南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (11): 1615-1621.doi: 10.12122/j.issn.1673-4254.2020.11.12

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沉默长链非编码RNA HIF1A-AS2可抑制宫颈癌细胞的增殖、侵袭及迁移

邓 蓉，张福云，雷福珍，刘文涓，刘丝荪，王文华   

  • 出版日期:2020-11-20 发布日期:2020-11-23

Silencing long non-coding RNA HIF1A-AS2 inhibits proliferation, invasion and migration of cervical cancer cells in vitro

  • Online:2020-11-20 Published:2020-11-23

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摘要： 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。

关键词: 宫颈癌, 长链非编码RNA HIF1A-AS2, 上皮间质转换, 肿瘤干细胞

Abstract: 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). Conclusion Silencing HIF1A-AS2 can inhibit proliferation, invasion and migration of cervical cancer cells in vitro.

Key words: cervical cancer, long non-coding RNA HIF1A-AS2, epithelial-mesenchymal transition, tumor stem cell