长链非编码RNA HOTTIP对宫颈癌细胞增殖、迁移及侵袭的影响  

Effect of lncRNA HOTTIP on proliferation,migration and invasion of cervical cancer cells

作　　者：刘芳[1,2] 李立安[3] 张欢[2] 张唯一[3] 吴晓洁[2] 郗永义[2] 周艳荣[2] 陈红星[2] 林艳丽[2] 

机构地区：[1]安徽大学生命科学学院,合肥230601 [2]军事医学科学院生物工程研究所,北京100071 [3]解放军总医院妇产科,北京100853

出　　处：《军事医学》2015年第6期443-447,452,共6页Military Medical Sciences

摘　　要：目的研究长链非编码RNA(long non-coding RNA,lncRNA)HOXA末端转录本反义RNA(HOXA transcript at the distal tip,HOTTIP)对宫颈癌细胞增殖、迁移及侵袭能力的影响。方法将HOTTIP小干涉RNA(siRNA)及negative siRNA转染He La和C-33A宫颈癌细胞系,通过实时荧光定量PCR(q PCR)技术确认HOTTIP在细胞中表达水平的敲低,采用CCK8及克隆形成实验检测HOTTIP降低表达后对He La和C-33A细胞增殖的影响,细胞划痕实验检测HOTTIP降低表达后对He La和C-33A细胞增殖及迁移能力的影响,Matrigel细胞侵袭实验检测HOTTIP降低表达后对He La及C-33A侵袭能力的影响。结果向宫颈癌细胞系He La和C-33A中转染HOTTIP siRNA 48 h后,经q PCR检测He La及C-33A细胞中HOTTIP均出现明显下调;CCK8及克隆形成实验结果显示,HOTTIP降低表达能显著抑制He La及C-33A细胞的增殖(P<0.01);细胞划痕实验结果显示,HOTTIP降低表达能减弱He La和C-33A细胞增殖及迁移能力;Matrigel细胞侵袭实验结果显示,HOTTIP降低表达能减弱He La及C-33A侵袭能力。结论HOTTIP siRNA转染He La及C-33A宫颈癌细胞系能有效降低HOTTIP的表达,同时HOTTIP降低表达后能抑制宫颈癌细胞的增殖、迁移和侵袭能力。Objective To investigate the potential biological effect of long non-coding RNA（ lncRNA） HOXA transcript at the distal tip（ HOTTIP） on proliferation,migration and invasion of cervical cancer cells. Methods HOTTIP small interference RNA（ siRNA） was transfected into HeLa and C-33 A cervical cancer cell lines,with negative siRNA as a control. qPCR assay was performed to confirm the knock-down of the level of HOTTIP. CCK8 assay and colony-forming unit（ CFU） assay were performed to evaluate the effect of HOTTIP knock-down on HeLa and C-33 A cell proliferation. Wound healing assay was performed to evaluate the effect of HOTTIP knock-down on HeLa and C-33 A cell proliferation and migration. Tumor invasion assay was used to evaluate the effect of HOTTIP knock-down on HeLa and C-33 A cell invasion.Results The expression level of HOTTIP was efficiently knocked down by siRNA 48 h post transfection. The results of CCK8 assay and CFU assay showed that HOTTIP knock-down significantly decrease of cervical cancer cell proliferation.Wound healing assay result indicated that HOTTIP knock-down obviously suppressed cervical cancer cell proliferation and migration. Tumor invasion assay results demonstrated that HOTTIP knock-down significantly suppressed cervical cancer cell invasion. Conclusion HOTTIP levels in HeLa and C-33 A cervical cancer cell lines can be efficiently knocked down with the siRNA strategy,and the HOTTIP knock-down can significantly suppress the tumor characteristics of cervical cancer cells,including the ability of proliferation,migration and invasion.

关 键 词：lncRNA HOTTIP 宫颈肿瘤 细胞增殖 细胞运动 肿瘤浸润 

分 类 号：R737.33[医药卫生—肿瘤]