摘要
目的探讨长链非编码RNA同源异形盒转录反义RNA(lncRNA HOTAIR)通过靶向微小RNA-520g-3p(miR-520g-3p)对胰腺癌细胞增殖、迁移能力的影响。方法实时定量聚合酶链式反应(qRT-PCR)检测胰腺癌细胞中miR-520g-3p、HOTAIR表达水平;转染HOTAIR小干扰RNA至SW1990胰腺癌细胞中,并分为NC组、siHOTAIR-1组、siHOTAIR-2组。细胞计数试剂盒(CCK-8)实验检测细胞增殖能力,划痕愈合实验检测细胞迁移能力;双荧光素酶报告基因实验验证miR-520g-3p与HOTAIR靶向调控关系,两组之间的定量资料比较采用t检验。结果qRT-PCR结果表明SW1990中HOTAIR mRNA水平明显高于人正常胰腺导管上皮细胞(HPDE,3.08±0.33比1.57±0.17,t=29.830,P<0.01);miR-520g-3p在SW1990中的表达水平明显低于人正常胰腺导管上皮细胞(HPDE,0.93±0.11比2.63±0.24,t=22.410,P<0.01);HOTAIR小干扰RNA转染至SW1990中,72 h后NC组细胞吸光度(A)值明显高于siHOTAIR-1组、siHOTAIR-2组(3.52±0.99比1.92±0.23、2.36±0.58,t=6.340、5.150,P<0.01);降低HOTAIR表达48 h后,NC组细胞划痕愈合面积百分比为显著高于siHOTAIR-1组、siHOTAIR-2组[(85.37±9.82)%比(50.56±5.82)%、(35.28±6.86)%,t=7.760,14.750,P<0.01],差异均有统计学意义。双荧光素酶报告基因表明miR-520g-3p可明显降低野生型HOTAIR载体荧光素酶活性。下调miR-520g-3p表达水平后,SW1990细胞增殖及迁移能力高于对照组(t=13.190、12.480,P<0.01),差异有统计学意义。结论HOTAIR可通过靶向负调控miR-520g-3p影响胰腺癌细胞增殖、迁移能力。
Objective To investigate the effects of long non-coding RNA Hox transcript antisense RNA(lncRNA HOTAIR)on proliferation and migration ability of pancreatic cancer cells by targeting microRNA-520g-3p(microRNA-520g,miR-520g-3p).Methods Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression levels of miR-520g-3p and HOTAIR in pancreatic cancer cells.HOTAIR small interfering RNA was transfected into SW1990 pancreatic cancer cells.The normal control(NC)group,siHOTAIR-1 group and siHOTAIR-2 group were set up.Cell counting kit-8(CCK-8)assay was used to detect cell proliferation ability.The scratch healing experiment was used to detect cell migration ability.The dual luciferase reporter gene experiment was used to verify the relationship between miR-520g-3p and HOTAIR targeting regulation.The quantitative data between the two groups were compared by t test.Results qRT-PCR showed that the level of HOTAIR mRNA in SW1990 was significantly higher than that in human normal pancreatic ductal epithelial cells human pancreatic duct epithelial(HPDE)(3.08±0.33 vs.1.57±0.17,t=29.830,P<0.01).The expression of miR-520g-3p in SW1990 was significantly lower than that in HPDE(0.93±0.11 vs.2.63±0.24,t=22.410,P<0.01).HOTAIR small interfering RNA was transfected into SW1990,and at72 h,A value in the NC group was significantly higher than that in siHOTAIR-1 group and siHOTAIR-2 group(3.52±0.99 vs.1.92±0.23,2.36±0.58,t=6.340,5.150,P<0.01).At 48 h after down-regulating the HOTAIR expression,the percentage of wound healing area in the NC group was significantly higher than that in the siHOTAIR-1 group and the siHOTAIR-2 group[(85.37±9.82)%vs.(50.56±5.82)%,(35.28±6.86)%,t=7.760,14.750,P<0.01].The dual luciferase reporter gene indicated that miR-520g-3p could significantly reduce the luciferase activity of wild-type HOTAIR vector.After down-regulating the expression level of miR-520g-3p,the proliferation and migration ability of SW1990 cells was significantly increased as compared with that in the control group(t=13.190,12.480,P<0.01).Conclusion HOTAIR can influence the proliferation and migration ability of pancreatic cancer cells by targeting and negatively regulating miR-520g-3p.
作者
胡明星
韩彤
余奎杨
刘传江
付强
王玉柱
秦涛
张宏伟
Hu Mingxing;Han Tong;Yu Kuiyang;Liu Chuanjiang;Fu Qiang;Wang Yuzhu;Qin Tao;Zhang Hongwei(Department of Hepatobiliary and Pancreatic Surgery,Henan Provincial People’s Hospital,Henan University,Zhengzhou 450003,China;Department of General Surgery,Henan Second Children’s Hospital,Luoyang 471000,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第1期18-20,共3页
Chinese Journal of Experimental Surgery
基金
河南省科技攻关项目(212102310151)
河南省医学科技攻关计划省部共建项目(SB201901083)。
作者简介
通信作者:张宏伟,Email:hwzhang666@126.com。
