Quantitative comparison of MIP-3α mRNA level in different mucosal epithelial cells
- VernacularTitle:不同黏膜上皮细胞中巨噬细胞炎性蛋白-3α转录水平的荧光定量RT-PCR比较分析
- Author:
Tong GAO
;
Chenli QIU
;
Hui ZHAO
;
Qiang LIU
;
Yiming SHAO
;
Guibo YANG
- Publication Type:Journal Article
- Keywords:
Mucosal epithelial cells;
Macrophage inflammatory protein-3α;
Real-time RT-PCR;
Quantitative comparison
- From:
Chinese Journal of Microbiology and Immunology
2008;28(5):458-462
- CountryChina
- Language:Chinese
-
Abstract:
Objective To compare the mRNA level of macrophage inflammatory protein-3α(MIP-3α) in 3 different mucosal epithelial cell lines. Methods RNA standards were prepared by in vitro transcription. One-step real-time RT-PCR was established and optimized using TaqMan EZ RT-PCR Core Reagents with TaqMan probes and primers specific to human MIP-3α mRNA sequence. The specificity of one-step real-time RT-PCR method was confirmed by sequencing the PCR products. The sensitivity and reproducibility of the method was examined by repeating the test 8 times with the same sample. Results The one-step real-time RT-PCR with a wide detection range is sensitive, reproducible. It was found that MIP-3α mRNA level in Caco-2 and T-84 cells was much higher than that in the HeLa cells. Conclusion High level of MIP-3α mRNA could be found in mucosal epithelial cells and difference in transcription level of MIP-3α may exist in epithelial cells from different mucosa.
