[Cloning of human KIAA1173 gene and biological characterization of transfected 6-10B cells].
OBJECTIVE
To establish a 6-10B cell line with stable expression of KIAA1173 gene and study the biological behaviors of the cells.


METHODS
The total RNA was extracted from normal skeletal muscular tissues for cloning of KIAA1173 gene by means of RT-PCR which was subsequently introduced into pcDNA3.1 (+) vector. The recombinant eukaryotic expression vector pcDNA 3.1(+)-KIAA1173 was constructed and identified by endonuclease digestion and sequencing before transfection into 6-10B cells via lipofectamine with the empty vector as the control. The positive cell clones were obtained by G418 selection. Stable expression of KIAA1173 gene in the transfected 6-10B cells was determined by RT-PCR, in situ hybridization and immunocytochemistry, and the biological behaviors of the transfected cells were observed by MTT assay, cell invasion assay and tumorigenesis assay in nude mice.


RESULTS
High expression of KIAA1173 at both mRNA and protein levels was observed in the transfected 6-10B cells. The capability of proliferation, invasion and tumorgenicity of the KIAA1173-transfected cells in nude mice was lowered in comparison with those of the cells transfected with pcDNA3.1 (+) vector (P<0.05).


CONCLUSIONS
KIAA1173 genes may function as a potential tumor suppressor of nasopharyngeal carcinoma both in vitro and in vivo. The 6-10B cell line expressing KIAA1173 has been obtained, which can be helpful for further study of KIAA1173 gene.
Zhang
San-quan
SQ
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. zhangsq2000@sohu.com
Peng
Hong
H
Jiang
Hui-yong
HY
Hu
Hai
H
Zhang
Jin-hua
JH
Yao
Kai-tai
KT
Zhao
Tong
T
D000818:Animals; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D005057:Eukaryotic Cells / Q000378:metabolism; D016147:Genes, Tumor Suppressor; D005822:Genetic Vectors; D006801:Humans; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D008819:Mice, Nude; D009303:Nasopharyngeal Neoplasms / Q000235:genetics* / Q000473:pathology; D009368:Neoplasm Transplantation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis / Q000235:genetics
C432970:HHATL protein, human; D008565:Membrane Proteins; D011994:Recombinant Proteins; D025521:Tumor Suppressor Proteins