摘要
目的研究长链非编码RNA(LncRNA)核富集的转录物(NEAT)1对人口腔鳞癌细胞Tca8113增殖、凋亡和自噬的影响,并探讨其机制。方法运用qRT-PCR检测人口腔鳞癌细胞Tca8113、人正常口腔上皮细胞HOEC中NEAT1、短链非编码RNA(miR)-520b的表达;将si-NC组(转染si-NC)、si-NEAT1组(转染si-NEAT1)、miR-520b组(转染miR-520b mimics)、miR-NC组(转染miR-NC)、si-NEAT1+anti-miR-NC组(si-NEAT1和anti-miR-NC共转染)、si-NEAT1+anti-miR-520b组(si-NEAT1和anti-miR-520b共转染),用脂质体法转染至Tca8113细胞;MTT法检测各组细胞的增殖;流式细胞术检测各组细胞的凋亡;Western印迹检测各组细胞中LC3Ⅱ/Ⅰ的蛋白表达;双荧光素酶报告基因检测实验检测各组细胞的荧光活性。结果与HOEC相比,Tca8113中NEAT1表达显著升高,miR-520b表达显著降低(P<0.05);抑制NEAT1、过表达miR-520b均可抑制Tca8113细胞增殖,促进凋亡和自噬。抑制miR-520b可逆转抑制NEAT1对Tca8113细胞的增殖抑制和凋亡、自噬促进作用。结论抑制NEAT1可抑制口腔鳞癌细胞增殖、促进凋亡和自噬,其机制可能与靶向miR-520b有关,将可为口腔鳞癌的靶向治疗提供依据。
Objective To study the effects of LncRNA NEAT1 on proliferation,apoptosis and autophagy of oral squamous cell carcinoma Tca8113 and explore its mechanism.Methods qRT-PCR was used to detect the expression of NEAT1 and miR-520b in human oral squamous cell carcinoma Tca8113 and human normal oral epithelial cells HOEC;the si-NC group(transfected si-NC),si-NEAT1 group(transfected si-NEAT1),miR-520b group(transfected miR-520b mimics),miR-NC group(transfected miR-NC),si-NEAT1+anti-miR-NC group(co-transfected si-NEAT1 and anti-miR-NC),si-NEAT1+anti-miR-520b group(co-transfected si-NEAT1 and anti-miR-520b),all transfected into Tca8113 cells by liposome method.MTT assay was used to detect the proliferation of each group of cells;the apoptosis of each group was detected by flow cytometry.The expression of LC3Ⅱ/Ⅰprotein was detected by Western blot.The fluorescence activity of each group was detected by dual luciferase reporter gene assay.Results Compared with that in human normal oral epithelial cells,the expression of NEAT1 was significantly increased in human oral squamous cell carcinoma cell line Tca8113,the expression of miR-520b was significantly decreased(P<0.05).Inhibition of NEAT1 or overexpression of miR-520b inhibited proliferation,promoted apoptosis and autophagy in Tca8113 cells.NEAT1 targeted miR-520b.Inhibition of miR-520b reversed inhibition effect of NEAT1 on proliferation inhibition,apoptosis and autophagy promotion of Tca8113 cells.Conclusions Inhibition of NEAT1 could inhibit the proliferation of oral squamous cell carcinoma,promote apoptosis and autophagy and its mechanism might be related to targeting miR-520b,which will provide a basis for targeted therapy of oral squamous cell carcinoma.
作者
王斌
吕锦
孔敏
王虎
WANG Bin;L Jin;KONG Min(Department of Stomatology,Chengdu First People′s Hospital,Chengdu 610041,Sichuan,China)
出处
《中国老年学杂志》
CAS
北大核心
2019年第16期4045-4049,共5页
Chinese Journal of Gerontology
基金
国家自然科学基金(10675087)
