长链非编码RNA H19对胃癌细胞侵袭、迁移的影响及其机制

Influence of lncRNA H19 on invasion and migration of gastric cancer cells

目的探讨长链非编码RNA H19对胃癌细胞侵袭和迁移的影响,并初步探讨其作用机制。方法将处于对数生长期的MKN28细胞分为MKN28/H19组、MKN28/H19mut组和空白对照1组,MKN28/H19组、MKN28/H19mut组采用脂质体法转染全长H19 cDNA和突变H19 cDNA(不含第一个内含子),空白对照1组不转染。将对数生长期的BGC-823细胞分为BGC-823/siH19组、BGC-823/NC组和空白对照2组,BGC-823/siH19组、BGC-823/NC组同法分别转染H19 siRNA、H19 siRNA无关序列,空白对照2组不转染。各组均转染12 h。采用实时荧光定量PCR法检测各组H19相对表达量;采用Tranwell细胞侵袭试验观察各组细胞侵袭能力(以下室OD570值表示);采用Tranwell细胞迁移试验观察各组细胞迁移能力(以下室OD570值表示);采用Western blotting法检测各组H19共结合蛋白CPT1C相对表达量。结果转染12 h后MKN28/H19组H19相对表达量、侵袭能力、迁移能力、CPT1C相对表达量均高于MKN28/H19mut组、空白对照1组(P均<0.05),MKN28/H19mut组上述指标与空白对照1组相比,P均>0.05。转染12 h后BGC-823/siH19组H19相对表达量、侵袭能力、迁移能力、CPT1C相对表达量均低于BGC-823/NC组、空白对照2组(P均<0.05),BGC-823/NC组上述指标与空白对照2组相比,P均>0.05。结论上调胃癌细胞H19表达可以提高胃癌细胞侵袭和迁移能力,下调胃癌细胞H19表达可以抑制胃癌细胞侵袭和迁移能力。H19可能是通过调节CPT1C蛋白表达调节胃癌细胞侵袭和迁移能力。

Objective To investigate the effects of long non-coding RNA H19 on the migration and invasion of gastric cancer cells, and to explore its possible mechanism. Methods The MKN28 cells in the logarithmic growth phase were di- vided into the MKN28/H19 group, the MKN28/H19mut group, and the blank control group 1. The full-length H19 cDNA and the mutant H19 cDNA （without the first exon） were transfeeted into the MKN28/I-I19 group and MKN28/H19mut group by the liposome transfection, respectively. The BGC-825 cells in the logarithmic growth phase were divided into the BGC-823/siH19 group, BGC-823/NC group, and the blank control group 2. The H19 siRNA and H19 siRNA-negative control were transfected into the BGC-823/siH19 group and BGC-823/NC group, respectively. At 12 h after transfection, the real-time fluorescent quantitative PCR was applied to detect the expression of lncRNA H19 in each group. The invasion capability was analyzed by the Transwell invasion assay （ ODs70 ） and the migration capability was analyzed by the Transwell migration assay （OD570 ）. The relative expression level of H19 co-expressed protein CPT1C was detected by Western blot- ting. Results At 12 h after the transfection, the relative expression level of H19, the migration capability, the invasion capability, and the the relative expression level of CPT1C in the MKN28/H19 group were much higher than those in the MKN28/H19mut and the blank control group 1 （ all P 〈 0.05）, but there were no differences between the MKN28/H19mut and the blank control group 1 （P 〉0.05）. Moreover, at 12 h after the transfection, the relative expressibn level of H19, the migration capability, the invasion capability, and the the relative expression level of CPT1C in the BGC-823/siH19 group were significantly lower than those in the BGC-823/NC and the blank control group 2 （ all P 〈 0.05）, but there were no differences between the BGC-823/NC and the blank control group 2 （ P 〉 0, 05 ）. Conclusions Up-regulating the HI9 could enhance the migration and invasion of gastric cancer ceils, while down-regulating the H19 could inhibit the migration and invasion of gastric cancer ceils. H19 may participate in the migration and invasion of gastric cancer cells by regulating the protein expression of CPT1C.