中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2013年
5期
337-340
,共4页
王梅%宋波%王波%张军%唐建武
王梅%宋波%王波%張軍%唐建武
왕매%송파%왕파%장군%당건무
肝肿瘤%烯酯酰辅酶A水解酶1%基因表达%细胞增殖%细胞迁移%细胞侵袭%肿瘤转移
肝腫瘤%烯酯酰輔酶A水解酶1%基因錶達%細胞增殖%細胞遷移%細胞侵襲%腫瘤轉移
간종류%희지선보매A수해매1%기인표체%세포증식%세포천이%세포침습%종류전이
Liver neoplasms%Enoyl coenzyme A hydratase-1%Gene expression%Cell proliferation%Cell migration%Cell invasion%Neoplasm metastasis
目的 探讨烯酯酰辅酶A水解酶1(Ech1)过表达对小鼠低淋巴道转移潜能肝癌细胞株Hca-P体外增殖生长、迁移和侵袭能力的影响,及其Ech1表达水平与肝癌淋巴道转移潜能的关系.方法 将pcDNA3.1(+)-Ech1重组质粒和空载体pcDNA3.1(+)分别稳定转染Hca-P细胞,通过半定量逆转录聚合酶链反应和酶联免疫吸附试验检测目的基因及其蛋白表达水平；在获得过表达Ech1的PEch1细胞株和实验对照组P空载体细胞株后,采用CCK-8法检测细胞分裂增殖能力,Transwell迁移实验和Transwell侵袭实验评估Ech1过表达对Hca-P细胞迁移和侵袭能力的影响.结果 PEch1组和P空载体组细胞中Ech1 mRNA相对表达水平分别为3.21±0.43和1.44±0.03,差异有统计学意义(P=0.029).PEch1组、P空载体组和未经处理的Hca-P组(P组)细胞中Ech1蛋白表达水平分别为0.140±0.005、0.088±0.003和0.078 ±0.006,差异有统计学意义(P＜0.05).PEch1组的细胞增殖能力明显高于P空载体组和P组(P＜0.05).Transwell迁移实验显示,PEch1组、P空载体组和P组穿膜细胞数分别为(143.00±7.25)个、(95.73±3.88)个和(106.67±3.54)个,PEch1组与P空载体组和P组比较,差异均有统计学意义(均P＜0.05).Transwell侵袭实验显示,PEch1组、P空载体组和P组穿膜细胞数分别为(77.20±5.46)个、(46.34±4.35)个和(49.80±5.21)个,PEch1组与P空载体组和P组比较,差异均有统计学意义(均P ＜0.05).结论 Ech1过表达可以显著促进Hca-P细胞增殖,增强其迁移和侵袭能力.Ech1有望成为临床基因治疗肝癌的靶点之一.
目的 探討烯酯酰輔酶A水解酶1(Ech1)過錶達對小鼠低淋巴道轉移潛能肝癌細胞株Hca-P體外增殖生長、遷移和侵襲能力的影響,及其Ech1錶達水平與肝癌淋巴道轉移潛能的關繫.方法 將pcDNA3.1(+)-Ech1重組質粒和空載體pcDNA3.1(+)分彆穩定轉染Hca-P細胞,通過半定量逆轉錄聚閤酶鏈反應和酶聯免疫吸附試驗檢測目的基因及其蛋白錶達水平；在穫得過錶達Ech1的PEch1細胞株和實驗對照組P空載體細胞株後,採用CCK-8法檢測細胞分裂增殖能力,Transwell遷移實驗和Transwell侵襲實驗評估Ech1過錶達對Hca-P細胞遷移和侵襲能力的影響.結果 PEch1組和P空載體組細胞中Ech1 mRNA相對錶達水平分彆為3.21±0.43和1.44±0.03,差異有統計學意義(P=0.029).PEch1組、P空載體組和未經處理的Hca-P組(P組)細胞中Ech1蛋白錶達水平分彆為0.140±0.005、0.088±0.003和0.078 ±0.006,差異有統計學意義(P＜0.05).PEch1組的細胞增殖能力明顯高于P空載體組和P組(P＜0.05).Transwell遷移實驗顯示,PEch1組、P空載體組和P組穿膜細胞數分彆為(143.00±7.25)箇、(95.73±3.88)箇和(106.67±3.54)箇,PEch1組與P空載體組和P組比較,差異均有統計學意義(均P＜0.05).Transwell侵襲實驗顯示,PEch1組、P空載體組和P組穿膜細胞數分彆為(77.20±5.46)箇、(46.34±4.35)箇和(49.80±5.21)箇,PEch1組與P空載體組和P組比較,差異均有統計學意義(均P ＜0.05).結論 Ech1過錶達可以顯著促進Hca-P細胞增殖,增彊其遷移和侵襲能力.Ech1有望成為臨床基因治療肝癌的靶點之一.
목적 탐토희지선보매A수해매1(Ech1)과표체대소서저림파도전이잠능간암세포주Hca-P체외증식생장、천이화침습능력적영향,급기Ech1표체수평여간암림파도전이잠능적관계.방법 장pcDNA3.1(+)-Ech1중조질립화공재체pcDNA3.1(+)분별은정전염Hca-P세포,통과반정량역전록취합매련반응화매련면역흡부시험검측목적기인급기단백표체수평；재획득과표체Ech1적PEch1세포주화실험대조조P공재체세포주후,채용CCK-8법검측세포분렬증식능력,Transwell천이실험화Transwell침습실험평고Ech1과표체대Hca-P세포천이화침습능력적영향.결과 PEch1조화P공재체조세포중Ech1 mRNA상대표체수평분별위3.21±0.43화1.44±0.03,차이유통계학의의(P=0.029).PEch1조、P공재체조화미경처리적Hca-P조(P조)세포중Ech1단백표체수평분별위0.140±0.005、0.088±0.003화0.078 ±0.006,차이유통계학의의(P＜0.05).PEch1조적세포증식능력명현고우P공재체조화P조(P＜0.05).Transwell천이실험현시,PEch1조、P공재체조화P조천막세포수분별위(143.00±7.25)개、(95.73±3.88)개화(106.67±3.54)개,PEch1조여P공재체조화P조비교,차이균유통계학의의(균P＜0.05).Transwell침습실험현시,PEch1조、P공재체조화P조천막세포수분별위(77.20±5.46)개、(46.34±4.35)개화(49.80±5.21)개,PEch1조여P공재체조화P조비교,차이균유통계학의의(균P ＜0.05).결론 Ech1과표체가이현저촉진Hca-P세포증식,증강기천이화침습능력.Ech1유망성위림상기인치료간암적파점지일.
Objective To investigate the effect of enoyl coenzyme A hydratase-1 (Ech1) on the proliferation and invasion ability of mouse hepatocarcinoma Hca-P cells in vitro.Methods Recombinant pcDNA3.1 (+)-Ech1 gene and pcDNA3.1 (+) were transfected into Hca-P cells by cationic liposomes introduction.Clone of PEch1 cells that stably expressing Ech1 and clone of control Pvector cells were screened by G418.The Ech1 expression was identified subsequently by reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA),respectively.The malignant behaviors of the cell lines were compared by proliferation,invasion and migration test.Results The cell line Hca-P cells stably expressing Ech1 gene was constructed.The relative expression of Ech1 mRNA in the PEch1 group was 3.21 ± 0.43 and in the Pvector group was 1.44 ± 0.03,with a significant difference between the two groups (P =0.029).The results of ELISA revealed that the expression of Ech1 protein was 0.140 ± 0.005 in the P~h1 group,0.088 ± 0.003 in the Pvector group,and 0.078 ± 0.006 in the Hca-P group,showing a significant difference between the PEch1 group and the P and Hca-P groups (P ＜ 0.05).Transwell migration test showed that the number of penetrated cells in the PEch1 group was 143.00 ± 7.25 cells,significantly higher than that of the Pvector group (95.73 ±3.88 cells) and un-treated Hca-1 group (106.67 ±3.54 cells,both P ＜ 0.05).The Transwell invasion assay showed that the number of penetrated cells was 77.20 ± 5.46 cells in the PEch1 group,significantly higher than 46.34 ± 4.35 cells in the P group and 49.80 ± 5.21 cells in the un-treated Hca-1 group (both P ＜ 0.05).Conclusions The results showed that overexpressed Ech1 in Hca-P cells may significantly increase the cell proliferation in a time-dependent manner.The up-regulation of Ech1 may increase to some extent the migration and invasion capacity of Hca-P cells.The efforts aiming at up-regulation of Ech1 expression may become a therapeutic target in the treatment of hepatocarcinoma.
