天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (9): 902-906.

• 细胞与分子生物学 • 上一篇    下一篇

反义抑制 miRNA-155 对皮肤鳞状细胞癌 A431 细胞殖、凋亡及侵袭力的影响

时磊 1，魏明 2   

  1. 1 郑州大学第五附属医院药学部（邮编 450052），2 检验科
  • 收稿日期:2017-04-12 修回日期:2017-06-19 出版日期:2017-09-15 发布日期:2017-09-25
  • 通讯作者: 魏明 E-mail:gushiweiming@126.com

The influence of silencing miRNA-155 on proliferation and apoptosis of human cutaneous squamous cell carcinoma cell line A431

SHI Lei1, WEI Ming2   

  • Received:2017-04-12 Revised:2017-06-19 Published:2017-09-15 Online:2017-09-25
  • Contact: wei ming E-mail:gushiweiming@126.com

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摘要： 摘要：目的 观察转染反义微小核糖核酸（miRNA）-155 对人皮肤鳞状细胞癌 A431 细胞增殖、凋亡、迁移和侵 袭力的影响。方法 采用脂质体介导对皮肤鳞状细胞癌 A431 细胞转染 miRNA-155 无义序列（阴性对照组）、转染 反义 miRNA-155（转染组），空白对照组细胞不作处理。实时定量聚合酶链式反应（qRT-PCR）检测转染后 A431 细 胞 miRNA-155 的相对表达水平；采用四甲基偶氨唑盐（MTT）检测转染后皮肤鳞状细胞癌 A431 细胞增殖活性；采用 流式细胞术（FCM）检测转染后皮肤鳞状细胞癌 A431 细胞周期改变和凋亡变化；采用 Transwell 法检测侵袭力与细胞 迁移能力。结果 转染组 A431 细胞中 miRNA-155 mRNA 的相对表达水平低于空白对照组和阴性对照组（F= 634.57，P＜0.01），但空白对照组和阴性对照组比较差异无统计学意义。转染 72 h 后，转染组较空白对照组和阴性对 照组细胞存活率明显降低，转染 120 h 降低最为明显（P＜0.05）。转染组 G0/G1期细胞增多，S 期细胞减少，整体细胞 增殖指数降低，A431 细胞凋亡率、细胞迁移和侵袭力升高（P＜0.05），但各组 G2/M 期细胞周期变化、后 2 组 A431 细 胞凋亡率、细胞迁移和侵袭力差异无统计学意义。结论 皮肤鳞状细胞癌 A431 细胞转染反义 miRNA-155 可减少 miRNA-155 的表达，有效抑制 A431 细胞增殖，促进其凋亡。miRNA-155 可能成为治疗皮肤鳞状细胞癌基因表达调 控的新靶点。

关键词: 皮肤鳞状细胞癌, 微小核糖核酸-155, 反义寡核糖核酸, 增殖, 凋亡

Abstract: Abstract：Objective To investigate the effects of antisense oligonucleotide against miRNA-155 (AS-miRNA-155) on proliferation，apoptosis and invasion and migration abilities of human cutaneous squamous cell carcinoma cell line A431. Methods AS- miRNA- 155 was transfected into human cutaneous squamous cell carcinoma A431 cells by using LipofectamineTM 2000. Blank control without transfection and transfected with non-sense sequence were used as non-sense sequence control. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the expression of miRNA-155 in A431 cells. Cell proliferation was analyzed using dimethyl thiazolyldiphenyl tetrazolium (MTT) assay. Cell cycle arrest and apoptosis were studied by flow cytometry (FCM). Invasion and migration were measured by Transwell chamber assays. Results The relative expression of miRNA-155 mRNA was lower in the transfection group than that in the blank control group and the negative control group (F=634.57, P＜0.01), but there was no significant difference between the blank control group and the negative control group. After 72 h transfection, the survival rate was significantly lower in the transfection group than that of the blank control group and the negative control group, and the transfection rate decreased significantly by 120 h (P＜0.05). Cells of G0/G1 phase increased, Cells of S phase reduced, the overall PI value decreased in transfection group, and the apoptosis rate of A431 cells, migration and invasion of cells increased (P＜0.05). There was no significant difference in G2/M cycle between transfection group, blank control group and negative control group. There were no significant differences in A431 cell apoptosis rate, cell migration and invasive ability between blank control group and negative control group. Conclusion Antisense oligonucleotide against miRNA-155 can inhibit the expression of miRNA- 155, the proliferation and promote the apoptosis of human cutaneous squamous cell carcinoma A431 cells, which indicates that miRNA-155 may become a new target for the regulation of gene expression in cutaneous squamous cell carcinoma.

Key words: Cutaneous squamous cell carcinoma, MicroRNA-155, Antisense oligonucleotide, Proliferation, Apoptosis