临床与病理杂志2018，Vol.38Issue(1)：5-11,7.DOI:10.3978/j.issn.2095-6959.2018.01.002
长链非编码RNA ANRIL对口腔鳞状细胞癌增殖和凋亡的影响及其机制
Effect of long non-coding RNA ANRIL on proliferation and apoptosis in oral squamous cell carcinoma and its mechanism
摘要
Abstract
Objective: To investigate the influence of long non-coding RNA (IncRNA) ANRIL on proliferation and apoptosis in oral squamous cell carcinoma cell line and its mechanism. Methods: The real time fluorescence quantitative PCR was used to detect and compare IncRNA ANRIL level in normal squamous cell line, HOK and four kinds of human oral squamous cell carcinoma cell line(SCC-4, Cal-27; TSCCA and Tca8113). The TSCCA cell line was divided into two groups, ANRIL gene silencing group (transfected with IncRNA-ANRILshRNA by Lipofetamine(TM) 2000) and control group (transfected with negative randomized LncRNA-ANRILNC sequence by Lipofetamine(TM) 2000). The proliferation capability and colony forming efficiency was detected by CCK-8 kit and colony formation assay respectively. The cell apoptosis rate was detected and compared upon FACS method between two groups. Finally, the protein levels of Cleaved Caspase-3, KLF2 and P21 was measured and compared between two groups by western blot. Results: The expression level of LncRNA ANRIL among four kinds of human oral squamous cell carcinoma cell line (SCC-4, Cal-27, TSCCA and Tca8113) was significantly higher than that in normal squamous cell line, HOK (P＜0.05). After transfection for 0, 24, 48, 72 and 96 hours, the OD 450 nm value of control group vs ANRIL gene silencing group was 0.38±0.04 vs 0.37±0.04(P＞0.05), 0.58±0.04 vs 0.51±0.05(P＞0.05), 0.89±0.1 vs 0.67±0.06 (P＞0.05), 1.20±0.09 vs 0.83±0.07(P＜0.05) and 1.87±0.13 vs 1.16±0.10(P＜0.Ol), respectively. The colony formation rate was 21.54％±2.03％ and 53.72％±5.93％ in ANRIL gene silencing group and control group, respectively (P＜0.01). The result of FACS method demonstrated that the cell apoptosis rate was 14.7％±0.9％ in the ANRIL gene silencing group and 4.9％±0.7％ in the control group, respectively (P＜0.01). Western blot indicated that the expression level of cleaved caspase-3 protein was 4.1±0.44 in ANRIL gene silencing group, which was significantly higher than that in control group (P＜0.01). The expression level of KLF2 protein was 0.53±0.06 in ANRIL gene silencing group, which was significantly lower than 1.0±0.03 in the control group (P＜0.05). While the expression level of P21 was 0.46±0.05 in the ANRIL gene silencing group, which was significantly lower than 1.0±0.03 in the control group (P＜0.05). Conclusion: LncRNA ANTIL was over-expression in human oral squamous cell carcinoma cell line. Silencing IncRNA ANTIL could inhibit proliferation and induce apoptosis in human oral squamous cell carcinoma; which mechanism may be associated with up-regulation of Caspase-3, down-regulation of KLF2 and P21 protein.关键词
长链非编码RNA/口腔鳞状细胞癌/增殖/凋亡Key words
IncRNA ANRIL/oral squamous cell carcinoma/proliferation/apoptosis引用本文复制引用
杨艳飞,邢育珍..长链非编码RNA ANRIL对口腔鳞状细胞癌增殖和凋亡的影响及其机制[J].临床与病理杂志,2018,38(1):5-11,7.基金项目
湖北省自然科学基金(2016CFB670). This work was supported by Natural Science Foundation of Hubei Province, China,(2016CFB670). （2016CFB670）
