ABSTRACT
OBJECTIVE@#To explore the inhibitory role of spermatogenesis-associated gene 12 (SPATA12) on tumor cell proliferation and its possible mechanism.@*METHODS@#The expression pattern of SPATA12 in testicular tumors was investigated by in situ hybridization analysis using tissue microarrays. The effects of SPATA12 on tumor cell proliferation and colony formation was detected by 3-(4.5-dimethylthiazol-2- yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and colonyforming assays, respectively. The changes of expression level of cell cycle genes in tumor cells were detected by reverse transcription polymerase chain reaction(RTPCR).@*RESULTS@#In situ hybridization analysis showed that the SPATA12 was highly expressed in normal adult testis, but lacking in testicular tumors such as seminoma. MTT assay and colony-forming assay indicated that the exogenous expression of SPATA12 could suppress both tumor cell proliferation and colony formation. RT-PCR showed that the expression of cyclin A1 gene was markedly suppressed and the level of cyclin D1 was somewhat reduced following SPATA12 transfection. However, no obvious changes were observed in mRNA expression of cyclin B1 or cyclin E1 after SPATA12 transfection.@*CONCLUSION@#SPATA12 could be an inhibitor during the development of tumor via regulation of cell cycle genes.
Subject(s)
Humans , Male , Cell Line, Tumor , Cell Proliferation , Genes, Tumor Suppressor , Genes, cdc , HeLa Cells , Homeodomain Proteins , Genetics , Metabolism , Testicular Neoplasms , Genetics , Pathology , TransfectionABSTRACT
In this study, DNAzymes against cyclin D1 (cyclin D1-DRz) were designed according to the secondary structure of cyclin D1 mRNA which was computed with RNAdraw and Mfold. Cyclin D1-DRz were transfected into tumor cell line u251 and HeLa by oligofectamine. The expression of cyclin D1 was detected by RT-PCR. It was shown that the expression of cyclin D1 gene was suppressed obviously, and the expressions of other cell-cycle related genes such as cyclin E1, cyclin A1 and cyclin B1 were also declined. The cell cycle analysis of tumor cells tansfected with cyclin D1-DRz revealed an arrestment in the G0/G1 phase. In conclusion, the approach is effective and feasible for designing DNAzyme. Cyclin D1-DRz is useful for interfering with the cell cycle procession of tumor cells.
Subject(s)
Humans , Base Sequence , Cell Cycle , Cyclin D1 , Genetics , Metabolism , DNA, Catalytic , Genetics , Metabolism , Pharmacology , HeLa Cells , Molecular Sequence Data , RNA, Messenger , Genetics , Metabolism , TransfectionABSTRACT
Objective To study the genetic diversity of germplasm resources for Potentilla discolor.Methods Twelve species of germplasm resources for P.discolor were analyzed by ISSR molecular markers.To make up the systematic diagram of genetic relationship by POPGENE 1.32 software,cluster by UPGMA method,and establish the dendrogram.Results A total of 128 ISSR bands were scored for 12 primers,among which 101 were polymorphic bands.The average percentage of polymorphic bands was 78.91%.Genetic similarity coefficient was changed from 0.179 2 to 0.632 5.By cluster analysis,the geographical distribution is mutually related to the relationship of germplasm resources for P.discolor and it was also showed some of P.discolor from the same region were in the same group which presented the rule of geographical distribution in the tested materials.Conclusion The diversity level of the different germplasm resources for P.discolor higher and the relationship of P.discolor correlates with the geographical location in some way.