长链非编码RNA SNHG16在口腔鳞癌细胞系中的表达及对其增殖和迁移的影响
| 作者: |
1顾杰林,
1李正华,
1叶翀
1 贵港市人民医院口腔科，广西 贵港 537100 |
| 通讯: |
顾杰林
Email: gujlinhs@126.com |
| DOI: | 10.3978/j.issn.2095-6959.2018.04.003 |
摘要
目的：分析长链非编码RNA (long non-coding RNA，LncRNA)SNHG16在口腔鳞癌细胞系中的表达及对细胞增殖和迁移的影响。方法：采用实时荧光定量PCR(qRT-PCR)测定口腔鳞癌细胞系Cal-27，Tca8113及舌鳞癌细胞系(tongue scale cancer cell line，TSCCA)和正常口腔上皮细胞系HOK中LncRNA SNHG16的表达，将TSCCA分成3组：SNHG16-siRNA组、NT-siRNA组及Control组。利用Lipofectamine™ 2000分别转染pcDNA3.1-LncRNA-SNHG16-siRNA，pcDNA3.1-LncRNA-SNHG16-NC及空白质粒。MTT试验和细胞划痕试验分别测定细胞增殖和迁移能力，qRT-PCR及Western印迹分别测定E2F5 mRNA和蛋白表达水平。结果：LncRNA SNHG16在口腔鳞癌细胞系Cal-27，Tca8113及TSCCA中的相对表达量高于正常口腔上皮细胞系HOK(P<0.05)。转染0，24，48，72及96 h后，SNHG16-siRNA组与NT-siRNA组的吸光度值分别为0.17±0.02 vs 0.19±0.01(P>0.05)，0.39±0.04 vs 0.40±0.05 (P>0.05)，0.53±0.05 vs 0.65±0.06(P<0.05)，0.63±0.05 vs 0.91±0.08(P<0.05)，0.85±0.08 vs 1.28±0.10(P<0.01)。SNHG16-siRNA组划痕愈合率为(27.8±2.1)%，NT-siRNA组为(56.9±5.70)%，SNHG16-siRNA组划痕愈合率低于NT-siRNA组，差异有统计学意义(P<0.01)。SNHG16-siRNA组E2F5 mRNA相对表达量为0.49±0.031，NT-siRNA组为1.0±0.04，SNHG16-siRNA组E2F5 mRNA相对表达量低于NT-siRNA组，差异有统计学意义(P<0.01)。SNHG16-siRNA组E2F5蛋白相对表达量为0.57±0.05，NT-siRNA组为1.0±0.03，SNHG16-siRNA组E2F5蛋白相对表达量低于NT-siRNA组，差异有统计学意义(P<0.05)。结论：LncRNA SNHG16在口腔鳞癌细胞系中高表达，并沉默抑制口腔磷癌细胞增殖和迁移，其机制可能与下调E2F5表达有关。
关键词：
口腔鳞癌；长链非编码RNA SNGH16；增殖；迁移
Expression of long non-coding RNA SNGH16 in oral squamous cell carcinoma and its effect on proliferation and migration
CorrespondingAuthor: GU Jielin Email: gujlinhs@126.com
DOI: 10.3978/j.issn.2095-6959.2018.04.003
Abstract
Objective: To explore the expression of long non-coding RNA (LncRNA SNGH16) in oral squamous cell carcinoma cell lines and its effect on proliferation and migration. Methods: We compared the expression of LncRNA SNHG16 in oral squamous cell carcinoma cell line, Cal-27, Tca8113 and tongue scale cancer cell line (TSCCA), and normal oral squamous cell line, HOK by quantitatie realtime PCR (qRT-PCR). The TSCCA cell line was divided into three groups: SNHG16-siRNA group transfect with pcDNA3.1-LncRNA-SNHG16-siRNA, NT-siRNA group with pcDNA3.1-LncRNA-SNHG16-NC and Control group with blank plasmid by Lipofectamine™ 2000. MTT assay and cell wound scratch assay was used to measure the proliferation and migration ability. qRT-PCR and western blot was used to measure the expression level of E2F5 mRNA and protein. Results: Compare to the normal oral squamous cell line, HOK, the expression of LncRNA SNHG16 had a higher expression in oral squamous cell carcinoma cell line, Cal-27, Tca8113 and TSCCA. After transfection of LncRNA SNGH16 for 0, 24, 48, 72 and 96 h, the proliferation of oral squamous carcinoma cells was inhibited significantly (P<0.01). The scar healing rate was (27.8±2.1)% in SNHG16-siRNA group, which was significantly lower than (56.9±5.70)% in NT-siRNA group (P<0.01). The expression level of E2F5 mRNA in SNHG16-siRNA group was 0.49±0.031, which was significantly lower than 1.0±0.04 in NT-siRNA group (P<0.01). The expression level of E2F5 protein was 0.57±0.05 in SNHG16-siRNA group, which was significantly lower than 1.0±0.03 in NT-siRNA group (P<0.05). Conclusion: LncRNA SNGH16 has a high expression in the oral squamous cancer cell lines. Silencing the expression of LncRNA SNHG16 can inhibit oral squamous carcinoma cell proliferation and migration, which mechanism may be associated with down-regulated expression of E2F5.
Keywords：
oral squamous carcinoma; long non-coding RNA SNGH16; proliferation; migration