[Analyzed the molecular interaction network of tumor suppressor gene 14-3-3 sigma in lung cancer cell based on stable isotope labeling by amino acids in cell culture technology].
OBJECTIVE
To analysis the molecular interaction network of 14-3-3 sigma in non small cell lung cancer (NSCLC) cells.


METHODS
Established stable over-expressed 14-3-3 sigma protein PG cells, MTT assay was used to assess the growth rate of PG cells. Though stable isotope labeling by amino acids in cell culture (SILAC) and Mass spectrometry (MS) technology, to identify difference expressed proteins caused by over expressed 14-3-3 sigma. The protein expressed >2 or <0.5 times was termed as the differential protein. By searching Human protein reference database (HPRD) and Kyoto encyclopedia of genes and genomes (KEGG), established the molecular interaction network of tumor suppressor gene 14-3-3 sigma.


RESULTS
The growth rate of over-expressed 14-3-3 sigma PG cell was obviously slower down compared to vector PG cells. A database including 147 differential protein was established. And a molecular interaction network of 14-3-3 sigma containing 26 protein was constructed.In this network, the expression of CSNK2A1 (casein kinase II subunit alpha), involved in numerous cellular processes, such as cell cycle progression, apoptosis and transcription, was the most significantly increased. A DNA repair protein, MEN1 (Menin) which functions as a transcriptional regulator was the most significantly decreased.


CONCLUSIONS
After stable transfected with 14-3-3 sigma gene, growth rate of PG cells was inhibited, the proteins associated with cell cycle, DNA damage repair mechanisms were significantly changed, and constructed the molecular interaction network.
State Key Laboratory of Molecular Oncology, Cancer Institute (Hospital), Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing 100021, China.
Ting
T
Xiao
Wei
W
Mi
Min
M
Li
Bang-rong
BR
Cao
Lin
L
Feng
Shu-jun
SJ
Cheng
Yan-ning
YN
Gao
D048948:14-3-3 Proteins / Q000235:genetics*; D000596:Amino Acids; D014408:Biomarkers, Tumor / Q000235:genetics*; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005095:Exoribonucleases / Q000235:genetics*; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D008175:Lung Neoplasms / Q000235:genetics*; D013058:Mass Spectrometry; D014162:Transfection
D048948:14-3-3 Proteins; D000596:Amino Acids; D014408:Biomarkers, Tumor; D005095:Exoribonucleases; C413442:SFN protein, human