长链非编码RNA XLOC_005009对食管鳞状细胞癌生物学特性的影响

Effect of lncRNA XLOC_005009 on biological properties of esophageal cancer cell

目的:探讨长链非编码RNA XLOC_005009基因对食管鳞状细胞癌(ESCC)的体外增殖、迁移、侵袭和细胞周期与细胞凋亡等生物学特性的影响。方法:收集2015-2016年河北医科大学第四医院肿瘤研究所57例ESCC患者手术切除的癌组织及癌旁组织标本。应用实时荧光定量PCR检测XLOC_005009基因在人ESCC细胞系Eca109、Kyse170与ESCC组织及其癌旁组织的表达,构建pc DNA3.1-XLOC_005009过表达质粒并转染Eca109、Kyse170细胞,应用MTS法、克隆形成实验、划痕实验、Transwell小室、流式细胞术分别检测转染前后细胞增殖、克隆形成、迁移、侵袭、细胞周期与凋亡的变化。结果:XLOC_005009 m RNA在食管癌组织中的表达明显低于癌旁组织(0.06±0.06 vs 0.21±0.19,P<0.05),且食管癌细胞XLOC_005009 m RNA表达亦均低于对照组(P<0.05)。转染过表达质粒的Eca109和Kyse170细胞XLOC_005009基因表达显著高于对照组(Eca109细胞:039±0.17 vs0.02±0.00;Kyse170细胞:0.35±0.08 vs 0.01±0.01,均P<0.05)。与对照组相比,XLOC_005009基因过表达Eca109和Kyse170细胞增殖能力细胞显著减弱(P<0.05);克隆形成率显著减少(P<0.05);穿膜细胞数显著减少[Eca109细胞:(146.40±34.47)vs(193.00±26.33);Kyse170细胞:(157.80±32.51)vs(269.00±29.89),均P<0.05];Eca109细胞迁移率无显著变化,Kyse170细胞迁移率明显减小(P<0.05);S期细胞比例增加,细胞凋亡影响不明显。结论:XLOC_005009低表达与食管癌的发生发展密切相关,XLOC_005009过表达可抑制食管癌细胞的体外增殖、侵袭与迁移能力。

Objective： To explore effect of long non coding RNA （lonRNA） of XLOC_005009 gene on biological properties, proliferation, migration, invasion, cell cycle and apoptosis in vitro, of esophageal cancer cell. Methods：The surgically resected cancer and para-cancerous tissues of 25 patients with esophageal cancer who hospitalized in the Tumor Institute of the 4 th Hospital of Hebei Medical University during 2015 to 2016 were collected. Expressions of XLOC_005009 gene in human esophageal cancer Eca109 and Kyse170 line cells, the esophageal cancer and para- cancerous tissues were detected by RT-PCR. pcDNA3.1-XLOC_005009 over-expression plasmid was structured and transfected into the Eca109 and Kyse170 cells. MTS , colony forming, scratching, Transwell chamber and flow cytometry assays were used respectively to check proliferation, cloning efficiency, migration, invasion, cell cycle and apoptosis of the cells, before and after transfection of the over-expression plasmid. Results： Expression of XLOC_005009 mRNA in the esophageal cancer tissue was obviously lower than that in the para-cancerous normal tissue （0.06±0.06 vs 0.21±0.19, P〈0.05）, and expressions of XLOC_005009 mRNA in the esophageal cancer cells were all lower than that in the control group. In the Eca109 and Kyse170 cells transfected with the over-expression plasmid, expressions of XLOC_005009 gene were higher than those in the control groups （Eca109 cell： 039±0.17 vs 0.02±0.00; Kyse170 cell： 0.35±0.08 vs 0.01±0.01, all P〈0.05）. And comparing with the control group, proliferation abilities of the Eca109 and Kyse170 cells with over-expression of XLOC_005009 gene were significantly weakened （P〈0.05）, their cloning efficiency evidently reduced, transmembrane cell numbers of the cells remarkably decreased （Eca109 cell： 146.40±34.47 vs 193.00±26.33; Kyse170 cell：157.80±32.51 vs 269.00±29.89, all P〈0.05）, migration efficiency of the Eca109 cell didn’t markly change, but that of the Kyse170 cell obviously reduced; S phase cell ra- tio of the cells increased; effecting on apoptosis of the cells was not obvious. Conclusion： Low-expression of XLOC_005009 might be closely related to occurrence and development of the esophageal cancer. Over-expression of XLOC_005009 could inhibit proliferation, invasion and migration in vitro of the esophageal cancer cells.