CAJ | 학술논문

目的观察肝癌细胞株MHCC97-H中E盒结合锌指蛋白Ⅰ(ZEB1)、E-钙黏蛋白(E-cadhefin)、微小RNA(miRNA,miR)-200基因的表达,探讨在肝细胞癌上皮-间充质转化中ZEB1与miR-200的关系.方法实时荧光定量聚合酶链反应(FQ-PCR)方法检测正常肝细胞株L02及肝癌细胞株MHCC97-H中ZEB1、E-cadherin及miR-200家族基因的表达量.通过短发卡RNA(shRNA)慢病毒颗粒转染L02及MHCC97-H细胞敲除ZEB1,FQ-PCR检测L02及MHCC97-H中ZEB1、E-cadherin、miR-200基因的表达量.结果 MHCC97-H组中ZEB1、miR-200a基因表达量分别为1.268 1±0.120 1、484.368 1±99.804 1,较L02组(分别为1.000 0±0.0507、220.196 8±62.021 0)均明显升高,差异有统计学意义(P＜0.05).而MHCC97-H细胞中E-cadherin基因表达量为0.443 9±0.0901,较i02组(1.0000±0.051 1)明显下降,ZEB1与E-cadherin基因表达呈负相关(r=-1.00).shRNA慢病毒颗粒成功转染L02及MHCC97-H细胞后,两组中ZEB1基因表达量分别为0.573 2±0.103 1、0.494 3±0.092 1,均明显下调(P＜0.05);两组中E-cadherin基因表达量分别为1.190 2±0.112 1、1.459 3±0.121 3,miR-200a基因表达量分别为3.0407±0.1133、2.857 1±0.113 5,miR-200b基因表达量分别为2.201 3±0.1208、3.789 1 ±0.1129,miR-200c基因表达量分别为1.665 1±0.121 7、6.129 3±0.1824,均明显上调(P＜0.05).结论在肝癌细胞株MHCC97-H中ZEB1基因表达与E-cadherin呈负相关,ZEB1与miR-200家族之间可以互相调控,从而在肿瘤细胞的增殖、侵袭转移过程中发挥重要作用.
목적 관찰간암세포주MHCC97-H중E합결합자지단백Ⅰ(ZEB1)、E-개점단백(E-cadhefin)、미소RNA(miRNA,miR)-200기인적표체,탐토재간세포암상피-간충질전화중ZEB1여miR-200적관계.방법 실시형광정량취합매련반응(FQ-PCR)방법검측정상간세포주L02급간암세포주MHCC97-H중ZEB1、E-cadherin급miR-200가족기인적표체량.통과단발잡RNA(shRNA)만병독과립전염L02급MHCC97-H세포고제ZEB1,FQ-PCR검측L02급MHCC97-H중ZEB1、E-cadherin、miR-200기인적표체량.결과 MHCC97-H조중ZEB1、miR-200a기인표체량분별위1.268 1±0.120 1、484.368 1±99.804 1,교L02조(분별위1.000 0±0.0507、220.196 8±62.021 0)균명현승고,차이유통계학의의(P＜0.05).이MHCC97-H세포중E-cadherin기인표체량위0.443 9±0.0901,교i02조(1.0000±0.051 1)명현하강,ZEB1여E-cadherin기인표체정부상관(r=-1.00).shRNA만병독과립성공전염L02급MHCC97-H세포후,량조중ZEB1기인표체량분별위0.573 2±0.103 1、0.494 3±0.092 1,균명현하조(P＜0.05);량조중E-cadherin기인표체량분별위1.190 2±0.112 1、1.459 3±0.121 3,miR-200a기인표체량분별위3.0407±0.1133、2.857 1±0.113 5,miR-200b기인표체량분별위2.201 3±0.1208、3.789 1 ±0.1129,miR-200c기인표체량분별위1.665 1±0.121 7、6.129 3±0.1824,균명현상조(P＜0.05).결론 재간암세포주MHCC97-H중ZEB1기인표체여E-cadherin정부상관,ZEB1여miR-200가족지간가이호상조공,종이재종류세포적증식、침습전이과정중발휘중요작용.
Objective To study the expression of zinfingeE-box bingdinghomeobox 1 (ZEB1) and microRN(niRNA, miR) in hepatocellulacarcinom(HCC) cell line MHCC97-H, and to furthereveal the relationship between ZEB1 and miR-200 in epithelial-mesenchymal transitionof HCC.MethodReal-time fluorescenquantitative polymerase chain reaction (FQ-PCR) waused to detecthe expression of ZEB1, E-cadherin and miR-200 in normal livecell line L-02 and MHCC97-H.The ZEB1 of MHCC97-H and L-02 cellwaknocked outhrough the shorhairpin RN(shRNA) lentiviral particletransfection of L-02 and MHCC97-H cells.FQ-PCwaused to detecthe expression of ZEB1,E-cadherin and miR-200 family in shRNL-02 and shRNA-MHCC97-H.ResultThe expression levelof ZEB1 and miR-200in MHCC97 cells-H were 1.268 1 ±0.120 1 and 484.368 1 ±99.804 1 respectively, which were significantly highethan those in L-02 cell(1.000 0 ± 0.050 7 and 220.196 8 ± 62.021 0 rcspectively) (P ＜ 0.05).The expression level of E-cadherin in MHCC97-H cellwa0.443 9 ± 0.090 1, signifycantly lowethan in L-02 cell(1.000 0 ± 0.051 1) (P ＜ 0.05).There wanegative correlation between the expression level of ZEB1 and E-cadherin in MHCC97-H cell(=-1.00).The shRNlentiviruwasuccessfully transfected into L-02 celland MHCC97-H cellrespectively.The expression level of ZEB1 in the two groupwa0.573 2 ±0.103 1, and 0.494 3 ± 0.092 1respectively, which were significantly reduced (P ＜0.05).The expression level of E-cadherin in the two groupwa1.190 2 ± 0.112 1 and 1.459 3-± 0.121 3 respectively.The expression levelof miR-200a, miR-200and miR-200in L-02 cellwere 3.040 7 ±0.113 3,2.201 3 ±0.120 8 and 1.665 1 ±0.121 7 respectively, and those in MHCC97-H cellwere 2.857 1 ± 0.113 5,3.789 1 ± 0.112 9 and 6.129 3 ± 0.182 4 respectively.They were all significantly increased (P ＜ 0.05).Conclusion The expression level of ZEB1 in MHCC97-H cellwanegatively correlated with thaof E-cadherin.They are regulated each othebetween ZEB1 and miR-200 family in MHCC97-H cells,which may play an importanrole in the proliferation, invasion and metastasiprocesof tumocells.