ABSTRACT
Objective@#To explore the mechanism of llinc00052 regulating Wnt pathway affecting invasion and metastasis of gastric cancer.@*Methods@#SGC-7901 cells were selected from gastric cancer cell lines, and siRNAs related to INC0005 and had invasion and metastasis effects in gastric cancer cells were screened by binding of IncRNA expression profiles to qRT-PCR.Ilinc00052 and miRNA expression changes were studied by in vitro cell transfection experiments.Through molecular experiments, the expression of llinc00052 and the effect on Wnt/β-catenin expression were investigated to explore whether llinc00052 could affect the invasion and metastasis of gastric cancer cells by regulating miRNAs affecting Wnt/β-catenin signaling pathway.@*Results@#Transwell chamber test showed that the number of transmembrane cells in the untransfected plasmid group was (134.10±4.29), and the number of transmembrane cells in the overexpressed llinc000522 plasmid group was (169.24±6.99)(t=8.956, P=0.001). The scratch test showed that the migration distance in the llinc000522 overexpression transfection plasmid group was significantly higher than that in the no-load plasmid transfection group(r=0.907, P<0.01). The clone formation rate of llinc000522 overexpressed transfected plasmid group was significantly higher than that of the empty plasmid group[(92.75±6.32)% vs.(73.34±9.14)%](t=5.998, P<0.05). Compared with the transfection of blank plasmid, the expressions of Wnt1, Wnt3a, Wnt2 and β-catenin mRNA in the llinc000522 overexpression transfection group were significantly up-regulated(P<0.05), while the miRNA transfection group had no significant effect on the expression.The expressions of Wnt1, Wnt3a, Wnt2, and β-catenin mRNA were significantly increased [(1.82±0.11), (1.52±0.15), (1.42±0.21), (1.71±0.19)](P<0.05), but their expressions were still lower than those of the genes transfected with llinc000522 alone.Compared with the blank plasmid transfection, the expressions of Wnt1, Wnt3a, Wnt2 and β-catenin protein in the llinc000522 overexpression transfection group were significantly up-regulated(all P<0.05), while the miRNA transfection group had no significant effect on its expression.The protein expressions of Wnt1, Wnt3a, Wnt2 and β-catenin were also significantly increased[(1.53±0.09), (1.4±0.21), (1.33±0.07), (1.47±0.19)](P<0.05), but their expressions were still lower than those of the gene transfected with llinc000522 alone.@*Conclusion@#In gastric cancer cells, llinc00052 can affect the invasion and metastasis of gastric cancer by regulating the level of miRNA and affecting the Wnt/β-catenin pathway.
ABSTRACT
Objective@#To investigate the clinical features, diagnosis, occurrence sequence and clonal origin of chronic lymphocytic leukemia complicated with multiple myeloma.@*Methods@#The diagnosis and treatment of one patient with multiple myeloma and chronic lymphocytic leukemia who was admitted to the First Hospital of Jilin University in May 2018 was retrospectively analyzed, and the related literatures were reviewed.@*Results@#This patient began with lumbosacral pain, and he was diagnosed as chronic lymphocytic leukemia complicated with multiple myeloma after bone marrow aspiration, flow cytometry, and blood and urine immunofixation electrophoresis. It is recommended that Rd (lenalidomide + dexamethasone) or MPV (melphalan + prednisone + bortezomib) regimen, but the patient did not receive chemotherapy and died of infectious diarrhea 1 month later.@*Conclusions@#The occurrence of multiple myeloma and chronic lymphoblastic leukemia may originate from the same clone or different new clone. It is very rare that multiple myeloma and chronic lymphoblastic leukemia can co-occur. Therapeutic options tend to be more aggressive multiple myeloma-based regimen.
ABSTRACT
Objective To explore the mechanism of llinc00052 regulating Wnt pathway affecting invasion and metastasis of gastric cancer.Methods SGC-7901 cells were selected from gastric cancer cell lines ,and siRNAs related to INC0005 and had invasion and metastasis effects in gastric cancer cells were screened by binding of IncRNA expression profiles to qRT -PCR.Ilinc00052 and miRNA expression changes were studied by in vitro cell transfection experiments.Through molecular experiments ,the expression of llinc00052 and the effect on Wnt／β-catenin expression were investigated to explore whether llinc 00052 could affect the invasion and metastasis of gastric cancer cells by regulating miRNAs affecting Wnt／β-catenin signaling pathway.Results Transwell chamber test showed that the number of transmembrane cells in the untransfected plasmid group was (134.10 ±4.29),and the number of transmembrane cells in the overexpressed llinc 000522 plasmid group was (169.24 ±6.99)(t＝8.956,P＝0.001). The scratch test showed that the migration distance in the llinc 000522 overexpression transfection plasmid group was significantly higher than that in the no -load plasmid transfection group (r＝0.907,P＜0.01).The clone formation rate of llinc000522 overexpressed transfected plasmid group was significantly higher than that of the empty plasmid group[(92.75 ±6.32)% vs.(73.34 ±9.14)%] (t＝5.998,P＜0.05).Compared with the transfection of blank plasmid,the expressions of Wnt1,Wnt3a,Wnt2 and β-catenin mRNA in the llinc000522 overexpression transfection group were significantly up -regulated(P＜0.05),while the miRNA transfection group had no significant effect on the expression.The expressions of Wnt1,Wnt3a,Wnt2,and β-catenin mRNA were significantly increased [(1.82 ± 0.11),(1.52 ±0.15),(1.42 ±0.21),(1.71 ±0.19)] ( P＜0.05),but their expressions were still lower than those of the genes transfected with llinc000522 alone.Compared with the blank plasmid transfection ,the expressions of Wnt1,Wnt3a,Wnt2 and β-catenin protein in the llinc000522 overexpression transfection group were significantly up-regulated(all P＜0.05),while the miRNA transfection group had no significant effect on its expression.The protein expressions of Wnt1,Wnt3a,Wnt2 and β-catenin were also significantly increased [(1.53 ±0.09),(1.4 ±0.21), (1.33 ±0.07),(1.47 ±0.19)](P＜0.05),but their expressions were still lower than those of the gene transfected with llinc000522 alone.Conclusion In gastric cancer cells, llinc00052 can affect the invasion and metastasis of gastric cancer by regulating the level of miRNA and affecting the Wnt ／β-catenin pathway.
ABSTRACT
Abstract Heterogeneous cell populations of osteo/cementoblastic (O/C) or fibroblastic phenotypes constitute the periodontal dental ligament (PDL). A better understanding of these PDL cell subpopulations is essential to propose regenerative approaches based on a sound biological rationale. Objective Our study aimed to clarify the differential transcriptome profile of PDL cells poised to differentiate into the O/C cell lineage. Methodology To characterize periodontal-derived cells with distinct differentiation capacities, single-cell-derived clones were isolated from adult human PDL progenitor cells and their potential to differentiate into osteo/cementoblastic (O/C) phenotype (C-O clones) or fibroblastic phenotype (C-F clones) was assessed in vitro. The transcriptome profile of the clonal cell lines in standard medium cultivation was evaluated using next-generation sequencing technology (RNA-seq). Over 230 differentially expressed genes (DEG) were identified, in which C-O clones showed a higher number of upregulated genes (193) and 42 downregulated genes. Results The upregulated genes were associated with the Cadherin and Wnt signaling pathways as well as annotated biological processes, including "anatomical structure development" and "cell adhesion." Both transcriptome and RT-qPCR showed up-regulation of WNT2, WNT16, and WIF1 in C-O clones. Conclusions This comprehensive transcriptomic assessment of human PDL progenitor cells revealed that expression of transcripts related to the biological process "anatomical structure development," Cadherin signaling, and Wnt signaling can identify PDL cells with a higher potential to commit to the O/C phenotype. A better understanding of these pathways and their function in O/C differentiation will help to improve protocols for periodontal regenerative therapies.
Subject(s)
Humans , Adult , Osteoblasts/cytology , Periodontal Ligament/surgery , Dental Cementum/cytology , Cadherins/metabolism , Cell Differentiation , Cells, Cultured , Clone Cells , TranscriptomeABSTRACT
Objective To evaluation the method of rapid detection of Methicillin-resistant Stphylococcus aureus (MRSA) ST239 clones with multiplex PCR assay and investigation of the epidemic status of MRSA blood stream infections in Hefei area.Methods Antibiotic susceptibility testing were applied to MRSA isolates from bloodstream infection,rapid screening and confirmation of MRSA ST239 clones by using multiplex PCR,Multilocus Sequence typing (MLST) and Staphyloccoccal Cassette Chromosome mec(SCCmec) typing.Results 51 of 106 clinic isolates Staphylococcus aureus were identified as MRSA,accounting for 48.1％.The resistance rate of MRSA to erythromycin,aminoglycosides and quinolone were significantly higher than Methicillin Sensitive Staphylococcus aureus (MSSA).Both MRSA and MSSA had a high sensitivity to cotrimoxazole,the sensivity rates were 86.3％ and 94.5％,respectively; 47 of 51 isolates of MRSA that detected by MRSA ST239 rapid screening were ST239 clones.Randomly selected 20 positive screening stains were confirmed as MRSA-ST239-SCCmec Ⅲ by MLST and SCCmectyping.Conclusions In Hefei area,nearly half of MRSA bloodstream infections in clinical isolates are MRSA-ST239-SCCmeclⅢ type and serious multidrug-resistance.The rapid detection of ST239 clones by multiplex PCR is a reliable and effective method for large-scale screening in laboratory.
ABSTRACT
BACKGROUND:Transplantation of exogenous stem cells for functional cardiac celldeath or apoptosis easily induced complications after transplantation. Therefore, cardiac progenitor cells of heart itself have been ideal seed cells. OBJECTIVE:To establish stable celllines of cardiac progenitor cells from mouse heart, and to provide ideal cellmodels for studying proliferation and differentiation factors affecting cardiac progenitor cells during adult myocardial cells were damaged. METHODS:(1) Myocardiocytes were isolated from embryonic 15.5 days mice. (2) The cultured myocardiocytes were immortalized using retrovirus SSR69. Immortalized monoclonal myocardial cells were obtained using antibiotic selection and infinite dilution. (3) Monoclonal cellline with the property of cardiac progenitor cells was screened out. RESULTS AND CONCLUSION:(1) Myocardiocytes were successful y isolated and cultured. Partial cells showed obvious beating. (2) Myocardiocytes infected with retrovirus SSR69 were cloned and got 76 clones, then were named as CP15-#, (3) Screening the first 20 clones, the reasonable clones with the characteristics of cardiac progenitor cells were obtained according to myocardial cellmarker genes. The results suggested that immortalized cardiac progenitor cells were established mediated by reversible SV40 T antigen.
ABSTRACT
Objective To study the role of gene Scanning in the clonality analysis of Ig/TCR gene remrangement in children with newly diagnosed acute lymphoblastic leukemia (ALL),and the relationship between the clinical characteristics of ALL and the type of Ig/TCR gene rearrangement.Methods The research was the clinical experimental study.Selected 86 cases of children with ALL who were diagnosed and treated in Department of Hematology-oncology of Guangzhou Women and Children's Medical Center,and All cases were confirmed by bone marrow cell morphology and flow cytometric immunophenotyping.Used multiplex PCR to detecte Ig/TCR gene rearrangements in children with newly diagnosed ALL.Applied gene scanning to analyze the clonality of Ig/TCR gene rearrangement.Results There were 83 cases detected 1 or more than 1 types of Ig/TCR gene rearrangements in 86 cases (96.5％),with 2.52 types each case.There were 56 cases detected at least one monoclonal Ig/TCR gene rearrangement in 61 cases analyzed by gene scanning (91.8％).The detectable rate for lgH,Igκ,TCRγ and TCRδ were 27.91％,27.91％,19.77％ and 24.42％ respectively in 172 of Ig/TCR gene rearrangement.Monoclonal,oligoclonal and polyclonal composition was 58.1％,30.8％ and 11.1％ respectively,the monoclonal as the main component.There was no significant difference between the types and clonality of Ig/TCR gene rearrangement and the clinical characteristics of children with newly diagnosed ALL (P ＞ 0.05).Conclusions Gene scanning can analyze clonality of the Ig/TCR gene rearrangement conveniently and rapidly,thus,it can be possible to select the stable targets for quantitative detection of minimal residual disease minimal residual disease.There is no significant difference between the types and clonality of Ig/TCR gene rearrangement and clinical characteristics of children with newly diagnosed.
ABSTRACT
ObjectiveTo study transcriptional characteristics of type Ⅲ procollagen gene in systemic scleroderma (SS)-derived fibroblast clones and their regulation by Radix Salviae miltiorrhizae(RSM).Methods Eight fibroblast clones with different collagen-producing capacity were previously obtained from patients with SS and normal human controls.Recombinant plasmids containing different deletions of the human alpha 1 chain of type 3 procollagen(COL3A1) gene promoter were constructed,and transiently transfected into the fibroblast clones.Dual-luciferase reporter assay system was used to evaluate the activities of these recombinants in the fibroblast clones and to select a proximal transcriptional regulatory sequence.Then,the fibroblast clones were transfected with the plasmid containing the selected regulatory sequence(phCOLH30.1) followed by the treatment with RSM injection(1 g/L) and active monomers of RSM,including salvianolic acid B(5 mg/L),tanshinone Ⅱ A (5 mg/L),danshensu(20 mg/L) and protocatechuic aldehyde(5 mg/L),for 48 hours.The transfected fibroblast clones receiving no drug treatment served as the water-soluble control,and those treated with only dimethyl sulfoxide as the lipid-soluble control.Subsequently,the fibroblasts were lysed and subjected to the quantification of cellular proteins and determination of luciferase activity.The activity of recombinant promoters was compared by t test for the selection of proximal transcriptional regulatory sequence,and the activity of phCOLH30.1 by two-way analysis of variance in the RSM-interfering test(if there was interaction,one-way analysis of variance was conducted; and if there was no interaction,the main effect was tested after the removal of interaction item).ResultsOf the 6 recombinants,the recombinant containing COL3A1 proximal promoter from -96 bp to +16 bp(phCOLH30.1) showed the highest transcriptional activity in nearly all of the fibroblast clones,and the activity was positively correlated with the collagen-producing capacity of fibroblast clones.Compared with the water-soluble control,RSM injection significantly downregulated the activity of phCOLH30.1 in fibroblast clones with high and low collagen-producing capacity from patients with SS (2.261 ± 0.619 vs.3.879 ± 0.309,1.462 ± 0.291 vs.2.150 ± 0.262,both P ＜ 0.01) and normal human controls (1.681 ± 0.263 vs.3.039 ± 0.271,1.121 ± 0.361 vs.2.223 ± 0.247,both P ＜ 0.01),salvianolic acid B decreased the phCOLH30.1 activity in SS-derived high collagen-producing fibroblast clones (2.309 ± 0.524,P ＜ 0.01 ) and in the normal control fibroblast clones with high and low collagen-producing capacity (2.126 ± 0.320 and 1.976 ± 0.362,both P ＜ 0.05).Tanshinone Ⅱ A only downregulated the phCOLH30.1 activity in SS-derived high collagen-producing fibroblast clones compared with the lipid-soluble control(2.975 ± 0.666 vs 5.379 ± 0.238,P ＜ 0.01 ).Neither danshensu nor protocatechuic aldehyde showed inhibitory effects on phCOLH30.1 activity in SS-derived or normal control fibroblast clones.ConclusionsThe type Ⅲ procollagen gene is activated at the transcriptional level in high collagen-producing fibroblast clones from patients with SS,and the activation could be suppressed by RSM injection,salvianolic acid B and tanshinone Ⅱ A.
ABSTRACT
Objective To investigate the effect of dysfunction of T lymphocytes on clonal haematogenesis in patients with myelodysplastic syndrome (MDS). Methods The cytogentics, the subsets of lymphocytes and their activation in 76 patients with MDS were analyzed. Results There were 36 patients with normal karyotype and 40 patients with abnormal karyotype. The incidence of abnormal karyotype were 52.6 %. There were 24 cases (60.0 %) with trisomy 8 (+8) in 40 cases of abnormal karyotype. The expression rates of CD+3 CD-19 cells, CD+3 CD-4 CD+8 cells and CD+3 HLA-DR+ cells in MDS were significantly increased, and CD-3 (CD16CD56)+ cells were significantly lower than that in control group. The expression rates of CD+3 (CD16CD56)+ cells in MDS with abnormal karyotype were significantly higher than that in control group. The expression rates of CD+3 CD+4 CD-8 cells in +8 MDS were significantly lower than that in MDS patients with normal karyotype and with other abnormal karyotype. The ratio of CD4/CD8 in +8 MDS were significantly lower than that in control group. Conclusion The abnormalities of T cell subsets and functions in patients with MDS were observed and the proliferation of malignant clone was prevalent which indicated a poor prognosis in MDS with abnormal karyotype. Dysfunction of immunosurveillance was more aggravated in +8 MDS, which led to excess proliferation of malignant clone and over inhibition of remaining haematogenesis.
ABSTRACT
Objective To study the transcriptional regulation of type Ⅰ procollagen gene in systemic scleroderma(SS)-derived high collagen-producing fibroblast clones by Radix Salviae Miltiorrhizae(RSM).Methods Fibroblast clones with different collagen-producing capacity were previously obtained from patients with SS and normal human controls,and divided into 5 groups to be treated with RSM(1 g/L)injection,its water-soluble active monomers including sodium danshensu(20 mg/L),salvianolic acid B(5 mg/L)and protocatechuic aldehyde(5 mg/L),and lipid-soluble active monomer(tanshinone Ⅱ A,5mg/L)respectively.The fibroblast clones incubated with no drugs served as the water soluble negative control group,and those with dimethyl sulfoxide(DMSO)as the lipid soluble negative control group.MTT assay was performed to evaluate the proliferation of the fibroblast clones after 1-,3-,5-,and 7-day treatment,transient transfection and dualluciferase reporter assay system to quantify the relative activity of collagen type Ⅰ,alpha 1(COL1A1)proximal promoter in these fibroblast clones.Results The inhibitory effect of RSM and its active monomers on the proliferation of fibroblast clones was inapparent within the initial 3 days(P ＞ 0.05),but was enhanced with incubation time.A significant difference was observed in the proliferation level of fibroblast clones between RSM group and water-soluble negative control group on day 5(q′ =3.22,P ＜ 0.01),between RSM,salvianolic acid B,protocatechuic aldehyde groups and the water-soluble negative control group(q′ =4.74,3.03,2.56,all P ＜0.05)on day 7,and between tanshinone Ⅱ A and lipid-soluble negative control group on day 5 and 7(t =2.22,2.15,both P ＜ 0.05).RSM injection,tanshinone Ⅱ A and protocatechuic aldehyde significantly inhibited COL1A1 proximal promoter activity in SS-derived and normal control fibroblast clones(all P ＜ 0.01),and the former two drugs preferentially downregulated COL1A1 proximal promoter activity in SS-derived high collagenproducing fibroblast clones.Significantly different COL1A1 proximal promoter activity was observed in SS-derived high and low collagen-producing fibroblast clones between water-soluble negative control group and RSM injection group(12.019 ± 0.830 vs.4.445 ± 1.061,5.388 ± 0.480 vs.2.856 ± 0.597,F=31.78,P＜ 0.01),and between lipid-soluable negative control group and tanshinone Ⅱ A group(14.155 ± 0.672 vs.9.638 ±0.854,4.299 ± 0.252 vs.3.192 ± 0.450,F=24.10,P＜ 0.01).Conclusions RSM inhibits the transcription of COL1A1 gene in SS-derived high collagen-producing fibroblast clones,which may be mainly attributed to tanshinone Ⅱ A and protocatechuic aldehyde.
ABSTRACT
BACKGROUND/AIMS: Idiopathic pulmonary fibrosis (IPF) is defined pathologically by usual interstitial pneumonia (UIP), and contains characteristic discrete areas of fibroblasts, myofibroblasts, and newly formed collagen, termed "fibroblast foci". A new hypothesis postulates that IPF results from epithelial injury and abnormal wound repair without preceding chronic inflammation. We explored the hypothesis that fibroblasts in the fibroblast foci of IPF undergo neoplastic monoclonal proliferation rather than reactive polyclonal proliferation. METHODS: We obtained fibroblasts from 24 fibroblast foci in seven female patients with IPF, endothelial cells from ten plexiform lesions of two female patients with idiopathic pulmonary arterial hypertension (IPAH) as a positive control for monoclonality, and lung parenchymal cells by microdissection of each formalin-fixed paraffin-embedded block of lung. To analyze clonality, we performed the human androgen-receptor gene methylation assay (HUMARA). DNA released by protein K digestion was subjected to polymerase chain reaction (PCR) amplification with prior digestion with and without the methylation-sensitive restriction enzyme HhaI. Then, we calculated the clonality ratio after electrophoretic analysis of the PCR amplification product. A clonality ratio 0.25, suggesting polyclonality, whereas 7 of 10 plexiform lesions had clonality ratios <0.25 suggesting monoclonality. CONCLUSIONS: The polyclonality of the fibroblast foci in IPF suggests that the fibroblast proliferation in IPF is not neoplastic, but is reactive in nature
Subject(s)
Female , Humans , Bacterial Outer Membrane Proteins , Clone Cells , Collagen , Digestion , DNA , Endothelial Cells , Fibroblasts , Hypertension , Hypertension, Pulmonary , Idiopathic Pulmonary Fibrosis , Inflammation , Lung , Methylation , Microdissection , Myofibroblasts , Polymerase Chain Reaction , X ChromosomeABSTRACT
Objective To compare the cytokine expression profile of 3 CD8+, 3 CD4+ and 3 gamma delta-positive T cell clones derived from the synovial fluids of reactive arthritis (ReA) patients and to explore the immunological pathogenesis of ReA. Methods Complementary DNA-based microarrays containing genes for 56 cytokines were used for screening the expression profile of 3 CD8+, 3 CD4+ and 3 gamma delta-positive T cell clones derived from the synovial fluids of 3 reactive arthritis (ReA) patients. Results Transcripts encoding for interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha were expressed among all CD8+ and CD4+ T cell clones by microarray. Expression of these cytokines could be verified by RT-PCR in 14 out of 15 microarray experiments. Lymphotoxin (LT)-alpha and granulocyte macrophage colony-stimulating factor (GM-CSF) were also consistently expressed among CD4+ cells. However, ??+T cells revealed a different cytokine pattern, mainly expressing transforming growth factor (TGF)-beta 2 and GM-CSF. Conclusion CD8+ and CD4+ T cells mainly reveale a Th1-mediated profile, whereas ??+T cells expresse less pro-inflammatory cytokines resembling a Th3-driven pattern. T lymphocyte clones from the joint of ReA patients exhibit different cytokine expression profiles refelecting their different roles in pathogenesis.
ABSTRACT
BACKGROUND: Tumors are usually considered to be clonal progeny of single transformed cells. Carcinosarcomas and malignant mixed epithelial tumors are examples where controversies exist regarding the singularity or multiplicity of their cell of origin. METHODS: The authors examined the clonality of carcinosarcomas (7 cases) and malignant mixed epithelial tumor (5 cases) in female patients by X-chromosome inactivation as a marker. Each component of the tumors were picked up by the laser capture microscope. The polymorphic exon 1 CAG trinucleotide repeat in the X-linked human androgen receptor (HUMARA) gene was amplified by a polymerase chain reaction before and after treatment of the methylation-sensitive endonuclease HpaII. RESULTS: Eleven cases were informative for clonality determination. Six out of seven carcinosarcomas and three out of four malignant mixed epithelial tumors revealed the same patterns of X-chromosome inactivation, which suggests that they are monoclonal. In contrast, the patterns of X-chromosome inactivation were different between the two tumor components in each cases of carcinosarcoma and malignant mixed epithelial tumor, indicating that they are of polyclonal origin. CONCLUSIONS: These observations show that although most of carcinosarcomas and malignant mixed epithelial tumors are of monoclonal origin, some of them are of polyclonal origin. This finding suggests that these tumors are genuinely polyclonal, and that they originated in the neoplastic transformation of more than one somatic cells
Subject(s)
Female , Humans , Carcinosarcoma , Deoxyribonuclease HpaII , Exons , Polymerase Chain Reaction , Receptors, Androgen , Trinucleotide RepeatsABSTRACT
A Review] The biological characteristics of viral L6565 leukemia cell clone were as follows: (1) The chromosome counts varied 38～114 , and stem cells were 42; (2) Virus particles type A and type C found in the cytoplasm of clone cells; (3) X-C assays were positive, c- myc and c- fos gene overexpressed in clone cells; (4) Differential markers CD4, CD8, CD45R were negative, CD45RO ? were positive; (5) The supernatant of clone cells could induce T or B lymphocytic leukemia/lymphoma and granulocytic leukemia in SSB strain mice. The leukemogenic effect of concentrate supernatant was stronger than non-concentrate supernatant( P
ABSTRACT
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.
ABSTRACT
Object To explore, on the whole, the anticancer activity and mechanism of the aqueous extract of Ruixiang Langdu (Stellera chamaejasme L.) (SCLA). Methods Mice were given different i.g. doses of SCLA and their serum collected at different intervals after drug administration. The effects of the serum on the proliferation of mouse L 1210 leukemic cells were observed with MTT assay, clone formation and incorporation of [ 3H] TdR into DNA of the cells. Results Serum SCLA collected 1, 2, 4, 8 h after administration of 3, 6, and 12 g/kg SCLA showed significant decrease of MTT formazans and clone formation. Serum SCLA collected 2 h after drug administration showed more strong inhibition of cancer cell proliferation. It also inhibited the incorporation of [ 3H] TdR into DNA of L 1210 cells. Conclusion SCLA showed a remarkable inhibitory effect on proliferation and DNA synthesis of cancer cells cultured in vitro, which may be the main anticancer mechanisms of SCLA.
ABSTRACT
Human LAK cells induced from peripheral blood mononuclear cells with recombinant interleukin2 (rIL-2) incubation, can be cloned in semisolid medium and proliferated well in liquid cultures containing rIL-2. Phynotypie analysis by flow cytometry showed that these cloned LAK cells were T_3(+), T_4(-), T_8(+), Tac(+), DR(+) and NKH1(-)/(+). Cultured with 1?10~5/ml normal bone marrow mononuelear cells (BMMNC)for 0.5-hr prior to semisolid culture system for CFU-GM, the cloned LAK ceils at 2?10~5/ml showed inhibitory effect on CFU-GM that the colonies were decreased by 43.8% of control though 0.5 or 1?10~5/ml LAK cells did not. Morever, a dose-dependent inhibition of CFU-GM was noted when LAK cells pre-inbated with BMMNC for 4-hr in doses of 0.5 -4?10~5/ml that the inhibitory rates of CFU-GM were 58.8, 67.5, 88.8 and 96.3% at0.5, 1, 2, 4?10~5/ml LAK cells addition respectively. Using double-layer agar cultures, inhibitory activity for CFU-GM was also observed only when LAK cells at 4?10~5/ml without contacted with BMMNC. No significant difference of inhibitory effects on CFUGM growth was found between autologous and allogenie clotted LAK cells under same condition. These data suggeste that LAK cells inhibited growth of myeloid progenitor cells without MHC-restriction and the mechanism might involve cell-to-cell interaction and/or some soluble factors secreted by LAK cells.
ABSTRACT
A clone system designated SAC was established from a SRS-82 mouse ascitic turnout cell line by using limited dilution method. Clones SAC-ⅡB_2, SAC-ⅡB_3 of this system were found to be different in their morphology, growth pattern, karyotypes and tumorigenicity from their parent cell line. Both the two clones and their parent cell line were X-C assay positive, and type A and type C virus particles were found in the cytoplasm and cell surface of these cells electronmicroscopically. Cell surface markers(ALS, Thy, SmIg G) of these two clones and their parent cell line were investigated by ABC immunohistochemical method and immunofluorescent staining technique, the results of which indicate that all of them belong to T cell origin.