长链非编码RNA LINC00958在头颈部鳞状细胞癌的表达和初步研究

doi:10.13591/j.cnki.kqyx.2023.11.002

摘要/Abstract

摘要：

目的探讨LINC00958在头颈部鳞状细胞癌(head and neck squamous cell carcinoma,HNSCC)中的表达和作用。方法利用TCGA数据库分析LINC00958在HNSCC中的表达和对生存预后的影响;体外培养HNSCC细胞系CAL27和HN6,对其转染对照组和敲低组(si-LINC00958)质粒,采用实时荧光定量PCR实验检测LINC00958表达;CCK-8实验检测细胞增殖能力变化;Transwell实验检测细胞迁移和侵袭能力变化;WB实验检测转移相关蛋白E-钙黏附蛋白(E-cadherin)、N-钙黏附蛋白(N-cadherin)和波形蛋白(Vimentin)表达情况;应用TargetScan数据库预测LINC00958和miR-877-3p的结合位点,双荧光素酶报告基因实验验证;分析miR-877-3p对HNSCC细胞增殖、迁移和侵袭能力的影响。结果 LINC00958在HNSCC中高表达,并影响患者总体生存率(P<0.05)。和对照组(si-NC)相比,si-LINC00958组细胞增殖、迁移和侵袭能力降低,E-cadherin表达降低,N-cadherin和Vimentin表达升高(P<0.05);TargetScan数据库显示miR-877-3p是LINC00958的靶基因,双荧光素酶报告基因实验验证两者存在靶向关系。和对照组(NC)相比,miR-877-3p mimics组细胞增殖、迁移和侵袭能力显著降低(P<0.05)。结论 LINC00958通过靶向调控miR-877-3p促进HNSCC的增殖、迁移和侵袭。

关键词: LINC00958, miR-877-3p, 头颈部鳞状细胞癌, 增殖, 迁移

Abstract:

Objective To explore the expression and role of LncRNA LINC00958 in head and neck squamous cell carcinoma(HNSCC). Methods The expression of LINC00958 in HNSCC and its effect on survival and prognosis were analyzed using TCGA database. HNSCC cell lines CAL27 and HN6 were cultured in vitro, and the transfected control group and knockdown group(si-LINC00958)plasmid were detected by real-time quantitative PCR and western blotting(WB)assay. The cell proliferation ability was detected by CCK-8 assay. The changes of cell migration and invasion ability were detected by transwell assay. The expressions of transfer-related proteins E-cadherin, N-cadherin and vimentin were detected by WB assay. The binding sites of LINC00958 and miR-877-3p were predicted by TargetScan database, and verified by double luciferase reporter gene assay. The effects of miR-877-3p on proliferation, migration and invasion of HNSCC cells were analyzed. Results LINC00958 was highly expressed in HNSCC and affected the overall survival rate(P<0.05). Compared with the control group(si-NC), the proliferation, migration and invasion of cells in si-LINC00958 group were decreased; the expression of E-cadherin was decreased, and the expression of N-cadherin and Vimentin was increased(P<0.05). The TargetScan database showed that miR-877-3p was the target gene of LINC00958, and the targeting relationship between miR-877-3p and LINC00958 was verified by double luciferase reporter gene assay. Compared with the control group(NC), cell proliferation, migration and invasion in miR-877-3p mimics group were significantly decreased(P<0.05). Conclusion LINC00958 promotes the proliferation, migration and invasion of HNSCC by targeting miR-877-3p.

Key words: LINC00958, miR-877-3p, head and neck squamous cell carcinoma, proliferation, migration

中图分类号:

R739.85

吉幻, 李萌, 姚恩惠, 钟旖, 张雨垚, 武和明, 李斌. 长链非编码RNA LINC00958在头颈部鳞状细胞癌的表达和初步研究[J]. 口腔医学, 2023, 43(11): 968-974.

JI Huan, LI Meng, YAO Enhui, ZHONG Yi, ZHANG Yuyao, WU Heming, LI Bin. Study on the expression of LncRNA LINC00958 in head and neck squamous cell carcinoma[J]. Stomatology, 2023, 43(11): 968-974.

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引物名	序列(3'—5')
LINC00958	上游:CCATTGAAGATACCACGCTGC 下游:GGTTGTTGCCCAGGGTAGTG
E-cadherin	上游:GCCTTATGATTCTCTGCTCGTG 下游:GCCCCATTCGTTCAAGTAGTC
N-cadherin	上游:GTGAGCCTGCAGATTTTAAGGTG 下游:GTTGGCTTCAGGCTCATTTTACT
Vimentin	上游:CTGGATTCACTCCCTCTGGTT 下游:TCGTGATGCTGAGAAGTTTCGTT
GAPDH	上游:GTCAACGGATTTGGTCTGTATT 下游:AGTCTTCTGGGTGGCAGTGAAT
U6	上游:AGAGAGATTACATGGCCCCT 下游:CTAATGTCACGCACGATTCT
miR-877-3p	上游:GGGTGTCCTCTTCTCCCTC 下游:CAGTGCGTGTCGTGGAGT