miR-138靶向PLD2基因对口腔癌细胞增殖、迁移的影响

doi:10.19439/j.sjos.2020.01.005

摘要/Abstract

摘要： 目的：探讨miR-138靶向PLD2基因抑制口腔癌细胞增殖、迁移的机制。方法：口腔癌细胞转染miR-138后,采用RT-PCR检测细胞中miR-138的表达水平,MTT法检测细胞增殖能力,流式细胞术检测细胞转染miR-138后细胞周期的分布,Transwell迁移实验检测细胞的迁移能力;采用Western 免疫印迹实验检测胃癌细胞MMP-9、PLD2及Cyclin D1的表达水平,采用荧光素酶报告实验分析miR-138与PLD2基因的靶向关系。采用SPSS 21.0软件包对数据进行统计学分析。结果：口腔癌细胞转染miR-138后,miR-138相对表达水平为4.28±0.16,显著高于空白对照及miR-NC组（P<0.05）。荧光素酶报告基因实验发现,口腔癌细胞转染miR-138后,PLD2野生质粒荧光素酶相对活性低于其他组（P<0.05）,miR-138组的PLD2的mRNA表达水平低于空白对照及miR-NC 组（P<0.05）。口腔癌细胞转染miR-138后,miR-373组的口腔癌细胞的增殖能力低于空白对照及miR-NC组（P<0.05）。流式细胞术实验发现,miR-138组的G₀/G₁期比例为（64.39±6.49）%,显著高于空白对照组及miR-NC组（P<0.05）;miR-138组S期比例为（13.28±3.16）%,显著低于空白对照组及miR-NC组（P<0.05）;各组G₂/M期比例差异无统计学意义（P>0.05）。Transwell实验发现,口腔癌细胞转染miR-138后,迁移细胞数量为138.46±24.37,显著低于空白对照组及miR-NC组（P<0.05）。Western免疫印迹实验发现,miR-138组MMP-9的相对水平为0.14±0.04、Vimentin的相对水平为0.17±0.02、Cyclin D1的相对水平为0.15±0.03,均显著低于空白对照组及miR-NC组（P<0.05）。结论：miR-138可靶向调控PLD2基因表达,对口腔癌的增殖、迁移能力产生抑制作用。

关键词: miR-138, 口腔癌, PLD2基因, 增殖, 迁移

Abstract: PURPOSE: To investigate the effect of miR-138 targeting PLD2 gene on proliferation and migration of oral cancer cells. METHODS: After oral cancer cells were transfected with miR-138, the expression level of microRNA-138 was detected by RT-PCR assay, the proliferation ability was detected by MTT assay, and cell cycle distribution was detected by flow cytometry. Transwell migration assay was used to detect cell migration ability, Western blotting assay was used to detect the expression levels of MMP-9, PLD2 and cyclin D1 in gastric cancer cells. Luciferase assay was used to report the targeting relationship between microRNA-138 and PLD2 gene. SPSS 21.0 software package was used to analyze the data. RESULTS: After miR-138 was transfected into oral cancer cells, the relative expression level of miR-138 was 4.28±; 0.16, which was significantly higher than that of blank control and miR-NC group (P<; 0.05). Luciferase reporter gene assay showed that the relative activity of PLD2 wild plasmid luciferase was significantly lower in oral cancer cells transfected with microRNA-138 than in other groups (P<; 0.05); The expression level of PLD2 gene in miR-138 group was significantly lower than that in blank control group and miR-NC group (P<; 0.05). After oral cancer cells were transfected with miR-138, the proliferation ability of oral cancer cells in miR-373 group was significantly lower than that in control group and miR-NC group (P<; 0.05). Flow cytometry showed that the ratio of G₀/G₁ phase was (64.39±; 6.49)% in the group of miR-138, which was significantly higher than that in the blank control group and the group of miR-NC(P<; 0.05); The ratio of S phase in the group of miR-138 was(13.28±; 3.16)%, which was significantly lower than that in the blank control group and the group of miR-NC (P<; 0.05); There was no significant difference in the ratio of G₂/M phase among the groups (P>; 0.05). Transwell experiment showed that the number of migrating cells transfected with miR-138 in oral cancer cells was 138.46±; 24.37, which was significantly lower than that in blank control group and miR-NC group(P<; 0.05). Western blotting experiments showed that the relative levels of MMP-9, vimentin and cyclin D1 in the miR-138 group were 0.14±; 0.04, 0.17±; 0.02 and 0.15±; 0.03, respectively, which were significantly lower than those in the blank control group and the miR-NC group(P<; 0.05). CONCLUSIONS: miR-138 can target PLD2 gene expression and inhibit the proliferation and migration of oral cancer cells.

Key words: MiR-138, Oral cancer, PLD2 gene, Proliferation, Migration

中图分类号:

R739.8

李怀奇, 叶金海, 丁旭, 武和明. miR-138靶向PLD2基因对口腔癌细胞增殖、迁移的影响[J]. 上海口腔医学, 2020, 29(1): 25-30.

LI Huai-qi, YE Jin-hai, DING Xu, WU He-ming. Effect of mi-138 targeting PLD2 gene on proliferation and migration of oral cancer cells[J]. Shanghai Journal of Stomatology, 2020, 29(1): 25-30.

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