Objective: To explore the inhibitory effects of silencing long non-coding RNA (LncRNA) HIF1A-AS2 on epithelialmesenchymal transition (EMT) and tumor stem cell-like phenotype in cervical cancer cells.
Methods: We designed 3 shRNA constructs for silencing HIF1A-AS2 in CaSki cells, and the shRNA with the strongest interference effect was selected for subsequent experiment. CaSki cells were transfected with shRNA-NC or Sh-HIF1A-AS2, and the changes in cell viability, invasion ability, EMT, expressions of EMT-related proteins, formation of cell spheres and expressions of stem cell markers were detected.
Results: Transfection with shRNA-NC and Sh-HIF1A-AS2 did not significantly affected the viability of CaSki cells (P > 0.05). Compared with the cells transfected with shRNA-NC, the cells transfected with Sh- HIF1A-AS2 showed significantly reduced invasion ability, expressions of vimentin N-cadherin, and cell sphere formation ability. HIF1A-AS2 silencing obviously lowered the rate of ABCG2-positive cells, significantly reduced the mRNA and protein expressions of Nanog, OCT4, and SOX2, and strongly enhanced the expression of E-cadherin in CaSki cells (P < 0.05).
Conclusions: Silencing HIF1A-AS2 can inhibit proliferation, invasion and migration of cervical cancer cells in vitro.
目的: 探究长链非编码RNA HIF1A-AS2诱导宫颈癌细胞的上皮间质转换和肿瘤干细胞样特性的机制。
方法: 设计3种可以沉默HIF1A-AS2的shRNA转染caski细胞株，选出干扰效果最好的shRNA进行后续研究；将caski细胞分为：Control组、shRNA-NC组和Sh-HIF1A-AS2组；检测各组细胞活力、侵袭能力、上皮间质转换及相关蛋白表达情况、肿瘤细胞成球情况和干细胞标记物情况；
结果: 各组细胞活性差异无统计学意义（P > 0.05）；干扰结果确认使用HIF1A-AS2-shRNA1为shRNA；相比Control组，shRNA-NC组所有指标均无显著改变（P > 0.05）；相比shRNA-NC组，Sh-HIF1A-AS2组单位面积侵袭细胞数目、Vimrntin和N-cadherin、细胞成球数目、成球细胞直径、ABCG2阳性细胞率、Nanog、OCT4、SOX2蛋白和mRNA相对表达均显著降低，E-cadherin相对表达显著升高（P < 0.05）。
结论: 沉默长链非编码RNA HIF1A-AS2可以抑制宫颈癌细胞的增殖、侵袭及迁移能力。
Keywords: cervical cancer; epithelial-mesenchymal transition; long non-coding RNA HIF1A-AS2; tumor stem cell.