国际外科学杂志
國際外科學雜誌
국제외과학잡지
INTERNATIONAL JOURNAL OF SURGERY
2013年
4期
238-242,封3
,共6页
刘忠考%雷长江%邱志东%李明意%黎然%包仕廷%缪辉来
劉忠攷%雷長江%邱誌東%李明意%黎然%包仕廷%繆輝來
류충고%뢰장강%구지동%리명의%려연%포사정%무휘래
mir-520c-3p%肝细胞癌%GPC3基因%细胞增生
mir-520c-3p%肝細胞癌%GPC3基因%細胞增生
mir-520c-3p%간세포암%GPC3기인%세포증생
Mir-520c-3p%Hepatocellular carcinoma%GPC3%Cell proliferation
目的 研究mir-520c-3p靶向GPC3后对肝癌Huh-7细胞增生、迁移、侵袭能力的影响,为临床寻找治疗肝癌的方法提供新的理论依据.方法 将实验分为三组:未转染mir-520c-3p的Huh-7细胞组(cel1组),阴性对照组(NC组),以及转染mir-520c-30p的Huh-7细胞组(干扰组).然后利用荧光定量PCR和Western Blotting分别检测GPC3基因的mRNA和蛋白的表达量；通过EDU检测细胞增生的变化；通过Transwell检测细胞侵袭和迁移能力的改变.结果 荧光定量PCR结果表明cell组、NC组和干扰组分别为1.13±0.23、1.28±0.15和1.05 ±0.19(P ＞0.05),mir-520c-3p不影响GPC3的mRNA转录水平；而Western Blotting实验结果表明转染mir-520c-3p后GPC3蛋白表达水平明显降低,cell组、NC组和干扰组分别为2.16 ±0.08、1.99±0.04和0.499±0.05(P＜0.01)；EDU检测发现转染mir-520c-3p后肝癌Huh-7细胞增生能力明显受抑制,cell组、NC组和干扰组分别为(90.12±1.93)％、(91.02±0.35)％和(77.73±5.88)％(P ＜0.05)；Transwell检测结果发现转mir-520c-3p后肝癌Huh-7细胞的侵袭能力受抑制,cell组、NC组和干扰组分别为0.071 ±0.008、0.105±0.001和0.048±0.002(P＜0.05)；迁移能力明显降低,其中cell组、NC组和干扰组分别为0.546 ±0.010、0.328±0.002和0.151±0.002(P ＜0.01).结论 mir-520c-3p能够靶向GPC3从而抑制肝癌Huh-7细胞的增生、侵袭、迁移能力.
目的 研究mir-520c-3p靶嚮GPC3後對肝癌Huh-7細胞增生、遷移、侵襲能力的影響,為臨床尋找治療肝癌的方法提供新的理論依據.方法 將實驗分為三組:未轉染mir-520c-3p的Huh-7細胞組(cel1組),陰性對照組(NC組),以及轉染mir-520c-30p的Huh-7細胞組(榦擾組).然後利用熒光定量PCR和Western Blotting分彆檢測GPC3基因的mRNA和蛋白的錶達量；通過EDU檢測細胞增生的變化；通過Transwell檢測細胞侵襲和遷移能力的改變.結果 熒光定量PCR結果錶明cell組、NC組和榦擾組分彆為1.13±0.23、1.28±0.15和1.05 ±0.19(P ＞0.05),mir-520c-3p不影響GPC3的mRNA轉錄水平；而Western Blotting實驗結果錶明轉染mir-520c-3p後GPC3蛋白錶達水平明顯降低,cell組、NC組和榦擾組分彆為2.16 ±0.08、1.99±0.04和0.499±0.05(P＜0.01)；EDU檢測髮現轉染mir-520c-3p後肝癌Huh-7細胞增生能力明顯受抑製,cell組、NC組和榦擾組分彆為(90.12±1.93)％、(91.02±0.35)％和(77.73±5.88)％(P ＜0.05)；Transwell檢測結果髮現轉mir-520c-3p後肝癌Huh-7細胞的侵襲能力受抑製,cell組、NC組和榦擾組分彆為0.071 ±0.008、0.105±0.001和0.048±0.002(P＜0.05)；遷移能力明顯降低,其中cell組、NC組和榦擾組分彆為0.546 ±0.010、0.328±0.002和0.151±0.002(P ＜0.01).結論 mir-520c-3p能夠靶嚮GPC3從而抑製肝癌Huh-7細胞的增生、侵襲、遷移能力.
목적 연구mir-520c-3p파향GPC3후대간암Huh-7세포증생、천이、침습능력적영향,위림상심조치료간암적방법제공신적이론의거.방법 장실험분위삼조:미전염mir-520c-3p적Huh-7세포조(cel1조),음성대조조(NC조),이급전염mir-520c-30p적Huh-7세포조(간우조).연후이용형광정량PCR화Western Blotting분별검측GPC3기인적mRNA화단백적표체량；통과EDU검측세포증생적변화；통과Transwell검측세포침습화천이능력적개변.결과 형광정량PCR결과표명cell조、NC조화간우조분별위1.13±0.23、1.28±0.15화1.05 ±0.19(P ＞0.05),mir-520c-3p불영향GPC3적mRNA전록수평；이Western Blotting실험결과표명전염mir-520c-3p후GPC3단백표체수평명현강저,cell조、NC조화간우조분별위2.16 ±0.08、1.99±0.04화0.499±0.05(P＜0.01)；EDU검측발현전염mir-520c-3p후간암Huh-7세포증생능력명현수억제,cell조、NC조화간우조분별위(90.12±1.93)％、(91.02±0.35)％화(77.73±5.88)％(P ＜0.05)；Transwell검측결과발현전mir-520c-3p후간암Huh-7세포적침습능력수억제,cell조、NC조화간우조분별위0.071 ±0.008、0.105±0.001화0.048±0.002(P＜0.05)；천이능력명현강저,기중cell조、NC조화간우조분별위0.546 ±0.010、0.328±0.002화0.151±0.002(P ＜0.01).결론 mir-520c-3p능구파향GPC3종이억제간암Huh-7세포적증생、침습、천이능력.
Objective To research the effect of mir-520c-3p targeted GPC3 to the hepatocellular carcinoma Huh-7 cell proliferation,migration,and the influence of the attack ability and find new theoretical basis for liver hepatocellular carcinoma clinical treatment.Methods The cells were divided into three groups:not transfection of mir-520c-3p group (cell group),negative control group (Nc group),and transfection of mir-520c-3p group (treat ment group).Then used fluorescence quantitative PCR and Western Blot to detect GPC3mRNA gene and protein expression quantity.Cell proliferation of change was detected by the EDU.Made use of Transwell to detect cell invasion and migration ability of the change.Results Fluorescence quantitative PCR results showed that Cell group,NC group and treatment group were 1.13 ± 0.23,1.28 ± 0.15 and 1.05 ± 0.19 (P ＞ 0.05),mir-520-3p could not reduce the GPC3mRNA; but Western Blot detection results showed that GPC3 protein expression level reduce significantly after transfection mir-520c-3p,Cell,NC and treatment group were 2.16 ± 0.08,1.99 ± 0.04 and 0.499 ± 0.05 (P ＜ 0.01).The EDU detection results showed that hepatocellular carcinoma Huh-7 cell proliferation ability obviously inhibited after transfection mir-520c-3p,Cell group,NC group and treatment group were (90.12 ± 1.93) ％,(91.02 ± 0.35) ％ and (77.73 ± 5.88) ％ (P ＜ 0.05),and Transwell test found that hepatocellular carcinoma Huh-7cell invasion abilities were restrained,Cell group,NC group and treatment group were 0.071 ±0.008,0.105 ±0.001 and 0.048 ± 0.002 (P ＜ 0.05),in the same the cells' migration abilities were reduced,Cell group,NC group and treatment group were 0.546 ± 0.010,0.328 ± 0.002 and 0.151 ± 0.002 (P ＜0.01).Conclusions Mir-520c-3p can target GPC3 so that affect hepatocellular carcinoma Huh-7 cell proliferation,invasion and migration abilities.
