Effect of fibroblast activation protein expression silencing of mouse fibroblast cells on the proliferation of muse pancreatic cancer cells
10.3760/cma.j.issn.1674-1935.2012.04.008
- VernacularTitle:沉默小鼠成纤维细胞活化蛋白表达对原代胰腺癌细胞增殖及凋亡的影响
- Author:
Yebo SHAO
;
Dayong JIN
;
Yefei RONG
;
Xuefeng XU
- Publication Type:Journal Article
- Keywords:
RNA,small interfering;
Gene silence;
Fibroblast activation protein;
Pancreatic neoplasms;
Cell proliferation;
Apoptosis
- From:
Chinese Journal of Pancreatology
2012;12(4):242-245
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveSmall interfering RNA (siRNA) was used to silence the fibroblast activation protein4 (FAP) expression of mouse pancreatic cancer related fibroblast cells (mPCa-FCs-1212),and to observe the effects of mPCa-FCs-1212 silencing FAP gene on mouse pancreatic cancer cells (mPCa-1212) proliferation and apoptosis.MethodsThe small interfering RNA targeting FAP gene was designed; the recombinant siRNA plasmid siFAP and control plasmid siMOCK was constructed,which were transfected into mPCa-FCs-1212,respectively.The FAP mRNA and protein expression in transfected cells were examined by real-time PCR and Western blotting.The mPCa-1212 and transfected mPCa-FCs-1212 were co-cultured with a 1:1 ratio in vitro.The growth inhibitory rates and apoptosis rates of mPCa-1212 were detected by MTT assay and Annexin V-FTTC/PI staining and FCM assay.ResultsThe mRNA and protein expressions of FAP in siFAP transfected mPCa-FCs-1212 were significantly down-regulated when compared with that in siMOCK transfected mPCa-FCs-1212[0.584 ±0.029vs.1.052±0.281,P=0.0213; (27.18±3.23)％ vs.(61.58±4.72)％,P=0.0317].The mPCa-1212 was co-cultured with the mPCa-FCs-1212 transfected with siFAP or siMOCK for 3 d,and the inhibitory rates of mPCa-1212 were (23.02 ±3.32)％ and (1.11 ±0.23)％,and the apoptosis rates were (42.31 ±5.34)％ and (7.38 ± 2.09)％,the difference between the two groups was statistically significant (P =0.000).ConclusionsmPCa-FCs-1212 silencing FAP gene can inhibit the proliferation of mPCa-1212 in vitro and induce cell apoptosis,and may be a potential new approach to gene therapy.
