Enhancement of cell proliferation in various mammalian cell lines by gene insertion of a cyclin-dependent kinase homolog
| Autor: | Michael J. Betenbaugh, Pratik Jaluria, Joseph Shiloach, Konstantinos Konstantopoulos |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: |
Cell Survival
lcsh:Biotechnology Genomics Cell Count CHO Cells Biology Protein Serine-Threonine Kinases Electron Transport Complex IV 03 medical and health sciences 0302 clinical medicine Cricetulus Cyclin-dependent kinase lcsh:TP248.13-248.65 Cricetinae Cell Adhesion Animals Humans Insertion Gene Interphase 030304 developmental biology Cell Proliferation Oligonucleotide Array Sequence Analysis 0303 health sciences Cell growth Contact Inhibition Gene Expression Profiling Cyclin-Dependent Kinases Cell biology Up-Regulation Gene expression profiling Phenotype Cell culture 030220 oncology & carcinogenesis biology.protein Female DNA microarray Biotechnology Research Article HeLa Cells |
| Zdroj: | BMC Biotechnology BMC Biotechnology, Vol 7, Iss 1, p 71 (2007) |
| ISSN: | 1472-6750 |
| Popis: | BackgroundGenomics tools, particularly DNA microarrays, have found application in a number of areas including gene discovery and disease characterization. Despite the vast utility of these tools, little work has been done to explore the basis of distinct cellular properties, especially those important to biotechnology such as growth. And so, with the intent of engineering cell lines by manipulating the expression of these genes, anchorage-independent and anchorage-dependent HeLa cells, displaying markedly different growth characteristics, were analyzed using DNA microarrays.ResultsTwo genes, cyclin-dependent kinase like 3 (cdkl3) and cytochrome c oxidase subunit (cox15), were up-regulated in the faster growing, anchorage-independent (suspension) HeLa cells relative to the slower growing, anchorage-dependent (attached) HeLa cells. Enhanced expression of either gene in the attached HeLa cells resulted in elevated cell proliferation, though insertion ofcdkl3had a greater impact than that ofcox15. Moreover, flow cytometric analysis indicated that cells with an insert ofcdkl3were able to transition from the G0/G1 phases to the S phase faster than control cells. In turn, expression ofcox15was seen to increase the maximum viable cell numbers achieved relative to the control, and to a greater extent thancdkl3. Quantitatively similar results were obtained with two Human Embryonic Kidney-293 (HEK-293) cell lines and a Chinese Hamster Ovary (CHO) cell line. Additionally, HEK-293 cells secreting adipocyte complement-related protein of 30 kDa (acrp30) exhibited a slight increase in specific protein production and higher total protein production in response to the insertion of eithercdkl3orcox15.ConclusionThese results are consistent with previous studies on the functionalities ofcdkl3andcox15. For instance, the effect ofcdkl3on cell growth is consistent with its homology to thecdk3gene which is involved in G1 to S phase transition. Likewise, the increase in cell viability due tocox15expression is consistent with its role in oxidative phosphorylation as an assembly factor for cytochrome c oxidase and its involvement removing apoptosis-inducing oxygen radicals. Collectively, the present study illustrates the potential of using microarray technology to identify genes influential to specific cellular processes with the possibility of engineering cell lines as desired to meet production needs. |
| Databáze: | OpenAIRE |
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