微小RNA-155对喉鳞状细胞癌Hep2细胞侵袭能力影响的实验研究

doi:10.3760/cma.j.issn.2095-7041.2019.01.014

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摘要目的探讨微小RNA(miRNA)-155对喉鳞状细胞癌(LSCC)Hep2细胞侵袭能力的影响及其机制。方法培养LSCC Hep2细胞,分为miRNA-155组、miRNA-155 NC组,分别转染miRNA-155抑制剂(miRNA-155 inhibitor)及miRNA-155阴性对照(miRNA-155 NC)。收集两组转染48 h的LSCC Hep2细胞,采用实时荧光定量PCR(qPCR)检测miRNA-155的表达,CCK-8法检测细胞活力,Transwell小室检测细胞侵袭能力,Western blot检测细胞中基质金属蛋白酶2(MMP-2)、MMP-9、钙黏蛋白(E-cadherin)、波形蛋白(vimentin)、磷脂酰肌醇-3-羟激酶(PI3K)蛋白、蛋白激酶B(AKT)、p-AKT的表达变化。结果转染后的LSCC Hep2细胞中,miRNA-155组中miRNA-155相对表达量0.44±0.04、光密度(OD)值0.38±0.04、细胞侵袭数目(39.4±4.3)个,均明显低于miRNA-155 NC组的1.52±0.15、0.68±0.07、(157.8±12.6)个,差异均有统计学意义(t=34.082、18.229、43.531, P值均＜0.01)。miRNA-155组Hep2细胞PI3K/AKT信号通路中MMP-2、MMP-9、vimentin、PI3K、AKT、p-AKT蛋白相对表达量分别为0.38±0.03、0.25±0.02、0.29±0.02、0.28±0.03、0.99±0.05、0.24±0.02,均低于miRNA-155 NC组的1.78±0.15、1.56±0.16、1.34±0.12、1.04±0.09、1.17±0.08、0.83±0.08;而E-cadherin表达量为1.13±0.10,高于miRNA-155 NC组的0.31±0.02:两组比较,差异均有统计学意义(t=29.345、46.745、59.436、34.217、15.936、30.129、44.621, P值均＜0.01)。结论下调LSCC Hep2细胞中miRNA-155的表达,可降低Hep2细胞的活力和侵袭能力,其机制可能与调控PI3K/AKT信号通路相关蛋白的表达有关。

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	秦江波
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关键词 ：喉肿瘤, Hep2细胞, 微小核糖核酸, 侵袭能力

Abstract：Objective To investigate the effect of miRNA-155 on invasion and metastasis of laryngeal squamous cell carcinoma Hep2 cells.Methods LSCC Hep2 cells were cultured and divided into miRNA-155 group and miRNA-155 NC group, respectively, and were transfected with miRNA-155 inhibitor and miRNA-155 negative control (miRNA-155 NC). Both groups of transfected LSCC Hep2 cells were transfected for 48 h, the expression of miRNA-155 was detected by quantitative real-time fluorescence quantitative polymerase chain reaction (qPCR). Cell viability was detected by CCK-8 assay. Cell invasion was detected by transwell assay. The expressions of matrix metalloproteinase-2 (MMP-2), MMP-9, E-cadherin, Vimentin, phosphatidylinositol 3 kinase (PI3K), phosphorylation of protein kinase B (AKT) were detected by western blot.Results In the transfected Hep2 cells, the relative expression of miRNA-155 in the miRNA-155 group was 0.44 ±0.04, the optical density(OD) value was 0.38 ±0.04, and the number of cell invasion was 39.4 ±4.3, were significantly lower than those in the miRNA-155 NC group 1.52±0.15, 0.68 ±0.07, and 157.8 ±12.6. The differences were statistically significant (t=34.082, 18.229, 43.531, all P values<0.01). Compared with the expression of PI3K/AKT signaling pathway related proteins in Hep2 cells, the expression levels of MMP-2, MMP-9, Vimentin, PI3K, AKT and p-AKT in the miRNA-155 group (0.38±0.03, 0.25±0.02, 0.29±0.02, 0.28±0.03, 0.99±0.05, 0.24±0.02) were lower than those in the miRNA-155 NC group(1.78±0.15, 1.56±0.16, 1.34±0.12, 1.04±0.09, 1.17±0.08, 0.83±0.08), and the expression levels of E-cadherin(1.13±0.10) were higher than those in the miRNA-155 NC group(0.31±0.02). The differences were statistically significant(t=29.345, 46.745, 59.436, 34.217, 15.936, 30.129, 44.621, all P values＜0.01).Conclusions Down-regulating the expression of miRNA-155 in LSCC Hep2 cells can reduce cell viability and invasion, and its mechanism may be related to the regulation of the expression of PI3K/AKT signaling pathway related proteins.

Key words： Laryngeal neoplasms Hep2 cell MicroRNA Invasion ability

收稿日期: 2018-04-14

基金资助:长治医学院科技创新团队支持项目(CX201417)

通讯作者: 秦江波,Email: 1053536008@qq.com

引用本文:

秦江波, 王也煜, 常玮, 李军, 陈志飞. 微小RNA-155对喉鳞状细胞癌Hep2细胞侵袭能力影响的实验研究[J]. 中华解剖与临床杂志, 2019, 24(1): 76-81.
Qin Jiangbo, Wang Yeyu, Chang Wei, Li Jun, Chen Zhifei. Experimental study on the influence of miRNA-155 on invasion and metastasis of laryngeal squamous cell carcinoma Hep2 cells. Chinese Journal of Anatomy and Clinics, 2019, 24(1): 76-81.

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http://www.cjac.com.cn/CN/10.3760/cma.j.issn.2095-7041.2019.01.014 或 http://www.cjac.com.cn/CN/Y2019/V24/I1/76

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