中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2013年
1期
29-32
,共4页
祝葆华%陆元志%苑进凯%王坤
祝葆華%陸元誌%苑進凱%王坤
축보화%륙원지%원진개%왕곤
肝肿瘤%基因表达调控,肿瘤%肿瘤转移
肝腫瘤%基因錶達調控,腫瘤%腫瘤轉移
간종류%기인표체조공,종류%종류전이
Liver neoplasms%Gene expression regulation,neoplastic%Neoplasm metastasis
目的 探讨miR-181b对靶基因的金属蛋白酶组织抑制因子-3(tissue inhibitor of met-allo proteinases 3,TIMP-3)的调控作用及其对肝癌细胞体外侵袭迁移能力的影响.方法 采用qRT-PCR检测肝细胞癌(HCC)组织和不同人HCC细胞系中miR-181b的表达.Western-blot检测TIMP-3在HCC中蛋白的表达.选择miR-181b高表达HCC细胞系SKHep-1,通过报告基因检测试验,观察miR-181b对TIMP-3的调控作用.分别通过Transwell小室和细胞划痕试验,观察阻断SKHep-1细胞miR-181b表达对其侵袭、迁移能力的影响.结果 miR-181b在HCC组织中mRNA的表达明显高于癌旁组织和正常肝组织(P＜0.01),其高表达与HCC高侵袭转移密切相关(P＜0.01).TIMP-3在HCC中蛋白表达明显低于癌旁组织和正常肝组织(P＜0.05).miR 181b在人HCC细胞Hep3B、HepG2、Huh-7、SKHep-1、SNU182、SNU449和肝细胞中均有表达,其中在SKHep-1中表达最高(P＜0.01).高表达miR-181b抑制带有TIMP-3' UTR的荧光素酶的表达(P＜0.05),抑制miR-181b不仅能降低SK-hep1的HCC细胞迁移能力(P＜0.05),还可降低其侵袭能力(P＜0.01).结论 miR-181b可能通过下调TIMP-3基因表达促进HCC侵袭和迁移能力.
目的 探討miR-181b對靶基因的金屬蛋白酶組織抑製因子-3(tissue inhibitor of met-allo proteinases 3,TIMP-3)的調控作用及其對肝癌細胞體外侵襲遷移能力的影響.方法 採用qRT-PCR檢測肝細胞癌(HCC)組織和不同人HCC細胞繫中miR-181b的錶達.Western-blot檢測TIMP-3在HCC中蛋白的錶達.選擇miR-181b高錶達HCC細胞繫SKHep-1,通過報告基因檢測試驗,觀察miR-181b對TIMP-3的調控作用.分彆通過Transwell小室和細胞劃痕試驗,觀察阻斷SKHep-1細胞miR-181b錶達對其侵襲、遷移能力的影響.結果 miR-181b在HCC組織中mRNA的錶達明顯高于癌徬組織和正常肝組織(P＜0.01),其高錶達與HCC高侵襲轉移密切相關(P＜0.01).TIMP-3在HCC中蛋白錶達明顯低于癌徬組織和正常肝組織(P＜0.05).miR 181b在人HCC細胞Hep3B、HepG2、Huh-7、SKHep-1、SNU182、SNU449和肝細胞中均有錶達,其中在SKHep-1中錶達最高(P＜0.01).高錶達miR-181b抑製帶有TIMP-3' UTR的熒光素酶的錶達(P＜0.05),抑製miR-181b不僅能降低SK-hep1的HCC細胞遷移能力(P＜0.05),還可降低其侵襲能力(P＜0.01).結論 miR-181b可能通過下調TIMP-3基因錶達促進HCC侵襲和遷移能力.
목적 탐토miR-181b대파기인적금속단백매조직억제인자-3(tissue inhibitor of met-allo proteinases 3,TIMP-3)적조공작용급기대간암세포체외침습천이능력적영향.방법 채용qRT-PCR검측간세포암(HCC)조직화불동인HCC세포계중miR-181b적표체.Western-blot검측TIMP-3재HCC중단백적표체.선택miR-181b고표체HCC세포계SKHep-1,통과보고기인검측시험,관찰miR-181b대TIMP-3적조공작용.분별통과Transwell소실화세포화흔시험,관찰조단SKHep-1세포miR-181b표체대기침습、천이능력적영향.결과 miR-181b재HCC조직중mRNA적표체명현고우암방조직화정상간조직(P＜0.01),기고표체여HCC고침습전이밀절상관(P＜0.01).TIMP-3재HCC중단백표체명현저우암방조직화정상간조직(P＜0.05).miR 181b재인HCC세포Hep3B、HepG2、Huh-7、SKHep-1、SNU182、SNU449화간세포중균유표체,기중재SKHep-1중표체최고(P＜0.01).고표체miR-181b억제대유TIMP-3' UTR적형광소매적표체(P＜0.05),억제miR-181b불부능강저SK-hep1적HCC세포천이능력(P＜0.05),환가강저기침습능력(P＜0.01).결론 miR-181b가능통과하조TIMP-3기인표체촉진HCC침습화천이능력.
Objective To explore the impact of TIMP 3 regulated by miR-181b as a target gene on invasion and migration of hepatocellular carcinoma (HCC) in vitro.Methods The expressions of miR-181b were detected using SYBR Green real-time fluorescence quantitative polymerase chain reaction on liver cancer specimens and on HCC cell lines.The protein expression of TIMP 3 in HCC was detected using westen blot,and SKHep-1 as a cell line expressing high miR-181b was chosen through reporter gene experiment.TIMP-3 as a target gene regulated by miR-181b and its effect on invasion and migration treated by anti-miR-181 b were studied using transwell and cell scarification test,respectively.Results The expression of miR-181b in HCC was higher than cancer-adjacent tissues and normal liver tissues.The differences among them were significant.There was a correlation between the high expression of miR-181b and invasiveness and metastasis in HCC.The protein expression of TIMP-3 in HCC was significantly lower than normal liver tissues and cancer-adjacent tissues.Expression of miR-181b mRNA was detected in various HCC cell lines such as Hep3B,HepG2,Huh 7,SKHep-1,SNU182,SNU449 and hepatocyte,with the expression of miR-181b in SKHep-1 being the highest (P＜0.01).TIMP3-3UTR was low when the expression of miR-181b was high (P＜0.05).The invasion and migration abilities of SKHep-1 were significantly inhibited by anti-miR-181b (P＜0.05).Conclusion The data suggested that miR-181b promoted invasion and migration of SKHep-1 by down-regulating TIMP-3 in HCC.
