miR-199a-5p 抑制人喉癌细胞的增殖和侵袭。
MiR-199a-5p suppresses proliferation and invasion of human laryngeal cancer cells.
机构信息
Department of Otorhinolaryngology Head and Neck Surgery, the First Hospital of Jilin University, Changchun, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Dec;24(23):12200-12207. doi: 10.26355/eurrev_202012_24010.
OBJECTIVE
To explore the roles of micro ribonucleic acid (miR)-199a-5p in the proliferation, apoptosis, invasion and metastasis of laryngeal cancer cells, and its molecular mechanisms.
PATIENTS AND METHODS
The expression of miR-199a-5p in 25 cases of laryngeal cancer tissues and paracancerous tissues was detected via quantitative real-time polymerase chain reaction (qRT-PCR). Its expression in TU212, TU686 and human epithelial type 2 (HEp-2) laryngeal cancer cell lines and normal nasopharyngeal epithelial cell line NP69 was also detected via qRT-PCR. HEp-2 cells were transiently transfected with miR-199a-5p mimic or miR-199a-5p inhibitor, and the expression of miR-199a-5p was verified using RT-PCR after transfection. The regulatory effects of miR-199a-5p on the proliferation, apoptosis, invasion and migration abilities of HEp-2 cells were observed through methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, wound healing assay and transwell assay, respectively. Then, the mechanisms of miR-199a-5p in regulating Caspase-3 activity and epithelial-mesenchymal transition (EMT)-related proteins were further explored.
RESULTS
The qRT-PCR results revealed that miR-199a-5p was significantly lowly expressed in the laryngeal cancer tissues and tumor cell lines, and overexpression of miR-199a-5p substantially inhibited the proliferation of HEp-2 cells. According to the results of flow cytometry, overexpression of miR-199a-5p promoted the apoptosis of HEp-2 cells, whereas down-regulating miR-199a-5p suppressed their apoptosis. It was found that the activity of Caspase-3 was notably enhanced after overexpression of miR-199a-5p, which was evidently weakened after down-regulating miR-199a-5p. Wound healing assay and transwell assay results manifested that overexpressing miR-199a-5p weakened the invasion and migration abilities of HEp-2 cells, which were facilitated by down-regulating miR-199a-5p. Based on Western blotting results, miR-199a-5p regulated the expressions of E-cadherin, N-cadherin and vimentin. Overexpression of miR-199a-5p could inhibit EMT process, whereas suppressing miR-199a-5p could accelerate the process.
CONCLUSIONS
The expression of miR-199a-5p in laryngeal cancer tissues is substantially lower than that in the paracancerous tissues. MiR-199a-5p suppresses proliferation, invasion and migration in laryngeal cancer cell proliferation, while triggers cell apoptosis.
目的
探讨微小 RNA（miR）-199a-5p 在喉癌细胞增殖、凋亡、侵袭和转移中的作用及其分子机制。
患者和方法
通过实时定量聚合酶链反应（qRT-PCR）检测 25 例喉癌组织和癌旁组织中 miR-199a-5p 的表达。通过 qRT-PCR 检测 TU212、TU686 和人上皮型 2（HEp-2）喉癌细胞系和正常鼻咽上皮细胞系 NP69 中 miR-199a-5p 的表达。用 miR-199a-5p 模拟物或 miR-199a-5p 抑制剂瞬时转染 HEp-2 细胞，转染后通过 RT-PCR 验证 miR-199a-5p 的表达。通过甲基噻唑基四唑（MTT）测定、流式细胞术、划痕愈合试验和 Transwell 试验分别观察 miR-199a-5p 对 HEp-2 细胞增殖、凋亡、侵袭和迁移能力的调节作用。然后，进一步探讨了 miR-199a-5p 调节 Caspase-3 活性和上皮-间充质转化（EMT）相关蛋白的机制。
结果
qRT-PCR 结果显示，miR-199a-5p 在喉癌组织和肿瘤细胞系中表达明显下调，过表达 miR-199a-5p 可显著抑制 HEp-2 细胞的增殖。根据流式细胞术的结果，过表达 miR-199a-5p 促进了 HEp-2 细胞的凋亡，而下调 miR-199a-5p 则抑制了其凋亡。结果发现，过表达 miR-199a-5p 后 Caspase-3 的活性明显增强，下调 miR-199a-5p 后则明显减弱。划痕愈合试验和 Transwell 试验结果表明，过表达 miR-199a-5p 可减弱 HEp-2 细胞的侵袭和迁移能力，下调 miR-199a-5p 则可促进其侵袭和迁移能力。Western blot 结果表明，miR-199a-5p 调节 E-钙粘蛋白、N-钙粘蛋白和波形蛋白的表达。过表达 miR-199a-5p 可抑制 EMT 过程，而抑制 miR-199a-5p 则可加速 EMT 过程。
结论
喉癌组织中 miR-199a-5p 的表达明显低于癌旁组织。miR-199a-5p 抑制喉癌细胞增殖、侵袭和迁移，诱导细胞凋亡。
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