Liu Xuan, Wang Cui Xia, Feng Qin, Zhang Tao
Department of Gynecology, Women and Children's Hospital, Qingdao University, Qingdao, Shandong 266000, P.R. China.
Department of Pediatrics, Eighth People's Hospital of Qingdao Shandong, Qingdao, Shandong 266000, P.R. China.
Exp Ther Med. 2023 Sep 4;26(4):487. doi: 10.3892/etm.2023.12186. eCollection 2023 Oct.
The present study aimed to examine the effects of the long non-coding (lnc)RNA expressed by tissue differentiation-inducing non-protein coding RNA (TINCR) on cervical cancer development. For this purpose, adjacent normal and cancer tissues were obtained from patients with cervical cancer and the lncRNA TINCR level was examined using reverse transcription-quantitative PCR (RT-qPCR) and hybridization. The association between lncRNA TINCR and the clinicopathological characteristics and prognosis of patients with cervical cancer was also analyzed. Furthermore, the expression levels of lncRNA TINCR, miRNA-7, mTOR, hypoxia-inducible factor 1 subunit α and VEGF were measured using RT-qPCR and western blot analysis. Cell proliferation, apoptosis, and invasion and migration were examined using MTT assay, 5-ethynyl-2'-deoxyuridine staining, flow cytometry, TUNEL assay, and Transwell and wound healing assays. The association between lncRNA TINCR, miRNA-7 and mTOR was also examined using a luciferase assay. The results revealed that the lncRNA TINCR level was significantly increased in cervical cancer tissues and was associated with the overall survival of patients (low vs. high expression group; P=0.0391). LncRNA TINCR was also associated with the clinicopathological characteristics of patients with cervical cancer. Following the knockdown of lncRNA TINCR using small interfering (si)RNA, cell proliferation was significantly decreased and cell apoptosis was significantly increased (P<0.001 for both); cell invasion and migration were also significantly decreased (P<0.001 for both) following transfection with mimics miRNA-7. Transfection with miRNA-7 antisense oligonucleotide decreased the antitumor effects of si-TINCR in Siha and HeLa cell lines. As shown using the dual-luciferase assay, lncRNA TINCR could target miRNA-7 and miRNA-7 could directly regulate mTOR in HeLa and SiHa cell lines. In conclusion, the present study demonstrated that lncRNA TINCR could promote cervical cancer development via regulation of the miRNA-7/mTOR axis .
本研究旨在探讨组织分化诱导非蛋白质编码RNA（TINCR）表达的长链非编码（lnc）RNA对宫颈癌发展的影响。为此，从宫颈癌患者处获取相邻的正常组织和癌组织，并使用逆转录定量PCR（RT-qPCR）和杂交技术检测lncRNA TINCR水平。还分析了lncRNA TINCR与宫颈癌患者临床病理特征及预后之间的关联。此外，使用RT-qPCR和蛋白质印迹分析测量lncRNA TINCR、miRNA-7、mTOR、缺氧诱导因子1亚基α和VEGF的表达水平。使用MTT法、5-乙炔基-2'-脱氧尿苷染色、流式细胞术、TUNEL法以及Transwell和伤口愈合试验检测细胞增殖、凋亡以及侵袭和迁移情况。还使用荧光素酶试验检测lncRNA TINCR、miRNA-7和mTOR之间的关联。结果显示，lncRNA TINCR水平在宫颈癌组织中显著升高，且与患者的总生存期相关（低表达组与高表达组；P = 0.0391）。lncRNA TINCR还与宫颈癌患者的临床病理特征相关。使用小干扰（si）RNA敲低lncRNA TINCR后，细胞增殖显著降低，细胞凋亡显著增加（两者均P<0.001）；用miRNA-7模拟物转染后，细胞侵袭和迁移也显著降低（两者均P<0.001）。用miRNA-7反义寡核苷酸转染降低了Siha和HeLa细胞系中si-TINCR的抗肿瘤作用。如双荧光素酶试验所示，lncRNA TINCR可靶向miRNA-7，且miRNA-7可在HeLa和SiHa细胞系中直接调节mTOR。总之，本研究表明lncRNA TINCR可通过调节miRNA-7/mTOR轴促进宫颈癌发展。
