SNHG6通过上调ZEB1表达促进食管鳞状细胞癌TE1细胞的侵袭和转移

SNHG6 promotes invasion and metastasis of esophageal squamous cell carcinoma TE1 cells via upregulating ZEB1

目的:探究长链非编码RNA(long non-coding RNA,lncRNA)核仁小分子RNA宿主基因6(small nuclear RNA host gene 6, SNHG6)促进食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)细胞侵袭和转移的作用及其分子生物学机制。方法:使用实时定量聚合酶链式反应(real time quantitative polymerase chain reaction,qPCR)检测36例ESCC组织及其癌旁组织(标本收集自河北医科大学第四医院2019年2月至8月外科手术的ESCC患者)中SNHG6表达水平;使用反转录聚合酶链式反应(reverse transcription polymerase chain reaction,RT-PCR)检测ESCC细胞系(TE1、Yes-2、Eca9706和Kyse150)中SNHG6表达水平,选用SNHG6高表达的TE1细胞,通过转染SNHG6-siRNA敲低该细胞中SNHG6表达;通过集落形成实验、划痕愈合实验和Transwell侵袭实验分别检测SNHG6敲低前后TE1细胞克隆形成、迁移和侵袭能力的变化;采用蛋白质免疫印迹法(Western blotting)检测SNHG6敲低前后TE1细胞中锌指蛋白E-盒结合同源异形盒-1(zinc finger E-box binding homeobox factor 1, ZEB1)、MMP-2和MMP-9蛋白表达的变化。结果:SNHG6在ESCC组织和细胞系中均呈高表达状态(均P<0.01),且在TE1细胞中高表达最为显著。转染SNHG6-siRNA后TE1细胞中SNHG6表达水平显著降低(P<0.01),si-SNHG6-1和si-SNHG6-2组TE1细胞的克隆形成、迁移和侵袭能力均显著低于对照组(均P<0.01),两组细胞中MMP-2、MMP-9和ZEB1表达水平均显著低于对照组(均P<0.05)。结论:ESCC组织中呈高表达的SNHG6可促进TE1细胞的恶性生物学行为,其机制可能涉及ZEB1表达的上调。

Objective: To explore the roles and mechanisms of long non-coding RNA(lncRNA) small nucleolar RNA host gene 6(SNHG6) in promoting invasion and metastasis of esophageal squamous carcinoma(ESCC). Methods: Real time quantitative polymerase chain reaction(qPCR) was used to detect the expression of SNHG6 in ESCC and matched para-carcinoma tissues. Reverse transcription-polymerase chain reaction(RT-PCR) was performed to detect the expression of SNHG6 in ESCC cell lines(TE1, Yes-2,Eca9706 and Kyse150). Then, TE1 cell line which harbored highest expression of SNHG6 was used in following experiments. siRNAs were used to knock down the expression of SNHG6. Clone formation, wound-healing and transwell assay were used to detect the abilities of proliferation, migration and invasion of TE1 cells, respectively. Western blotting was used to detect the expressions of MMP-2,MMP-9 and ZEB1 protein before and after knockdown of SNHG6 in TE1 cells. Results: SNHG6 was highly expressed in ESCC tissues,compared to para-carcinoma tissues(P<0.01). The expression of SNHG6 was significantly decreased after transfection of SNHG6-siRNA(all P<0.01). The abilities of proliferation, migration and invasion of TE1 cells in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group(all P<0.01). The expressions of ZEB1, MMP-2 and MMP-9 in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group(all P<0.05). Conclusion: SNHG6 is highly expressed in ESCC tissues and promotes the malignant biological behavior of ESCC cells. Its mechanism of promoting the occurrence and development of ESCC may be related to the upregulation of ZEB1 expression.