LINC02154和miR-335对喉鳞癌细胞增殖和凋亡的影响-现代医学

LINC02154和miR-335对喉鳞癌细胞增殖和凋亡的影响

单位：1. 榆林市第一医院绥德院区口腔科, 陕西榆林 718000;
2. 榆林市口腔医院颌面外科, 陕西榆林 719000

分类号：R739.65

出版年·卷·期（页码）：2021·49·第七期（781-788）

摘要：

目的：研究长链非编码RNA （lncRNA） LINC02154和微小RNA-335（miR-335）对喉鳞癌细胞增殖和凋亡的影响。方法：实时定量聚合酶链反应（qRT-PCR）检测LINC02154和miR-335在喉鳞癌及邻近正常黏膜组织中的表达。在喉鳞癌细胞FD-LSC-1中转染LINC02154小干扰RNA si-LINC02154或miR-335模拟物，或共转染si-LINC02154和miR-335抑制剂。采用细胞计数试剂盒8（CCK-8）检测细胞增殖，克隆形成实验测定细胞克隆形成，流式细胞术评估细胞凋亡，蛋白质印迹分析（Western blotting）测定细胞核相关抗原Ki-67（Ki-67）、活化的天冬氨酸特异性半胱氨酸蛋白酶3（Cleaved-caspase3）、Pro-caspase3蛋白表达。生物信息学预测和双荧光素酶报告基因检测分析LINC02154与miR-335的靶向关系。结果：与邻近正常黏膜组织相比，喉鳞癌组织中的LINC02154表达水平升高，而miR-335表达水平降低，差异有统计学意义（P<0.001）。干扰LINC02154降低喉鳞癌细胞FD-LSC-1的存活率、克隆形成数、Ki67、Pro-caspase3蛋白的表达水平，并提高细胞的凋亡率、Cleaved-caspase3蛋白的表达水平，差异均有统计学意义（均P<0.001），与miR-335过表达时的结果相同。干扰miR-335后，干扰LINC02154对喉鳞癌细胞FD-LSC-1存活、克隆形成、Ki-67、Pro-caspase3蛋白表达的抑制作用和对细胞凋亡、Cleaved-caspase3蛋白表达的促进作用被逆转。LINC02154可以靶向、负调控miR-335的表达。结论：LINC02154通过靶向喉鳞癌细胞中的miR-335，调控喉鳞癌细胞增殖和凋亡过程。

Objective: To study the effect of long non-coding RNA (lncRNA) LINC02154 and microRNA (miR)-335 on the proliferation and apoptosis of laryngeal squamous carcinoma cells. Methods: Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of LINC02154 and miR-335 in laryngeal squamouscarcinoma and adjacent normal mucosa tissues. Transfected LINC02154 small interfering RNA si-LINC02154 or miR-335 mimics, or co-transfected si-LINC02154 and miR-335 inhibitors into laryngeal squamous carcinoma cell line FD-LSC-1. Cell proliferation was detected by Cell Counting Kit 8 (CCK-8), clone formation experiment was applied to determine cell clone formation, flow cytometry evaluated apoptosis, and Western blotting analysis was performed to determine nuclear-associated antigen Ki-67 (Ki-67), Cleaved-caspase3, Pro-caspase3 protein expression. Bioinformatics prediction and dual luciferase reporter gene detection analyzed the targeting relationship between LINC02154 and miR-335.Results: Compared with adjacent normal mucosa tissues, the expression level of LINC02154 in laryngeal squamous carcinoma tissue increased, while the expression level of miR-335 decreased, the difference was statistically significant (P<0.001). Interfering with LINC02154 reduced the survival rate, the number of clone formation, the expression level of Ki-67, Pro-caspase3 protein of laryngeal squamous cell carcinoma FD-LSC-1, increased the apoptosis rate of the cell and the expression level of Cleared-caspase3 protein, and the differences were all statistically significant (all P<0.001), which was the same as the results when miR-335 was overexpressed. After interfering with miR-335, the inhibitory effect of LINC02154 on the survival, clonal formation, Ki-67, Pro-caspase3 protein expression of laryngeal squamous carcinoma cell line FD-LSC-1 and the promotion of apoptosis, Cleaved-caspase3 protein expression were reversed. LINC02154 targeted and down-regulated the expression of miR-335.Conclusion: LINC02154 regulates the proliferation and apoptosis of laryngeal squamous carcinoma cells by targeting miR-335 in laryngeal squamous carcinoma cells.

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