摘要
目的:分析基质金属蛋白酶1(MMP1)在食管鳞癌组织中的表达水平,探究miRNA-623(miR-623)靶向调控MMP1表达水平影响食管鳞癌细胞增殖、迁移、侵袭及凋亡能力的分子机制。方法:从GEO数据库中下载关于食管鳞癌微阵列数据库,分析53对食管鳞癌与癌旁组织中MMP1 mRNA表达水平。qRT-PCR及Western blot法检测人正常食管鳞状上皮细胞(HET-1A)及人食管鳞癌细胞系(TE-10、KYSE30、KYSE180)中miR-623和MMP1蛋白的表达水平。生物信息学方法分析miR-623与MMP1结合的靶向序列。双荧光素酶报告基因实验检测miR-623对MMP1的靶向调控机制。用脂质体法将miR-623 mimic转染TE-10食管鳞癌细胞系中,通过qRT-PCR及Western blot检测转染后TE-10细胞系中MMP1的表达水平。CCK-8实验观察细胞增殖能力,流式细胞术观察细胞凋亡能力,细胞划痕及Transwell侵袭实验观察细胞迁移和侵袭能力。结果:MMP1 mRNA在食管鳞癌组织中的平均表达量明显高于癌旁组织(P<0.05)。与人正常食管鳞状上皮细胞系相比,在食管鳞癌细胞系中miR-623的表达水平显著下调(P<0.05),而MMP1蛋白水平则显著上调(P<0.05)。双荧光素酶报告基因实验显示miR-623能显著影响MMP1 3'-UTR表达载体的荧光素酶活性(P<0.05)。过表达miR-623可显著降低MMP1表达水平,抑制食管鳞癌细胞系细胞的增殖、迁移和侵袭能力,促进细胞凋亡(均P<0.05)。结论:在食管鳞癌中miR-623低表达,而MMP1高表达;miR-623可靶向调控癌基因MMP1的表达,从而调控食管鳞癌细胞的增殖、凋亡、迁移和侵袭,提示其可能是一个潜在的食管鳞癌治疗靶点。
Objective:To analyze the expression level of matrix metalloproteinase 1(MMP1)in esophageal squamous cell carcinoma tissues,and explore the molecular mechanism that miRNA-623(miR-623)targets MMP1 affecting the proliferation,migration,invasion and apoptotic ability of esophageal squamous cell carcinoma cells.Methods:The microarray database on esophageal squamous cell carcinoma was downloaded from the GEO database,and 53 pairs of esophageal squamous cell carcinoma and paracancerous tissues were analyzed for the expression level of MMP1 mRNA.qRT-PCR and Western blot were used to detect the expression level of miR-623 and MMP1 protein in human normal esophageal squamous epithelial cells (HET-1A) and human esophageal squamous cell carcinoma cell lines (TE-10,KYSE30,KYSE180).Bioinformatics method was used to analyze the target sequences of miR-623 binding to MMP1.Dual luciferase reporter gene assay was performed to detect the targeting and regulatory mechanism of miR-623 on MMP1.The miR-623 mimic was transfected into TE-10 esophageal squamous cell carcinoma cell line by liposome assay,and the expression level of MMP1 in TE-10 cell line was detected by qRT-PCR and Western blot.The cell proliferation ability was observed by CCK-8 assay.The cell apoptosis ability was observed by flow cytometry,and the cell migration and invasion ability was observed by cell scratch assay and Transwell invasion assay. Results: The average expression of MMP1 mRNA in esophageal squamous cell carcinoma tissues was significantly higher than that in paracancerous tissues ( P <0.05).Compared with human normal esophageal squamous epithelial cell line,the expression level of miR-623 was significantly down-regulated in esophageal squamous cell carcinoma cell lines ( P <0.05),while the protein level of MMP1 was clearly up-regulated ( P <0.05).Dual luciferase reporter gene assay showed that miR-623 significantly affected the luciferase activity of MMP1 3'-UTR expression vector ( P <0.05).Overexpression of miR-623 significantly inhibited the MMP1 expression level,cell proliferation,migration and invasion ability of esophageal squamous carcinoma cell lines,and promoted cell apoptosis (all P <0.05). Conclusion: In esophageal squamous cell carcinoma,miR-623 was lowly expressed and MMP1 was highly expressed.miR-623 could target and regulate the expression of oncogene MMP1,thus regulating the proliferation,apoptosis,migration and invasion of esophageal squamous cell carcinoma cells,suggesting that miR-623 may be a potential therapeutic target for esophageal squamous cell carcinoma.
作者
费松
吴伟东
李丹
方明星
靳博
FEI Song;WU Weidong;LI Dan;FANG Mingxing;JIN Bo(Department of Cardio-Thoracic Surgery,Guangzhou Red Cross Hospital of Jinan University,Guangdong Guangzhou 510220,China.)
出处
《现代肿瘤医学》
CAS
2024年第14期2511-2517,共7页
Journal of Modern Oncology
基金
广东省广州市科技计划项目(编号:201902010003)。
关键词
食管鳞癌
miR-623
MMP1
增殖
凋亡
迁移
侵袭
esophageal squamous cell carcinoma
miR-623
MMP1
proliferation
apoptosis
migration
invasion
