Qi Xu, Yu Xiao-Jun, Wang Xu-Ming, Song Tie-Nan, Zhang Jie, Guo Xin-Zhen, Li Guo-Jun, Shao Ming
Department of Neurology, the First Affiliated Hospital of Harbin Medical University, Harbin 150001, P.R. China.
The 1st Department of Orthopedics, the First Affiliated Hospital of Harbin Medical University, Harbin 150001, P.R. China.
Mol Ther Nucleic Acids. 2019 Sep 6;17:804-818. doi: 10.1016/j.omtn.2019.06.010. Epub 2019 Jun 27.
Osteosarcoma is a malignant bone tumor, with a high incidence worldwide. The involvement of long non-coding RNAs (lncRNAs) in cancers and their molecular association with the progression of osteosarcoma have been previously discussed. We conducted the present study to examine the effect of lncRNA KCNQ1 opposite strand/antisense transcript 1 (KCNQ1OT1) on osteosarcoma cell invasion and chemosensitivity to cisplatin (CDDP). After determination of the expression of Kcnq1 in osteosarcoma tissues and cells, the plasmids with overexpression or knockdown KCNQ1OT1 were introduced into the cells to aid the identification of cell proliferation, migration, invasion, chemosensitivity to CDDP, and apoptosis. Then, the interaction between KCNQ1OT1 and the Kcnq1/DNA methyltransferase 1 (DNMT1) axis was evaluated by measuring the level of Kcnq1 promoter region methylation and DNMT1 enrichment of the Kcnq1 promoter region. Low Kcnq1 expression and high KCNQ1OT1 expression were shown in osteosarcoma tissues and cells. Kcnq1 was negatively mediated by KCNQ1OT1 via DNMT1. The overexpression of Kcnq1 or knockdown of KCNQ1OT1 inhibited the proliferation, migration, and invasion, and it promoted the chemosensitivity to CDDP and apoptosis of MG-63 cells and its CDDP-resistant cell lines. Moreover, the same trend was observed in the cells following methylation inhibitor treatment. Collectively, knockdown of KCNQ1OT1 can inhibit the osteosarcoma progression through the Kcnq1/DNMT1 axis.
骨肉瘤是一种恶性骨肿瘤，在全球范围内发病率较高。此前已有关于长链非编码RNA（lncRNA）在癌症中的作用及其与骨肉瘤进展的分子关联的讨论。我们开展了本研究，以检测lncRNA KCNQ1反义链/反义转录本1（KCNQ1OT1）对骨肉瘤细胞侵袭及对顺铂（CDDP）化疗敏感性的影响。在测定骨肉瘤组织和细胞中Kcnq1的表达后，将过表达或敲低KCNQ1OT1的质粒导入细胞，以鉴定细胞增殖、迁移、侵袭、对CDDP的化疗敏感性及凋亡情况。然后，通过检测Kcnq1启动子区域甲基化水平和Kcnq1启动子区域DNMT1富集情况，评估KCNQ1OT1与Kcnq1/DNA甲基转移酶1（DNMT1）轴之间的相互作用。骨肉瘤组织和细胞中显示出低水平的Kcnq1表达和高水平的KCNQ1OT1表达。Kcnq1通过DNMT1被KCNQ1OT1负向介导。Kcnq1的过表达或KCNQ1OT1的敲低抑制了MG-63细胞及其CDDP耐药细胞系的增殖、迁移和侵袭，并促进了其对CDDP的化疗敏感性及凋亡。此外，甲基化抑制剂处理后的细胞也观察到了相同的趋势。总体而言，敲低KCNQ1OT1可通过Kcnq1/DNMT1轴抑制骨肉瘤进展。
