CAJ | 학술논문

目的观察微小RNA(miRNA,miR)-221在正常肝细胞和肝癌细胞株中的表达,检测miR-221在正常肝组织和肝癌组织中的表达,并干预肝癌细胞中miR-221水平探讨其对第10号染色体缺失的磷酸酶及张力蛋白同源物基因(PTEN)的调控和对肝癌细胞生物学特性的影响.方法采用实时定量聚合酶链反应法检测miR-221在正常肝细胞和肝癌细胞株中的表达,收集28例伴有门静脉癌栓的肝细胞癌手术患者正常肝组织、癌栓、癌组织和癌旁组织并检测其表达;采用miR-221抑制剂(inhibitor)和拟似物(mimic)靶向干预miR-221在肝癌细胞中的表达,Western blot法检测其对PTEN的调控,以及细胞计数试剂盒(CCK-8)法、流式细胞仪和Transwell法检测其对肝癌细胞的增殖、凋亡和迁移的影响.结果肝癌HepG2、BEL-7402和Hep3B细胞的miR-221相对表达量分别为0.783±0.021、0.954 ±0.021、0.837±0.026,显著高于正常肝L02细胞的0.081±0.007(P ＜0.05),且肝癌组织和癌栓的相对表达量为0.897±0.048、0.984±0.048,亦显著高于正常肝组织和癌旁组织的0.067±0.007、0.063 ±0.008(P ＜0.05),而肝癌细胞的PTEN的表达水平显著低于正常肝细胞(P＜0.05),且肝癌组织和癌栓亦低于正常肝细胞(P＜0.05);与对照组比较,miR-221 inhibitor和mimic转染肝癌细胞后,PTEN表达水平分别上调和下调263％和32％ (P＜0.05),转染miR-221 inhibitor肝癌细胞的增殖和迁移活性降低,凋亡率增加(P＜0.05),而转染miR-221 mimic肝癌细胞则增殖和迁移活性增加,凋亡率降低(P＜0.05).结论肝癌细胞和肝癌组织、癌栓中miR-221呈高表达,降低其水平能上调PTEN的表达,有效降低细胞增殖和迁移活性,诱导细胞凋亡,是肝细胞癌的潜在治疗靶点.
목적 관찰미소RNA(miRNA,miR)-221재정상간세포화간암세포주중적표체,검측miR-221재정상간조직화간암조직중적표체,병간예간암세포중miR-221수평탐토기대제10호염색체결실적린산매급장력단백동원물기인(PTEN)적조공화대간암세포생물학특성적영향.방법 채용실시정량취합매련반응법검측miR-221재정상간세포화간암세포주중적표체,수집28례반유문정맥암전적간세포암수술환자정상간조직、암전、암조직화암방조직병검측기표체;채용miR-221억제제(inhibitor)화의사물(mimic)파향간예miR-221재간암세포중적표체,Western blot법검측기대PTEN적조공,이급세포계수시제합(CCK-8)법、류식세포의화Transwell법검측기대간암세포적증식、조망화천이적영향.결과 간암HepG2、BEL-7402화Hep3B세포적miR-221상대표체량분별위0.783±0.021、0.954 ±0.021、0.837±0.026,현저고우정상간L02세포적0.081±0.007(P ＜0.05),차간암조직화암전적상대표체량위0.897±0.048、0.984±0.048,역현저고우정상간조직화암방조직적0.067±0.007、0.063 ±0.008(P ＜0.05),이간암세포적PTEN적표체수평현저저우정상간세포(P＜0.05),차간암조직화암전역저우정상간세포(P＜0.05);여대조조비교,miR-221 inhibitor화mimic전염간암세포후,PTEN표체수평분별상조화하조263％화32％ (P＜0.05),전염miR-221 inhibitor간암세포적증식화천이활성강저,조망솔증가(P＜0.05),이전염miR-221 mimic간암세포칙증식화천이활성증가,조망솔강저(P＜0.05).결론 간암세포화간암조직、암전중miR-221정고표체,강저기수평능상조PTEN적표체,유효강저세포증식화천이활성,유도세포조망,시간세포암적잠재치료파점.
Objective To investigate the expression level of microRN(miRNA, miR)-221 in cell line(normal liveand hepatomcarcinomcells) and tissue(normal liveand hepatomcarcinomtissues), and to explore the regulation to phosphatase and tensin homolog deleted on chromosome ten (PTEN) and iteffecon the biological behaviothrough interfering miR-221 in hepatomcarcinomcells.MethodThe expression level of miR-221 waexamined in normal liveand hepatomcarcinomcells, in normal livetissue, tumoembolus, tumoand adjacennoncanceroutissuefrom 28 caseof hepatocellulacarcinomwith portal vein tumothrombuby real-time quantitative polymerase chain reaction.The level of miR-221 in hepatomcarcinomcellwatargetedly interfered by miR-221 inhibitoand mimic, and then itregulation to PTEN wainvestigated by Western blotting, iteffecon biological behaviorincluding cell proliferation, apoptosiand migration were examined by cell counting kit-8 (CCK-8) assay, flow cytometry and transwell assay, respectively.ResultThe relative expression level of miR-221 in hepatomcarcinomHepG2, BEL-7402 and Hep3cellwa0.783 ± 0.021, 0.954 ± 0.021 and 0.837 ± 0.026, which were significantly highethan 0.081-± 0.007 in normal liveL02 cell(P ＜ 0.05),and miR-221 in livetumoand adjacennoncanceroutissuewa0.897 ± 0.048 and 0.984 ± 0.048,which were significantly highethan 0.067 ± 0.007 and 0.063 ± 0.008 in normal liveand adjacennoncanceroutissues, respectively (P ＜ 0.05).When miR-221 inhibitoand mimiwere transfected into HepG2 cells, compared to control cells, the expression level of PTEN were 263％ and 32％, respectively (P ＜ 0.05), the cell proliferation and migration rate were decreased and the apoptosirate waincreased in HepG2 celltransfected with miR-221 inhibito(P ＜ 0.05), whereathe cell proliferation and migration rate were increased and the apoptosirate wadecreased in HepG2 celltransfected with miR-221 mimi(P＜0.05).Conclusion The level of miR-221 in hepatomcarcinomcells, livetumoand tumoembolutissuewahigh.Down-regulation of miR-221 could up-regulate the expression of PTEN, availably decrease the cell proliferation and migration and induce cell apoptosis, which mighbe potential therapitargeof hepatocellulacarcinoma.