[Knockdown of lncRNA CACNA1C-AS2 promotes epithelial-mesenchymal transition and enhances the proliferation, invasion and migration of human esophageal cancer cells].

Yuping Qin, Tong Cao, Ying Xu, Xu Jiang, Jia Ma, Li Yu
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Abstract

Objective To investigate the effect of calcium voltage gated channel subunit α 1C antisense RNA2 (CACNA1C-AS2) on malignant biological characteristics of esophageal cancer cells by regulating epithelial mesenchymal transition (EMT). Methods CACNA1C-AS2 expression levels in paracancerous tissues and esophageal cancer tissues were analyzed by TCGA database. Real-time quantitative PCR was used to detect the expression of CACNA1C-AS2 mRNA in esophageal cancer cells. Following the knockdown and high expression of CACNA1C-AS2 in esophageal cancer cells, the viability of the cells was tested by MTT assay and cell colony formation assay. TranswellTM chamber method was used to measure the invasion and longitudinal migration of the cells. The horizontal migration ability of the cells was evaluated by wound healing test. The apoptosis rates of cells were detected by flow cytometry. Western blot analysis was used to detect the expressions of N-cadherin, vimentin and slug. Results CACNA1C-AS2 expression levels were low in esophageal cancer tissues and cell lines. After knocking down the expression of CACNA1C-AS2 in EC-9706 cells and Eca-109 cells, the ability of invasion and migration and viability of esophageal cancer cells were significantly enhanced, and the apoptosis rates were decreased, while the expressions of N-cadherin, vimentin and slug were up-regulated. However, the results are opposite via the over-expression of CACNA1C-AS2. Conclusion CACNA1C-AS2 enhances the proliferation, invasion and migration of esophageal cancer cells by promoting EMT.

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[lncRNA CACNA1C-AS2敲低可促进人食管癌细胞的上皮-间质转化,增强其增殖、侵袭和迁移能力]。
目的探讨钙电压门控通道亚基α 1C反义RNA2 (CACNA1C-AS2)通过调控上皮间质转化(EMT)对食管癌细胞恶性生物学特性的影响。方法采用TCGA数据库分析CACNA1C-AS2在癌旁组织和食管癌组织中的表达水平。采用实时荧光定量PCR检测食管癌细胞中CACNA1C-AS2 mRNA的表达。在食管癌细胞中敲低并高表达CACNA1C-AS2后,采用MTT法和细胞集落形成法检测细胞活力。采用TranswellTM室法测定细胞的侵袭和纵向迁移。通过创面愈合试验评价细胞的水平迁移能力。流式细胞术检测细胞凋亡率。Western blot检测N-cadherin、vimentin、slug的表达。结果食管癌组织和细胞系中CACNA1C-AS2表达水平较低。在EC-9706细胞和Eca-109细胞中敲低CACNA1C-AS2的表达后,食管癌细胞的侵袭迁移能力和生存能力显著增强,凋亡率降低,N-cadherin、vimentin和slug的表达上调。然而,过表达CACNA1C-AS2则相反。结论CACNA1C-AS2通过促进EMT促进食管癌细胞的增殖、侵袭和迁移。
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