摘要
目的探讨长链非编码RNA RP5-919F19对胃癌细胞增殖和侵袭迁移的影响。方法采用实时定量PCR(qPCR)检测RP5-919F19在胃癌细胞(MGC-803、SGC-7901和BGC-823)以及人正常胃黏膜上皮细胞(GES-1)中的表达情况;构建干扰RP5-919F19表达的小干扰RNA序列(siRNA-1~3)并采用脂质体法转染胃癌细胞,根据转染情况分为转染干扰效率最高siRNA序列的干扰组、转染阴性对照(si-NC)序列的NC组和不行任何转染的空白对照(Blank)组,采用qPCR、MTT比色法、划痕实验和Transwell小室实验检测RP5-919F19水平、增殖活力、划痕愈合率和穿膜细胞数,qPCR和Western blotting检测基质金属蛋白酶(MMP)-2/9水平。结果胃癌细胞的RP5-919F19水平均高于GES-1细胞,其中SGC-7901细胞的水平最高(P<0.05);与转染si-NC序列的SGC-7901细胞相比,转染siRNA-1~3序列的RP5-919F19水平均降低,其中序列siRNA-2对应的RP5-919F19水平最低(P<0.05);选取SGC-7901细胞和siRNA-2序列进行后续分组实验。与Blank组和NC组相比,干扰组的RP5-919F19水平降低且处理48~72 h后细胞增殖活力减弱(P<0.05)。干扰组的划痕愈合率和穿膜细胞数分别为(18.971±3.869)%和(69.573±9.601)个,均低于Blank组的(61.887±4.421)%和(152.707±23.176)个及NC组的(59.216±4.557)%和(161.820±15.442)%(P<0.05)。与Blank组和NC组相比,干扰组MMP-2和MMP-9的mRNA和蛋白水平均降低(P<0.05)。Blank组和NC组上述指标的差异无统计学意义(P>0.05)。结论RP5-919F19在胃癌细胞中高表达且发挥促癌作用,通过外源siRNA手段下调其表达可抑制SGC-7901细胞的增殖和迁移侵袭能力,可能与抑制MMP-2和MMP-9表达有关,是胃癌潜在的治疗靶点。
Objective To investigate the effect of long non-coding RNA RP5-919F19 on the proliferation,invasion and migration of gastric cancer cells.Methods The expression of RP5-919F19 in gastric cancer cells(MGC-803,SGC-7901 and BGC-823)and human normal gastric mucosa epithelial cells(GES-1)was detected by real-time quantitative PCR(qPCR).The small interfering RNA(siRNA-1-3)targeting the RP5-919F19 expression was constructed and transfected into gastric cancer cells by liposome method.According to the transfection conditions,cells were divided into Interference group with siRNA sequence of the highest interference efficiency,NC group with negative control(si-NC)sequence and Blank group without any transfection.RP5-919F19 level,proliferation activity,scratch healing rate and number of membrane-penetrating cells were detected by qPCR,MTT colorimetry,scratch test and Transwell chamber test.Levels of matrix metalloproteinase(MMP)-2/9 were detected by qPCR and Western blotting.Results The RP5-919F19 levels of gastric cancer cells were higher than that of GES-1 cells,of which SGC-7901 cells had the highest level(P<0.05).Compared with SGC-7901 cells transfected with si-NC sequence,RP5-919F19 levels of siRNA-1-3 sequence were lower,and the RP5-919F19 levels of siRNA-2 sequence was the lowest(P<0.05).SGC-7901 cells and siRNA-2 sequence were selected for subsequent experiments.Compared with Blank group and NC group,the RP5-919F19 level of the Interference group decreased and the cell proliferation activity decreased after 48-72 h of treatment(P<0.05).The scratch healing rate and number of membrane-penetrating cells in Interference group were(18.971±3.869)%and 69.573±9.601,lower than(61.887±4.421)%and 152.707±23.176 in Blank group and(59.216±4.557)%and 161.820±15.442 in NC group(P<0.05).Compared with Blank group and NC group,the mRNA and protein levels of MMP-2 and MMP-9 in Interference group decreased(P<0.05).There was no significant difference between Blank group and NC group on the above indices(P>0.05).Conclusion RP5-919F1
作者
韩帅
马俊霞
黄加国
HAN Shuai;MA Junxia;HUANG Jiaguo(Department of Gastroenterology, Jinan People's Hospital, Jinan 271199, China)
出处
《临床肿瘤学杂志》
CAS
北大核心
2020年第5期399-404,共6页
Chinese Clinical Oncology
