Chen Mingjiu, Xia Zhenkun, Chen Chen, Hu Wen, Yuan Yunchang
Department of Thoracic Surgery, The Second Xiangya Hospital of Central South University, Changsha, People's Republic of China.
Anticancer Drugs. 2018 Sep;29(8):767-773. doi: 10.1097/CAD.0000000000000645.
To investigate effect of long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) on epithelial-to-mesenchymal transition (EMT) of esophageal cancer (EC) and role of enhancer of zeste homolog 2 (Ezh2)-Notch1 signaling pathway in the process. The expression of MALAT1 was determined in four EC cell lines by real-time PCR. TE-1 and EC109 cells were transfected with sh-MALAT1 to inhibit expression of MALAT1 or transfected with pcDNA3.1-Ezh2 to overexpress Ezh2. Invasion and migration assays were conducted to analyze cell metastasis, and expressions of Ezh2-Notch1 signaling-related proteins as well as EMT related proteins were determined using both real-time PCR and western blot. MALAT1 was significantly up-regulated in all EC cell lines compared with the normal cells. Silencing MALAT1 using shRNA could significantly inhibit cell viability (reduced almost 30% of cell viability compared with the control), invasion (reduced almost 30% of cell migration compared with the control), and migration (reduced almost 50% of cell migration compared with the control) of both TE-1 and EC109 cells (P<0.05). Meanwhile, expression of Ezh2, Notch1, Hes1, MMP-9, and Vimentin was significantly decreased and expression of E-cadherin was significantly increased when cells were transfected with sh-MALAT1 compared with the nontransfected cells (P<0.05). However, when cells were cotransfected with both sh-MALAT1 and pcDNA3.1-Ezh2, the protein expression changes induced by sh-MALAT1 were recovered. MALAT1 could affect EMT and metastasis of EC cells through Ezh2-Notch1 signaling pathway. This study can give deeper understandings of the role of MALAT1 in EC and may provide some new directions for treatment of patients with EC.
探讨长链非编码RNA转移相关肺腺癌转录本1（MALAT1）对食管癌（EC）上皮-间质转化（EMT）的影响以及zeste同源物2（Ezh2）-Notch1信号通路在此过程中的作用。采用实时定量聚合酶链反应（PCR）检测4种EC细胞系中MALAT1的表达。分别转染靶向MALAT1的短发夹RNA（sh-MALAT1）抑制TE-1和EC109细胞中MALAT1的表达，或转染pcDNA3.1-Ezh2过表达Ezh2。进行侵袭和迁移实验分析细胞转移能力，采用实时定量PCR和蛋白质免疫印迹法检测Ezh2-Notch1信号通路相关蛋白以及EMT相关蛋白的表达。与正常细胞相比，所有EC细胞系中MALAT1均显著上调。shRNA沉默MALAT1可显著抑制TE-1和EC109细胞的活力（与对照组相比，细胞活力降低近30%）、侵袭能力（与对照组相比，细胞迁移减少近30%）和迁移能力（与对照组相比，细胞迁移减少近50%）（P<0.05）。同时，与未转染细胞相比，转染sh-MALAT1的细胞中Ezh2、Notch1、Hes1、基质金属蛋白酶-9（MMP-9）和波形蛋白的表达显著降低，E-钙黏蛋白的表达显著增加（P<0.05）。然而，当细胞同时转染sh-MALAT1和pcDNA3.1-Ezh2时，sh-MALAT1诱导的蛋白表达变化得以恢复。MALAT1可能通过Ezh2-Notch1信号通路影响EC细胞的EMT和转移。本研究有助于深入了解MALAT1在EC中的作用，可能为EC患者的治疗提供新的方向。
