贵州医科大学学报

2020, v.45;No.237(06) 678-683

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miR-30a-5p对口腔鳞状细胞癌CAL-27细胞的影响及分子机制
Effect of miR-30a-5p Expression on Oral Squamous Cell Carcinoma CAL-27 Cells and its Molecular Mechanism

马明;刘滨;
MA Ming;LIU Bin;The Second Affiliated Hospital of Shenyang Medical College;

摘要(Abstract):

目的:探讨miR-30a-5p对人口腔鳞状细胞癌(OSCC) CAL-27细胞的增殖、迁移、侵袭及上皮间质转化(EMT)的影响,并探讨其分子机制。方法:人OSCC CAL-27细胞转染后,设置为miR-30a-5p mimic组(miR-30a-5p的模拟物)、NC mimic组(阴性对照模拟物)、NC inhibitor组(阴性对照抑制剂)及miR-30a-5p inhibitor组(miR-30a-5p的抑制剂),同时设未转染CAL-27细胞为Control组;采用实时荧光定量PCR检测各组细胞miR-30a-5p基因表达,采用Western blot实验检测各组细胞EMT上皮标志物E-cadherin和间质标记物N-cadherin的蛋白表达,采用3-(4,5二甲基噻唑-2)-2,5二苯基四氮唑溴盐(MTT)法、划痕实验及Transwell实验分别检测各组细胞的细胞活力、迁移能力和侵袭能力。结果:miR-30a-5p mimic组CAL-27细胞miR-30a-5p表达较NC mimic组明显上调,miR-30a-5p inhibitor组较NC inhibitor组表达明显下降(P <0. 01); miR-30a-5p mimic组CAL-27细胞OD值较NC mimic组下降,miR-30a-5p inhibitor组较NC inhibitor组上调(P <0. 05); miR-30a-5p mimic组CAL-27细胞的迁移率较NC mimic组明显下降,miR-30a-5p inhibitor组较NC inhibitor组明显上调(P <0. 01); miR-30a-5p mimic组CAL-27细胞侵袭数较NC mimic组明显下降(P <0. 01),miR-30a-5p inhibitor组较NC inhibitor组上调(P <0. 05); miR-30a-5p mimic组CAL-27细胞EMT上皮标志物E-cadherin表达较NC mimic组明显上调、间质标志物N-cadherin的表达较NC mimic组明显下调(P <0. 01),miR-30a-5p inhibitor组E-cadherin表达较NC inhibitor组明显下调、N-cadherin表达较NC inhibitor组明显上调(P <0. 01)。结论:MiR-30a-5p可抑制OSCC CAL-27细胞增殖、迁移及侵袭能力,其机制可能与miR-30a-5p抑制CAL-27细胞EMT转化有关。
Objective: To investigate the effect of miR-30a-5p on the proliferation,migration,invasion and epithelial-mesenchymal transition( EMT) of human oral squamous cell carcinoma( OSCC) CAL-27 cells and to explore its molecular mechanism. Methods: Based on transfected molecules,CAL-27 cells were divided into the following groups: miR-30a-5p mimic,negative control( NC) NC mimic,NC inhibitor,miR-30a-5p inhibitor and the untransfected CAL-27 cells as the control. Real-time quantitative PCR( qPCR) was used to detect the miR-30a-5p expression. The protein expression of EMT epithelial markers such as E-cadherin and interstitial marker N-cadherin were detected by Western blot. MTT assay was used to assess cell viability. Scratch assay and Transwell assay were used to detect cell migration and invasion,respectively. Results: Compared to the control and NC mimic,miR-30a-5p mimic upregulated the expression level of miR-30a-5p,while miR-30a-5p inhibitor downregulated the expression level of miR-30a-5p( P < 0. 01). Compared to the control and NC mimic,MTT assay showed that miR-30a-5p mimic decreased OD values while the miR-30a-5p inhibitor group increased the OD values( P < 0. 05). Likewise,scratch assay revealed that miR-30a-5p mimic significantly inhibited cell migration( P < 0. 01). Transwell assay demonstrated that miR-30a-5p mimic remarkably reduced the invasive cell numbers( P < 0. 01),while the miR-30a-5p inhibitor increased the invasive cell numbers( P < 0. 05). In contrast to miR-30a-5p inhibitor,Western blot showed that miR-30a-5p mimic significantly downregulated the expression level of Ecadherin compared to NC mimic or NC inhibitor or control( P < 0. 01). Furthermore,miR-30a-5p mimic significantly augmented the expression level of N-cadherin compared to NC mimic or NC inhibitor or control( P < 0. 01). Conclusion: miR-30a-5p inhibites the proliferation,migration and invasion of CAL-27 cells. The mechanism may be related to miR-30a-5p-inhibited EMT of CAL-27 cells.

关键词(KeyWords): 口腔鳞状细胞癌;细胞增殖;细胞迁移分析;细胞侵袭;上皮间质转化;CAL-27细胞;miR-30a-5p基因
oral sprays;cell proliferation;cell migration assays;cell invasion;epithelial-mesenchymal transition(EMT);CAL-27 cell;miR-30a-5p gene

Abstract:

Keywords:

基金项目(Foundation): 沈阳医学院科学基金项目(20181009)

作者(Author): 马明;刘滨;
MA Ming;LIU Bin;The Second Affiliated Hospital of Shenyang Medical College;

Email:

DOI: 10.19367/j.cnki.2096-8388.2020.06.011

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