To study the role of cyclinD 1 and CDK4 in malignant transformation of human fetal lung diploid fibroblast cell line （2BS） induced by silica. Methods Recombination vectors with sense and antisense pXJ41-cyclinD1 and...To study the role of cyclinD 1 and CDK4 in malignant transformation of human fetal lung diploid fibroblast cell line （2BS） induced by silica. Methods Recombination vectors with sense and antisense pXJ41-cyclinD1 and pXJ41-CDK4 were constructed, and then transfected into the malignant transformed cells induced by silica, respectively. At the same time, pXJ41-neo was used as the control. Results During the progress of the malignant transformation of 2BS cells induced by silica, cyclinD 1 and CDK4 were overexpressed. Antisense RNA suppressed cyclinD 1 and CDK4 gene expression in the antisense pXJ41-cyclinD1 and pXJ41-CDK4 transfected cells. Antisense RNA led to cell cycle arrest, resulting in lengthened G1 phase （the percentages of cells in the G1 phase changed from 45.1% to 52.7% and 58.0% for cyclinD1 and CDK4 transfected cells, respectively）, and eventually attenuated the increase of the proliferation of malignant transformed cells induced by silica. Compared with malignant transformed cells induced by silica, cells transfected with antisense pXJ41-cyclinD1 and pXJ41-CDK4 showed obviously reduced growth rates. On the 8th day, the suppression rates were 58.69 and 77.43% （the growth rate of malignant transformed cells induced by silica was 100%）, doubling time changed from 21.0 h to 31.4 h and 21.0 h to 42.7 h, respectively, the growth capacities on soft agar of cells transfected by antisense pXJ41-cyclinD1 and pXJ41-CDK4 decreased obviously. Conclusion CyclinD 1 and CDK4 play an important role in maintaining transformed phenotype of the cancer cells.展开更多
基金This work was supported by grants from National Natural Science Foundation of China (30028019) and 973 National Key BasicResearch and Development Program (2002 CB 512906).
文摘To study the role of cyclinD 1 and CDK4 in malignant transformation of human fetal lung diploid fibroblast cell line （2BS） induced by silica. Methods Recombination vectors with sense and antisense pXJ41-cyclinD1 and pXJ41-CDK4 were constructed, and then transfected into the malignant transformed cells induced by silica, respectively. At the same time, pXJ41-neo was used as the control. Results During the progress of the malignant transformation of 2BS cells induced by silica, cyclinD 1 and CDK4 were overexpressed. Antisense RNA suppressed cyclinD 1 and CDK4 gene expression in the antisense pXJ41-cyclinD1 and pXJ41-CDK4 transfected cells. Antisense RNA led to cell cycle arrest, resulting in lengthened G1 phase （the percentages of cells in the G1 phase changed from 45.1% to 52.7% and 58.0% for cyclinD1 and CDK4 transfected cells, respectively）, and eventually attenuated the increase of the proliferation of malignant transformed cells induced by silica. Compared with malignant transformed cells induced by silica, cells transfected with antisense pXJ41-cyclinD1 and pXJ41-CDK4 showed obviously reduced growth rates. On the 8th day, the suppression rates were 58.69 and 77.43% （the growth rate of malignant transformed cells induced by silica was 100%）, doubling time changed from 21.0 h to 31.4 h and 21.0 h to 42.7 h, respectively, the growth capacities on soft agar of cells transfected by antisense pXJ41-cyclinD1 and pXJ41-CDK4 decreased obviously. Conclusion CyclinD 1 and CDK4 play an important role in maintaining transformed phenotype of the cancer cells.
