTT method, apoptosis was examined by flow cytometric cell cycle analysis and the expression levers of bFGF were detected by Western-blotting. RESULTS: There were 49%,33%,51% inhibition of cell growth and 35%, 27%, 18% cell apoptosis after asODN1, asODN2 and asODN3 treatment.In addition,the decrease in bFGF protein was 63%, 42%, 11%, respectively. CONCLUSION: The data suggeste that asODN1 is a potent target to bFGF mRNA, which inhibits cell growth and induces apoptosis in SWO-38 cells.
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Aim To investigate the role of p38 mitogen-activated protein kinases(MAPKs) in genistein-induced osteoblastic differentiation of mouse bone marrow-derived mesenchymal stem cells(BMSCs).Methods Mouse BMSCs cultured in phenol red-free ?-MEM containing 10% V/V FBS,were added ?-glycerophosphate and ascorbic acid for inducing osteoblastic differentiation,and treated with 0.01~1 ?mol?L~(-1) genistein and/or SB203580 and PD98059.The temporal sequence of osteoblastic differentiation in BMSCs cultures was assayed by measuring alkaline phosphatase activity(ALP) and calcium deposition.The MAPK phosphorylation level was detected by Western-blot.Results Genistein(0.01~1 ?mol?L~(-1)) showed a dose-dependent effect on osteoblastic differentiation as evidenced by increased alkaline phosphatase(ALP) activity and calcium deposition in mouse BMSCs cultures.Genistein(1 ?mol?L~(-1))-induced osteoblastic differentiation of BMSCs was diminished by p38 MAPK inhibitor but not the p44/42 MAPK inhibitor.The effects of Genistein were associated with rapid and sustained activation of p38 MAPK in the BMSCs cultures,which was blocked by SB203580(1 ?mol?L~(-1)).In contrast,Genistein treatment resulted in inactivation of p42/44MAPK,which was further attenuated by PD98059(25 ?mol?L~(-1)).Conclusion p38 MAPK plays an important role in genistein-induced osteoblastic differentiation of mouse BMSCs cultures.
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AIM: To study the role of androgen receptor (AR) in hormone-dependent and hormone-independent prostate cancer cell proliferation by knocking down AR expression with adenovirus-delivered siRNA. METHODS: Four well-designed siRNAs were synthesized and inserted into the adenovirus plasmid pShuttle-H1-Ri. The recombinant pShuttle-H1-Ri-AR plasmid was then co-transfected with pcDNA-AR to HEK293 cell line and Western blot was used to detect the inhibitory efficiency of different siRNAs on AR expression. Recombinant adenovirus containing more efficient siRNAs were prepared and used to infect three different humane prostate cancer cell lines including LNCapC4-2B and CWR22Rv1. The efficiency of knocking down AR expression was detected by Western blot. The effect of AR-knocking down on cell proliferation was detected by MTT colorimetric assay. RESULTS: All of the four designed siRNAs could knock down AR expression in transient co-transfection. Infecting with recombinant adenovirus containing more efficient siRNAs in hormone-dependent and hormone-independent prostate cancer cell lines specifically knocked down AR expression with high efficiency. Knocking down AR expression significantly decreased the proliferation rate in all these prostate cancer cells. CONCLUSION: The suppressed expression of AR in prostate cell lines mediated by siRNA could efficiently inhibit the cell proliferation, and these results show that AR plays an important role in the proliferation of hormone-dependent and hormone-independent prostate cancer cells. AR is an important therapeutic target for the treatment of prostate cancer.
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Objective:To clarify the contribution of endogenous bFGF, bFGF mRNA and FGFR1 to the abnormal growth and phenotypic transformation of neoplastic tumors cells.Methods:The antisense oligonucleotide primers was used to evaluate the influence of endogenous bFGF on growth of human glioma malignant cell lines SWO-38 in vitro. MTT was used to examine the variety of cells growth treated with bFGF antisense oligonucleotide primers. The methods of ELISA, in situ hybridization, immuno-hischemistry and image analysis were used to detect the expression level of bFGF, bFGF mRNA and FGFR1. The colony formation of cells in soft agar was used to assess the cloning efficiency of the cells after exposed to bFGF antisense oligo-nucleotide primers.Results:The cells multiplication, expression of bFGF mRNA and FGFR1 was inhibited by bFGF antisense oligonucleotide primers,and the cells multiplication was dose-dependent. Treated with antisense oligo-nucleotide primers, the expression of FGFR1 and secretion of bFGF were distinctly reduced, and the inhibition efficiency of cells multiplication of WSO-38 was 48% and the inhibition efficiency of colonies of SWO-38 in soft agar was 35%. The inhibition of cells multiplication can be reversed completely by external bFGF, and the reverse efficiency was 8%.Conclusion:The synthesis of bFGF mRNA and expression of bFGF can be specifically inhibited by antisense oligonucleotide, but the inhibition can be cleared up with the addition of external bFGF. The study suggested that the bFGFantisense oligonucleotide could have good effect in inhibiting of tumor under special condition.