Construction and identification of recombinant lentivirus vector for microRNA-223 overexpression and suppression.
- Author:
Yun WANG
;
Ning JI
;
Min ZHOU
;
Lu JIANG
;
Qianming CHEN
- Publication Type:Journal Article
- MeSH: Base Sequence; Cell Line, Tumor; Genetic Vectors; Humans; Lentivirus; MicroRNAs; metabolism; Real-Time Polymerase Chain Reaction
- From: West China Journal of Stomatology 2015;33(5):451-455
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct microRNA-223 overexpression and suppression lentivirus vectors and determine their effects after infecting oral squamous cell carcinoma (OSCC) cell line.
METHODSLentivirus vectors GV229 and GV232 were cut by the restriction sites of Age I and EcoR I and connected to the target gene, which contained mature microRNA-223 and microRNA-223 oligonucleotide. Real-time polymerase chain reaction (PCR) method was used to detect the microRNA-223 expression level after infecting the recombinant lentivirus vector into the OSCC cell line.
RESULTSThe successful construction of microRNA-223 recombinant lentivirus vectors was confirmed by the PCR method and DNA sequencing. HN-30 cell infected with microRNA-223 overexpression vector showed a significant increased in microRNA-223 expression, whereas HN-30 cell infected with microRNA-223 inhibitor vector suppressed microRNA-223 expression.
CONCLUSIONThe microRNA-223 overexpression and suppression lentivirus vectors are successfully constructed. These vectors could alter the expression level of microRNA-223 in OSCC cell line significantly, and provide a stable cell line for functional studies in the future.