Dai Jing, Mu Jing-Wen, Mu Hong
Department of Stomatology, Jingzhou Central Hospital, The Second Clinical Medical College, Yangtze University, Jingzhou, Hubei 434020, P.R. China.
Department of Stomatology, Dongfang Hospital Beijing University of Chinese Medicine, Beijing 100078, P.R. China.
Oncol Lett. 2019 Mar;17(3):3330-3340. doi: 10.3892/ol.2019.9978. Epub 2019 Jan 28.
The present study aimed to investigate whether the long non-coding RNA (lncRNA) colorectal neoplasia differentially expressed (CRNDE) can promote the migration and invasion of human oral squamous cell carcinoma (OSCC) cells via the regulation of epithelial-mesenchymal transition (EMT). CAL-27 and SCC-15 cells were classified into a control group, a small interfering negative control (si-NC) group (cells transfected with control siRNA) and an si-CRNDE group (cells transfected with CRNDE siRNA). The expression of CRNDE in OSCC tissues and cell lines was detected by hybridization (ISH) and reverse transcription-quantitative polymerase chain reaction. An MTT assay was used to detect cell proliferation, flow cytometry was performed to determine cell apoptosis, wound-healing and Transwell assays were conducted to evaluate cell metastasis, and immunofluorescence staining and western blotting were performed to measure the expression of proteins associated with EMT. Tumor-bearing mouse models were established, and the tumor volumes were recorded. An immunohistochemical assay was performed to determine the expression of EMT-related proteins. CRNDE expression was increased in OSCC tissues and cell lines compared with that in normal tissues and cell lines. Compared with the control group, the si-CRNDE group displayed a reduction in the expression of CRNDE, in the proliferation, migration and invasion of cells, in the protein expression of N-cadherin, vimentin and Snail, and in the expression of proteins in the Wnt/β-catenin pathway. However, an increase was displayed in the apoptosis of cells and the expression of E-cadherin. Compared with the control group of tumor-bearing nude mice, the sh-CRNDE group demonstrated slowed tumor growth, reduced tumor weight and elevated E-cadherin, as well as reduced expression of N-cadherin, vimentin and Snail. In conclusion, silencing CRNDE may inhibit EMT, thus decreasing the migration and invasion of human OSCC cells by repressing the activation of the Wnt/β-catenin signaling pathway, thereby restricting cell growth and promoting cell apoptosis.
本研究旨在探讨长链非编码RNA（lncRNA）结直肠癌差异表达基因（CRNDE）是否能通过调控上皮-间质转化（EMT）来促进人口腔鳞状细胞癌（OSCC）细胞的迁移和侵袭。将CAL-27和SCC-15细胞分为对照组、小干扰阴性对照组（si-NC组，转染对照siRNA的细胞）和si-CRNDE组（转染CRNDE siRNA的细胞）。采用原位杂交（ISH）和逆转录-定量聚合酶链反应检测OSCC组织和细胞系中CRNDE的表达。采用MTT法检测细胞增殖，通过流式细胞术检测细胞凋亡，进行伤口愈合实验和Transwell实验评估细胞转移，采用免疫荧光染色和蛋白质印迹法检测与EMT相关的蛋白质表达。建立荷瘤小鼠模型并记录肿瘤体积。采用免疫组织化学法检测EMT相关蛋白的表达。与正常组织和细胞系相比，OSCC组织和细胞系中CRNDE表达升高。与对照组相比，si-CRNDE组CRNDE表达降低，细胞增殖、迁移和侵袭能力降低，N-钙黏蛋白、波形蛋白和Snail的蛋白表达降低，Wnt/β-连环蛋白通路中蛋白表达降低。然而，细胞凋亡和E-钙黏蛋白表达增加。与荷瘤裸鼠对照组相比，sh-CRNDE组肿瘤生长减缓，肿瘤重量减轻，E-钙黏蛋白升高，N-钙黏蛋白、波形蛋白和Snail表达降低。总之，沉默CRNDE可能抑制EMT，从而通过抑制Wnt/β-连环蛋白信号通路的激活来降低人OSCC细胞的迁移和侵袭，进而限制细胞生长并促进细胞凋亡。
