[关键词]
[摘要]
目的:探讨长链非编码RNA(long non-coding RNA, lncRNA)NUP50-AS1 在食管鳞状细胞癌(esophageal squamous cell cancer, ESCC)组织及细胞株中的表达及其对人食管癌Eca109 细胞增殖、迁移和侵袭的影响。方法:选取自2015 年1 月至2016 年12 月河北医科大学第四医院生物标本库的49 例ESCC手术患者的癌组织和相应癌旁组织,qRT-PCR检测ESCC癌组织、癌旁组织及5 种食管癌细胞株(TE1、TE13、Eca109、Kyse150 和Kyse170)中NUP50-AS1 表达水平。shRNA 转染NUP50-AS1 后,选用sh2 -NUP50-AS1 进行后续功能实验。采用MTS法、克隆形成实验检测敲减NUP50-AS1 表达对Eca109 细胞增殖的影响,划痕实验检测敲减NUP50-AS1 表达对细胞迁移的影响,Transwell 小室实验检测敲减NUP50-AS1 表达对细胞侵袭的影响。结果:NUP50-AS1 在ESCC组织中的相对表达量显著高于癌旁组织(2.003±0.870 vs 1.000±0.000,P<0.05);NUP50-AS1 在ESCC组织中的表达水平与淋巴结转移及TNM分期相关(均P<0.01),NUP50-AS1 在5 株食管癌细胞系中相对表达量均明显上调(P<0.05),其中Eca109 细胞的NUP50-AS1 表达水平最高。转染后,sh2-NUP50-AS1 转染组干扰效率最高,敲低NUP50-AS1 可明显抑制Eca109 细胞增殖、迁移和侵袭能力。结论:ESCC组织中lncRNA NUP50-AS1 表达明显高于癌旁组织,且与癌症分期和淋巴结转移有关,敲减其表达明显抑制食管癌细胞增殖、迁移和侵袭能力,NUP50-AS1 的高表达可能与ESCC的发生发展密切相关。
[Key word]
[Abstract]
Objective: To investigate the expression of long non-coding RNA NUP50-AS1 (lncRNA NUP50-AS1) in esophageal squamous cell carcinomas (ESCC) tissues and cell lines, and to explore its effect on proliferation, migration and invasion of human esophageal cancer Eca109 cells. Methods: 49 pairs of ESCC tissues and corresponding para-cancerous tissues obtained from the Biological Specimen Base of the Fourth Hospital of Hebei Medical University during Jan. 2015 to Jan. 2016 were used in this study. qRT-PCR method was applied to detect the expression of NUP50-AS1 in collected tissues samples and five esophageal cancer cell lines (TE1,TE13, Eca109, Kyse150 and Kyse170). ShRNAs were transiently transfected into Eca109 cells to interfere the expression of NUP50-AS1 gene, and finally, sh2-NUP50-AS1 was used for the following experiments. The effect of NUP50-AS1 gene knockdown on the proliferation of Eca109 cells was detected by MTS and colony formation assay; the effect of NUP50-AS1 gene knockdown on the migration of Eca109 cells was detected by scratch test, and the effect on cell invasion was detected by Transwell assay. Results: The expression of NUP50-AS1 in ESCC was correlated with the lymphnode metastasis and TNM stage (all P<0.01). The expression of NUP50-AS1 in ESCC tissues was significantly higher than that in corresponding normal tissues (2.003±0.870 vs 1.000±0.000, P<0.05). The expression of NUP50-AS1 in five esophageal cancer cell lines was significantly up-regulated (P<0.05), and it had the highest expression in Eca109 cell line. After transfection, sh2-NUP50-AS1 had the highest transfection efficiency, and knocking down NUP50-AS1 gene significantly inhibited the proliferation, invasion and migration of the Eca109 cells. Conclusion: The expression of lncRNA NUP50-AS1 in ESCC tissues was significantly higher than that in the para-cancerous tissues, and correlated with the TNM stage and lymphnode metastasis. The down-regulation of NUP50-AS1 inhibited the proliferation, invasion and migration of esophageal cancer cells.The high expression of NUP50-AS1 gene may be closely related to the occurrence and development of ESCC.
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[基金项目]
国家自然科学基金资助项目(No. 81572441);河北省医学科学研究重点课题资助项目(No. 20170700)