miR-449c在胃癌中的表达

Expression of miR-449c in Gastric Cancer

目的:检测胃癌组织中miR-449c的表达情况,探讨其与胃癌发生及预后的关系。方法:选取32例胃癌患者,实时荧光定量PCR(q RT-PCR)法检测癌旁组织(NGT)、胃癌组织(GC)及不同细胞株中miR-449c表达水平,Lipofectamine 2000转染法对相关细胞株miR-449 c表达进行上调和下调,MTT方法检测细胞的生长活力变化,流式细胞术检测细胞凋亡情况;随访30个月,比较miR-449c低表达和高表达患者的生存率。结果:GC组织miR-449c表达(0.36±0.28)明显低于NGT组织miR-449c表达(1.01±0.14)(P<0.05),且AGS、SNU-1、SNU-5、SNU-16细胞miR-449c表达(0.39±0.08)、(0.33±0.07)、(0.25±0.06)、(0.22±0.04)均明显低于NGT组织(1.01±0.14)(P<0.05);miR-449c模拟物对SNU-16细胞进行转染,其miR-449 c表达水平(8.49±2.50)明显高于空质粒转染(P<0.05),第2、3 d细胞生长活力(0.80±0.10)、(0.90±0.12)明显低于空质粒转染(P<0.05),凋亡率(80.12±9.01)明显高于空质粒转染(P<0.05);miR-449c抑制物对AGS细胞进行转染,其miR-449c表达水平(3.00±0.50)明显低于空质粒转染(P<0.05),第2、3 d细胞生长活力(1.20±0.14)、(1.70±0.18)明显高于空质粒转染(P<0.05),凋亡率(8.00±1.00)明显低于空质粒转染(P<0.05);随访30个月,miR-449c高表达患者生存率50.00%明显高于miR-449c低表达者(P<0.05)。结论:miR-449c在胃癌组织中低表达,抑制细胞生长,促进细胞凋亡,其低表达可能与胃癌患者低生存率有关。

Objective To detect the miR-449c expression in gastric cancer(GC)and discuss the relationship between miR-449c expression and carcinogenesis and prognosis of GC.Methods A total of 32 patients with GC were selected as researching subjects.Real-time fluorescence quantitative PCR(Real-time PCR)was used to detect the expression of miR-449c in normal gastric tissue(NGT),gastric cancer tissues(GC)and GC cell lines.miR-449c mimics and inhibitors were trasfected into GC cells according to the instructions of Lipofectamine 2000.Cell viability was detected by MTT and cell apoptosis was detected by FACS.The survival rate was analyzed between the patients with high expression of miR-449c and the patients with low expression of miR-449c.Results The results showed that the expression level of miR-449c of GC(0.36±0.28)was significantly lower than that of NGT group(1.01±0.14,P<0.05).The expression level of miR-449c in AGS,SNU-1,SNU-5 and SNU-16 cell lines(0.39±0.08,0.33±0.07,0.25±0.06,0.22±0.04,respectively)was significantly lower than that in NGT(P<0.05).The expression level of miR-449c(8.49±2.50)in SNU-16 cell line transfected with miR-449c mimics was significantly higher than that of SNU-16 cells transfected with scrambled sequences(scr)(P<0.05).Cell viability at second day(0.80±0.10)and third day(0.90±0.12)was significantly lower than that of SNU-16 cells transfected with scr(P<0.05).The apoptosis rate(80.12±9.01)was significantly higher than that of SNU-16 cells transfected with scr(P<0.05).The expression level of miR-449c in AGC cells transfected with miR-449c inhibitor(3.00±0.50)was significantly lower than that of AGC cells transfected with scr(P<0.05).Cell viability at second day(1.20±0.14)and third day(1.70±0.18)was significantly higher than that of AGC cells transfected with scr(P<0.05).The apoptosis rate(8.00±1.00)was significantly lower than that of AGC cells transfected with scr(P<0.05).Followed-up for 30 months,survival rate(50%)in the patients with high miR-449c expression level was significantly higher than that of low miR-449c expression level(P<0.05).Conclusion miR-449c expressed lower in GC tissue.It could inhibit cell growth and promote apoptosis.miR-449c expression was closely correlation with the survival rate of the GC patients.