PMID- 17945021
OWN - NLM
STAT- MEDLINE
DCOM- 20080205
LR  - 20221109
IS  - 1472-6750 (Electronic)
IS  - 1472-6750 (Linking)
VI  - 7
DP  - 2007 Oct 18
TI  - Enhancement of cell proliferation in various mammalian cell lines by gene 
      insertion of a cyclin-dependent kinase homolog.
PG  - 71
AB  - BACKGROUND: Genomics 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. RESULTS: Two 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 of cdkl3 had a greater impact than 
      that of cox15. Moreover, flow cytometric analysis indicated that cells with an 
      insert of cdkl3 were able to transition from the G0/G1 phases to the S phase 
      faster than control cells. In turn, expression of cox15 was seen to increase the 
      maximum viable cell numbers achieved relative to the control, and to a greater 
      extent than cdkl3. 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 either cdkl3 or 
      cox15. CONCLUSION: These results are consistent with previous studies on the 
      functionalities of cdkl3 and cox15. For instance, the effect of cdkl3 on cell 
      growth is consistent with its homology to the cdk3 gene which is involved in G1 
      to S phase transition. Likewise, the increase in cell viability due to cox15 
      expression 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.
FAU - Jaluria, Pratik
AU  - Jaluria P
AD  - National Institute of Diabetes & Digestive & Kidney Diseases, National Institutes 
      of Health, Biotechnology Unit, Building 14A, Room 170, Bethesda, MD 20892, USA. 
      pratikj@mail.nih.gov
FAU - Betenbaugh, Michael
AU  - Betenbaugh M
FAU - Konstantopoulos, Konstantinos
AU  - Konstantopoulos K
FAU - Shiloach, Joseph
AU  - Shiloach J
LA  - eng
GR  - R01 AR053358/AR/NIAMS NIH HHS/United States
GR  - ImNIH/Intramural NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, N.I.H., Intramural
DEP - 20071018
PL  - England
TA  - BMC Biotechnol
JT  - BMC biotechnology
JID - 101088663
RN  - EC 1.9.3.1 (Electron Transport Complex IV)
RN  - EC 2.7.1.- (CDKL3 protein, human)
RN  - EC 2.7.11.1 (Protein Serine-Threonine Kinases)
RN  - EC 2.7.11.22 (Cyclin-Dependent Kinases)
SB  - IM
MH  - Animals
MH  - CHO Cells
MH  - Cell Adhesion/physiology
MH  - Cell Count
MH  - *Cell Proliferation
MH  - Cell Survival/physiology
MH  - Contact Inhibition
MH  - Cricetinae
MH  - Cricetulus
MH  - Cyclin-Dependent Kinases/genetics/*metabolism
MH  - Electron Transport Complex IV/biosynthesis/metabolism
MH  - Female
MH  - Gene Expression Profiling
MH  - HeLa Cells
MH  - Humans
MH  - Interphase/physiology
MH  - Oligonucleotide Array Sequence Analysis/standards
MH  - Phenotype
MH  - Protein Serine-Threonine Kinases/biosynthesis/metabolism
MH  - Up-Regulation
PMC - PMC2164945
EDAT- 2007/10/20 09:00
MHDA- 2008/02/06 09:00
PMCR- 2007/10/18
CRDT- 2007/10/20 09:00
PHST- 2007/06/05 00:00 [received]
PHST- 2007/10/18 00:00 [accepted]
PHST- 2007/10/20 09:00 [pubmed]
PHST- 2008/02/06 09:00 [medline]
PHST- 2007/10/20 09:00 [entrez]
PHST- 2007/10/18 00:00 [pmc-release]
AID - 1472-6750-7-71 [pii]
AID - 10.1186/1472-6750-7-71 [doi]
PST - epublish
SO  - BMC Biotechnol. 2007 Oct 18;7:71. doi: 10.1186/1472-6750-7-71.