RECQ1沉默对口腔鳞癌细胞增殖、侵袭及上皮间质转化的影响

doi:10.3969/j.issn.1674-8603.2020.02.008

›› 2020, Vol. 11 ›› Issue (2): 102-106.doi: 10.3969/j.issn.1674-8603.2020.02.008

RECQ1沉默对口腔鳞癌细胞增殖、侵袭及上皮间质转化的影响

何晓南¹,刘记伟²

1. 重庆市第九人民医院
2. 重庆市第九人民医院口腔科

收稿日期:2019-04-19 修回日期:2019-12-19 出版日期:2020-06-25 发布日期:2020-07-08
通讯作者: 何晓南 E-mail:hexiaonan1985@126.com

Effects of RECQ1 Silencing on Proliferation, Invasion and Epithelial Mesenchymal Transition of Oral Squamous Cell Carcinoma

Received:2019-04-19 Revised:2019-12-19 Online:2020-06-25 Published:2020-07-08

摘要/Abstract

摘要： 目的：探讨沉默DNA解螺旋酶RECQ1基因对口腔鳞癌细胞增殖、迁移及上皮间质转化（EMT）的影响。方法：体外培养人正常口腔角质上皮HOK细胞、口腔鳞癌Cal27细胞，采用实时定量RT-PCR与Western Blot检测RECQ1 mRNA及蛋白表达水平。采用RNA干扰法靶向沉默Cal27细胞RECQ1基因，实验分为5组：si-RECQ1组（si-RECQ1转染至Cal27细胞）、si-RECQ1+转化生长因子β1（TGF-β1）组（RECQ1沉默后加入TGF-β1）、si-NC组、TGF-β1组及空白对照组。采用CCK-8法检测各组细胞增殖抑制率，Transwell法检测细胞侵袭能力，细胞划痕实验检测细胞迁移能力，采用实时定量RT-PCR与Western Blot检测RECQ1及上皮性钙黏附蛋白（E-cadherin）、N钙黏附蛋白（N-cadherin）、波形蛋白（Vimentin）等EMT相关蛋白表达。结果：与HOK细胞相比，Cal27细胞RECQ1 mRNA及蛋白表达水平均显著升高（P＜0.05）；与空白对照组、si-NC组相比，si-RECQ1组Cal27细胞中RECQ1 mRNA及蛋白表达水平均显著降低（P＜0.05）；与空白对照组、si-NC组、si-RECQ1＋TGF-β1组相比，si-RECQ1组Cal27细胞增殖抑制率、E-cadherin mRNA及蛋白表达水平均显著升高，而TGF-β1组显著降低（P＜0.05），且si-RECQ1组Cal27细胞侵袭数量、迁移距离、Vimentin、N-cadherin mRNA及蛋白表达水平均显著降低，而TGF-β1组显著升高（P＜0.05）。结论：RECQ1沉默后明显抑制口腔鳞癌细胞增殖、迁移及侵袭，其机制可能是通过抑制EMT过程而实现。

关键词: 口腔鳞癌, RECQ1, 细胞增殖, 细胞侵袭, 上皮间质转化

Abstract: Objective： To investigate the effects of silencing RECQ1 gene on the proliferation, migration, and epithelial mesenchymal transition (EMT) of oral squamous cell carcinoma. Methods： Human normal oral keratinocytes HOK cells and oral squamous cell carcinoma Cal27 cells were cultured in vitro, and the expressions of RECQ1 mRNA and protein were detected by qRT-PCR and Western Blot. RNAi was used in targeted silence RECQ1 gene in oral squamous cell carcinoma Cal27 cells. The experiment was divided into five groups: si-RECQ1 group (si-RECQ1 transfected into Cal27 cells), si-RECQ1+transforming growth factorβ1 (TGF-β1) group (added TGF-β1 after RECQ1 silence), si-NC group, TGF-β1 group and blank control group.CCK-8 method was used to detect the inhibition rate of cell proliferation in each group. Transwell assay was used to detect the invasive ability of cells. Cell migration ability was measured by cell scratch test. The expressions of RECQ1 and EMT-related proteins, including E-cadherin, N-cadherin and Vimentin, were detected by qRT-PCR and Western Blot. Results： Compared with HOK cells, the expression of RECQ1 mRNA and protein in Cal27 cells increased significantly (P＜0.05); compared with the blank control group and the si-NC group, the expressions of RECQ1 mRNA and protein in Cal27 cells in the si-RECQ1 group decreased significantly (P＜0.05); compared with blank control group, si-NC group and si-RECQ1+TGF-β1 group, the proliferation inhibition rate, E-cadherin mRNA and protein expression levels of Cal27 cells in si-RECQ1 group were increased significantly, while TGF-β1 group decreased significantly (P＜0.05), and Cal27 cell invasion number, migration distance, Vimentin, N-cadherin mRNA and protein expression levels in si-RECQ1 group decreased significantly, but TGF-β1 group increased significantly (P< 0.05). Conclusions： RECQ1 silencing can significantly inhibit the proliferation, migration and invasion of oral squamous cell carcinoma, which may be achieved by inhibiting the EMT process.

Key words: Oral squamous cell carcinoma, RECQ1, Cell proliferation, Cell invasion, Epithelial mesenchymal transition

何晓南刘记伟. RECQ1沉默对口腔鳞癌细胞增殖、侵袭及上皮间质转化的影响[J]. 口腔生物医学, 2020, 11(2): 102-106.

参考文献 19

[1]	LIU PQ, GUO ZZ, YIN YT, et al.Expression of SRSF3 is Correlated with Carcinogenesis and Progression of Oral Squamous Cell Carcinoma[J].Int J Med Sci, 2016, 13(7):533-539
[2]	Shraddha KS, Niranjan KC, Hallikeri K.Immunolocalization of c-Jun in normal mucosa,oral submucous fibrosis,epithelial dysplasia,and oral squamous cell carcinoma: A comparative study[J].J Cancer Res Ther, 2018, 14(6):1180-1183
[3]	Bill R, Christofori G.The relevance of EMT in breast cancer metastasis: Correlation or causality?[J].FEBS Lett, 2015, 589(14):1577-1587
[4]	Sun J, Wang Y, Xia Y, et al.Mutations in RECQL Gene Are Associated with Predisposition to Breast Cancer[J].PLoS Genet, 2015, 11(5):e1005228-
[5]	LI L, GAO M, SONG B, et al.Effects of RECQ1 helicase silencing on non-small cell lung cancer cells[J].Biomed Pharmacother, 2016, 83(5):1227-1232
[6]	陈慧军, 陈少豪, 陈锦添, 等.沉默Talin-1基因对前列腺癌PC-3细胞株上皮间质转化及侵袭能力的影响[J].中华老年医学杂志, 2016, 35(10):1115-1119
[7]	Routray S, Kheur S, Chougule HM, et al.Establishing Fascin over-expression as a strategic regulator of neoplastic aggression and lymph node metastasis in oral squamous cell carcinoma tumor microenvironment[J].Ann Diagn Phathol, 2017, 30(1):36-41
[8]	Ghuwalewala S, Ghatak D, Das P, et al.CD44(high)CD24(low) molecular signature determines the Cancer Stem Cell and EMT phenotype in Oral Squamous Cell Carcinoma[J].Stem Cell Res, 2016, 16(2):405-417
[9]	Lu X, Parvathaneni S, Li XL, et al.Transcriptome guided identification of novel functions of RECQ1 helicase[J].Methods, 2016, 108(1):111-117
[10]	Tao J, Tao S, Han J, et al.RECQL1 plays an important role in the development of tongue squamous cell carcinoma[J].Cell Physiol Biochem, 2014, 33(5):1579-1590
[11]	Matsushita Y, Yokoyama Y, Yoshida H, et al.The level of RECQL1 expression is a prognostic factor for epithelial ovarian cancer[J].J Ovarian Res, 2014, 7(5):107-116
[12]	Viziteu E, Klein B, Basbous J, et al.RECQ1 helicase is involved in replication stress survival and drug resistance in multiple myeloma[J].Leukemia, 2017, 31(10):2104-2113
[13]	Li XL, Lu X, Parvathaneni S, et al.Identification of RECQ1-regulated transcriptome uncovers a role of RECQ1 in regulation of cancer cell migration and invasion[J].Cell Cycle, 2014, 13(15):2431-2445
[14]	Chen Y, Huang S, Wu B, et al.Transforming growth factor-β1 promotes breast cancer metastasis by downregulating miR-196a-3p expression[J].Oncotarget, 2017, 8(30):49110-49122
[15]	Wei H, Liang F, Cheng W, et al.The mechanisms for lung cancer risk of PM25 : Induction of epithelial-mesenchymal transition and cancer stem cell properties in human non-small cell lung cancer cells[J].Environ Toxicol, 2017, 32(11):2341-2351
[16]	Parvathaneni S, Lu X, Chaudhary R, et al.RECQ1 expression is upregulated in response to DNA damage and in a p53-dependent manner[J].Oncotarget, 2017, 8(44):75924-75942
[17]	Li W, Zong S, Shi Q, et al.Hypoxia-induced vasculogenic mimicry formation in human colorectal cancer cells: Involvement of HIF-1a, Claudin-4, and E-cadherin and Vimentin[J].Sci Rep, 2016, 6:37534-
[18]	卢宏全, 钱小英, 黄国定, 等.SUMO1P3在胰腺癌组织中的表达及其沉默对胰腺癌细胞增殖、迁移、侵袭的影响[J].山东医药, 2018, 58(2):5-8
[19]	Xu CY, Qin MB, Tan L, et al.NIBP impacts on the expression of E-cadherin,CD44 and vimentin in colon cancer via the NF-κB pathway[J].Mol Med Rep, 2016, 13(6):5379-5385

RECQ1沉默对口腔鳞癌细胞增殖、侵袭及上皮间质转化的影响

Effects of RECQ1 Silencing on Proliferation, Invasion and Epithelial Mesenchymal Transition of Oral Squamous Cell Carcinoma

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