长链非编码RNA lucat1对胃癌细胞侵袭和迁移能力的影响及其机制

Effects of long non-coding RNA lucat1 on invasion and migration of gastric cancer cells

目的探讨长链非编码RNA(lncRNA)lucat1对胃癌细胞侵袭和迁移能力的影响及其机制。方法 (1)采用实时荧光定量PCR法检测正常胃黏膜细胞GSE-1和胃癌细胞SGC-7901、BGC-823中lncRNA lucat1和对甲酰肽受体2(FPR2)mRNA。(2)将体外培养的胃癌细胞SGC-7901、BGC-823分为观察组和对照组。观察组用Lipofectamine2000转染lucat1 shRNA,对照组不转染。转染48 h,采用实时荧光定量PCR法检测两组FPR2 mRNA相对表达量。(3)将体外培养的胃癌细胞BGC-823分为A、B、C组。A组转染lucat1 shRNA,B组转染lucat1 shRNA和FPR2过表达质粒,C组不转染。转染48 h,用Transwell小室实验和划痕实验观察各组细胞侵袭和迁移能力,结果分别用穿膜细胞数和细胞迁移距离表示。结果 (1)SGC-7901、BGC-823、GES-1细胞lncRNA lucat1相对表达量分别为3.651±0.332、3.023±0.331、1.000±0.098,FPR2 mRNA相对表达量分别为3.354±0.433、3.425±0.354、1.000±0.277。SGC-7901、BGC-823细胞lucat1、FPR2 mRNA相对表达量均明显高于GES-1细胞(P均<0.05)。(2)观察组、对照组SGC-7901细胞FPR2 mRNA相对表达量分别为1.00±0.05、0.48±0.04,BGC-823细胞FPR2mRNA相对表达量分别为1.00±0.04、0.48±0.04,两组比较P<0.05。(3)Transwell小室实验A、B、C组穿膜细胞数分别为(143±7)、(194±9)、(233±8)个,三组穿膜细胞数两两比较P均<0.05。划痕实验A、B、C组细胞迁移距离分别为(0.342±0.027)、(0.534±0.021)、(0.652±0.029)mm,三组细胞迁移距离两两比较P均<0.05。结论 lncRNA lucat1可促进胃癌细胞侵袭和迁移能力,其机制可能与上调FPR2表达有关。

Objective To investigate the effects of long non-coding RNA lucat1 on the invasion and migration abilities of gastric cancer（GC） cells SGC-7901 and BGC-823 as well as its mechanism. Methods（1） The mRNA expression of formyl peptide receptor 2（FPR2） and lncRNA lucat1 in normal gastric mucosa cells（GSE-1） and GC cells（SGC-7901 and BGC-823） was detected by real-time fluorescent quantitative PCR;（2） SGC-7901 and BGC-823 cells were divided into the observation group and control group,respectively. The cells in the observation group were transfected with lucat1 shRNA by lipofectamine 2000 for 48 h,and the cells in the control group were untreated. The mRNA expression of FPR2 in the SGC-7901 and BGC-823 cells was examined by RT-PCR.（3） SGC-7901 and BGC-823 cells were divided into groups A,B,and C. Cells in the group A were transfected with lucat1 shRNA,cells in the group A were transfected with lucat1 shRNA and over-expressed FPR2,and cells in the group C were not treated. The changes of invasive and metastasis abilities in SGC-7901 and BGC-823 cells at 48 h after transfection were examined by Scratch test and Transwell assay,respectively.Results（1）The relative expression levels of lucat1 in the SGC-7901,BGC-823,and GES-1 cells were 3. 651 ± 0. 332,3. 023 ± 0. 331,and 1. 000 ± 0. 098,respectively. The relative expression levels of FPR2 mRNA were 3. 354 ± 0. 433,3. 425 ± 0. 354,and 1. 000 ± 0. 277,respectively. The relative expression of lucat1 and FPR2 mRNA in SGC-7901 and BGC-823 cells was significantly lower than that in GES-1 cells（P〈0. 05）.（2） The relative expression levels of FPR2 mRNA of SGC-7901 cells in the observation group and control group were 1. 00 ± 0. 05 and 0. 48 ± 0. 04,respectively; the relative expression levels of FPR2 mRNA in BGC-823 cells were 1. 00 ± 0. 04 and 0. 48 ± 0. 04,respectively,P〈0. 05.（3） The transmembrane cells in the groups A,B,and C were 143 ± 7,194 ± 9,and 233 ± 8; significant difference was found between every two group（all P〈0. 05）,with the lowest in group A,and highest in group C. The migration distance of groups A,B,and C were（0. 342 ± 0. 027）,（0. 534 ± 0. 021）,and（0. 652 ± 0. 029） mm,respectively,and significant difference was found between every two group（all P〈0. 05）,with the lowest in group A,and highest in group C.Conclusion LncRNA lucat1 can promote the invasive and metastasis of GC cells,and it is associated with FDR2.