ABSTRACT
To investigate various microRNAs (miRNAs) expression levels in cancer associated fibroblasts (CAFs) of gastric cancer,and to explore the effects of miRNA214 in CAFs on the invasion and migration of gastric cancer cells.Methods The primary CAFs were isolated from human gastric tumor tissues,while the normal fibroblasts (NFs) were from adjacent normal gastric mucosa tissues.Real-time quantitative PCR (RT-PCR) was performed to detect the expression level of miRNAs in CAFs and NFs.The interaction model in vitro between CAFs and MGC-803 (a gastric cancer cell line) was established by Transwell chamber,and then the effects of miRNA-214 in CAFs on MGC-803 invasion and migration were analyzed.Results It was found that the expression level of 18kinds of miRNAs in CAFs was different.Moreover,the expression level of miRNA-214 in CAFs was most observably decreased compared with NFs.Further studies indicated that over-expression of miRNA-214 in CAFs could markedly inhibit the migration and invasion ability of gastric cancer cells in vitro.Conclusion The expression of miRNAs was different between CAFs and NFs,and the miRNA-214 expression in CAFs is markedly reduced compared with NFs.The over-expression of miRNA-214in CAFs could significantly inhibit the migration and invasion ability of gastric cancer cells.Therefore,miRNA-214 in CAFs might act as a tumor suppressor in gastric cancer progression.
ABSTRACT
Objective To observe the changes of microRNA‐214 (miR‐214) expression in rat pulmonary artery smooth mus‐cle cells (PASMCs) induced by different hypoxia time ,and lay the foundation to explore the effect and mechanism of regulation of miR‐214 on PASMCs proliferation .Methods The primary cultured PASMCs were cultured under hypoxic 0 h ,6 h ,12 h ,24 h ,48 h ,respectively .The real time quantitative PCR was used to detect miR‐214 expression in each group PASMCs .Results The ex‐pression of miR‐214 in hypoxia group PASMCs was sustained as time increased ,apart from hypoxic hypoxia 6h group and 0h group ,the expression of miR‐214 was no significant difference (P>0 .05);the expression of miR‐214 among other groups PASMCs was significantly different (P<0 .05) .Conclusion The expression of miR‐214 in PASMCs increased after induction of hypoxia .We speculated that miR‐214 may be involved in the regulation of hypoxia induced PASMCs proliferation .
ABSTRACT
Objective To explore the inhibitory effect of anti-miRNA-214 oligonucleotide on leukemia U937 cells.Methods U937 cells were transfected with anit-miRNA-214 oligonucleotide,cell viability was analyzed by MTT assay.Apoptosis was detected by flow cytometry.The expression of miRNA214 in the U937 cells were measured by real-time PCR.Results The MTT result showed that the growth of U937 cells treated with AMO-miRNA-214 in 24 h,48 h,72 h was obviously inhibited (0.812±0.001,0.770±0.002,0.541±0.001),compared with those in control groups (randomized control 1.011±0.002,1.112±0.003,1.111±0.003,blank control 1.112±0.001,1.023±0.001,1.101±0.001),the differences had statistical significance (F =2.782,3.659,2.735,P =0.021,0.018,0.036).The flow cytometry results showed that the apoptosis detected in AMO-miRNA-214 group at 48 h,72 h (15.12±0.02,19.14±0.01) had significant differences compared with those in control groups (randomized control 2.04±0.02,2.45±0.03,blank control 1.19±0.02,2.02±0.01) (F =3.683,3.762,P =0.013,0.015).The expression of miRNA-214 was downregulated significantly in U937 cells after treated with oligonucleotide (31.1±0.2) compared with those in control groups (randomized control 25.8±0.1,blank control 25.6±0.2) (P ＜ 0.05).Conclusion Targeted inhibition of miRNA-214 with oligonucleotide can suppress U937 cells growth and induce apoptosis.