[Construction of human mucosa oral epithelial cell lines overexpressing telomerase reverse transcriptase gene mediated by lentivirus].
OBJECTIVE
To construct a cell line of oral mucosa epithelial cells that stably express human telomerase reverse transcriptase (hTERT) by lentiviral vectors, approaches for the establishment of stable and efficient immortalized oral mucosa epithelial cell lines were explored.


METHODS
Whole RNA was extracted from 293T cells. The hTERT gene was amplified by polymerase chain reaction (PCR) and cloned into the lentiviral vector as pLVX-puro-hTERT. The lentivirus particles were successfully packaged and used to infect primary oral epithelial cells. The positive cell clones were selected by puromycin. Finally, the expression of hTERT was examined by real-time fluorescent quantitative PCR (qRT-PCR) and Western blot analysis.


RESULTS
The sequencing results confirmed the construction of the recombinant lentivirus pLVX-puro-hTERT. The morphology of infected cells was similar to that of normal oral mucosal epithelial cells, with a cobble stone-like appearance. The qRT-PCR and Western blot results showed that hTERT was overexpressed in infected cells compared with the normal group (P<0.05).


CONCLUSIONS
The oral epithelial cell line with stable expression of hTERT was successfully established by the lentivirus, which provides an experimental basis for the establishment of a highly efficient and stable oral epithelial immortalized cell line.
School of Stomatology, Lanzhou University, Lanzhou 730000, China.
Zeng
Z
Sa
Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730000, China.
Qin
Q
Xiaodong
School of Stomatology, Lanzhou University, Lanzhou 730000, China.
He
H
Xiangyi
School of Stomatology, Lanzhou University, Lanzhou 730000, China.
Che
C
Chunxiao
School of Stomatology, Lanzhou University, Lanzhou 730000, China.
Zhang
Z
Xiao
School of Stomatology, Lanzhou University, Lanzhou 730000, China.
Xie
X
Siyu
School of Stomatology, Lanzhou University, Lanzhou 730000, China.
Sun
S
Guijun
School of Stomatology, Lanzhou University, Lanzhou 730000, China.
Wang
W
Lihe
D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus; D009055:Mouth; D009092:Mucous Membrane; D060888:Real-Time Polymerase Chain Reaction; D019098:Telomerase
C509186:TERT protein, human; D019098:Telomerase
human telomerase reverse transcriptase; immortalization; lentivirus; oral mucosa epithelial cells; stable expression