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miRNAs在肿瘤中异常表达,且与肿瘤的发生发展密切相关。目前发现,miR-9-5p在肿瘤中可能发挥原癌或抑癌效应,功能尚未完全阐述清楚。本文拟探讨miR-9-5p在舌癌中的作用。前期研究中收集10例舌癌组织及配对的癌旁组织,实时荧光定量PCR技术检测后发现,miR-9-5p在舌癌组织中的表达量显著高于癌旁组织,且其在舌癌细胞中的表达量也明显高于正常舌上皮细胞。此外,在舌癌细胞Tca8113中过表达miR-9-5p显著增加细胞的增殖能力。生物信息学预测及双荧光素酶报告基因实验证实,miR-9-5p可直接结合在自噬/苄氯素1调节因子1(activating molecule in beclin1-regulated autophagy,Ambra1)的3’-UTR区域,靶向抑制Ambra1表达。Western印迹结果证实过表达miR-9-5p降低Ambra1的表达,反之亦然。Ambra1在舌癌细胞中的表达量显著低于正常舌上皮细胞。Brd U实验证实在舌癌细胞SCC-25中过表达Ambra1可显著抑制其增殖能力;相反,使用siRNA技术沉默Ambra1能够显著促进Tca8113细胞的增殖。在干预miR-9-5p的细胞中同时干预Ambra1的表达,结果发现Ambra1可显著逆转miR-9-5p对舌癌细胞增殖的促进作用。总之,miR-9-5p在舌癌中可能发挥原癌基因样作用,通过直接靶向抑制Ambra1表达进而促进舌癌细胞发生增殖。
MiRNAs are abnormally expressed in tumors and are closely related to the development of tumors. It has been found that miR-9-5p may play a proto-oncogenic or tumor suppressor effect in the tumor, the function has not yet been fully elucidated. This article intends to explore the role of miR-9-5p in tongue cancer. In the previous study, 10 cases of tongue cancer tissues and matched paracancerous tissues were collected. Real-time fluorescence quantitative PCR showed that the expression of miR-9-5p in tongue cancer tissues was significantly higher than that in paracancerous tissues, The amount of expression in the cells is also significantly higher than normal tongue epithelial cells. In addition, overexpression of miR-9-5p in tongue cancer cells Tca8113 significantly increased the cell proliferation ability. Bioinformatics prediction and dual luciferase reporter assay confirmed that miR-9-5p could directly bind to the 3’-UTR region of the autophagic / inactivation factor 1 (AMPH) -regulated autophagy , Targeting inhibition of Ambra1 expression. Western blot results confirmed that overexpression of miR-9-5p reduced Ambra1 expression and vice versa. Ambra1 expression in tongue cancer cells was significantly lower than normal tongue epithelial cells. BrdU assay confirmed that overexpression of Ambra1 in SCC-25 cells significantly inhibited the proliferation of Tca8113 cells. In contrast, silencing Ambra1 by siRNA significantly enhanced the proliferation of Tca8113 cells. In the intervention of miR-9-5p cells simultaneously interfere with the expression of Ambra1 and found that Ambra1 can significantly reverse the promotion of miR-9-5p on the proliferation of tongue cancer cells. In conclusion, miR-9-5p may play a proto-oncogene-like role in tongue cancer and promote the proliferation of tongue cancer cells by directly targeting Ambra1 expression.