CAJ | 학술논문

目的：探讨环氧合酶‐2（COX‐2）在稳定表达乙型肝炎病毒X蛋白（HBx）的HL7702肝细胞增殖中的作用及机制。方法 CCK‐8法及克隆形成实验检测HL7702／HBx细胞、HL7702／Mock细胞、HL7702细胞增殖情况，RT‐PCR法检测上述3种细胞中 COX‐2 mRNA 表达水平，Western‐blot 法检测 COX‐2蛋白表达水平， COX‐2选择性抑制剂NS‐398作用于各组细胞后再检测以上各组细胞增殖情况及COX‐2蛋白表达水平的改变。结果细胞增殖实验及克隆形成实验提示，HL7702／HBx 组细胞增殖能力强于 HL7702／Mock和 HL7702组（P＜0．05），克隆形成率也更高（P＜0．05）。RT‐PCR检测结果提示，相对于 HL7702／Mock细胞和 HL7702组细胞，HL7702／HBx组细胞内COX‐2的mRNA相对表达量明显增高（P＜0．05）。Western‐blot检测提示，HL7702／HBx组细胞中的COX‐2蛋白相对表达水平较高（P＜0．05），而 HL7702／Mock及 HL7702组细胞之间则无明显差别。NS‐398以时间依赖的方式部分抑制各组细胞的增殖能力，对 HL7702／HBx组细胞增殖的抑制强于其他2组细胞（P＜0．05）。经50μmol／L NS‐398处理后，3组细胞的COX‐2蛋白水平均显著降低，此现象在 HL7702／HBx组细胞中更加明显（P＜0．05）。结论 HBx蛋白可以上调肝细胞COX‐2表达，促进肝细胞增殖，选择性COX‐2抑制剂可部分逆转该作用。
목적：탐토배양합매‐2（COX‐2）재은정표체을형간염병독X단백（HBx）적HL7702간세포증식중적작용급궤제。방법 CCK‐8법급극륭형성실험검측HL7702／HBx세포、HL7702／Mock세포、HL7702세포증식정황，RT‐PCR법검측상술3충세포중 COX‐2 mRNA 표체수평，Western‐blot 법검측 COX‐2단백표체수평， COX‐2선택성억제제NS‐398작용우각조세포후재검측이상각조세포증식정황급COX‐2단백표체수평적개변。결과세포증식실험급극륭형성실험제시，HL7702／HBx 조세포증식능력강우 HL7702／Mock화 HL7702조（P＜0．05），극륭형성솔야경고（P＜0．05）。RT‐PCR검측결과제시，상대우 HL7702／Mock세포화 HL7702조세포，HL7702／HBx조세포내COX‐2적mRNA상대표체량명현증고（P＜0．05）。Western‐blot검측제시，HL7702／HBx조세포중적COX‐2단백상대표체수평교고（P＜0．05），이 HL7702／Mock급 HL7702조세포지간칙무명현차별。NS‐398이시간의뢰적방식부분억제각조세포적증식능력，대 HL7702／HBx조세포증식적억제강우기타2조세포（P＜0．05）。경50μmol／L NS‐398처리후，3조세포적COX‐2단백수평균현저강저，차현상재 HL7702／HBx조세포중경가명현（P＜0．05）。결론 HBx단백가이상조간세포COX‐2표체，촉진간세포증식，선택성COX‐2억제제가부분역전해작용。
Objective To investigate the effect of cyclooxygenase‐2(COX‐2) on the cell prolifera‐tion in HL7702 cells w hich express hepatitis B virus X protein (HBx ) stably . Methods Cell‐counting Kit‐8 (CCK‐8) assay and clone formation assay were explored to detect the proliferation of HL 7702/HBx , HL7702/Mock and HL7702 cells ,RT‐PCR and Western‐bolt were used to examine the level of COX‐2 mRNA and protein expression in cells of three groups . The level of cell proliferation and COX‐2 protein expression were measured respectively after treated with selective COX‐2 inhibitor NS‐398 . Results CCK‐8 assay and plate colony formation assay displayed the proliferation rate of HL 7702/HBx was higher than that of HL7702/Mock and HL7702 cells . The level of COX‐2 mRNA and protein expression was greater in HL7702/HBx cells compared with that of HL7702/Mock and HL7702 cells(P<0 .05) . COX‐2 selective inhibitor NS‐398 suppressed growth of three groups of cells in a time‐dependent manner . The proliferation inhibition rate of HL7702/HBx cells was significantly higher than that of other two groups in three time points respectively (P<0 .05) . After treated with 50 μmol/L NS‐398 for 72 h ,COX‐2 expres‐sion was suppressed in all group , however , this phenomenon is more obvious in HL7702/HBx cells (P<0 .05) . Conclusion Regulate the expression of COX‐2 is one of the pathway for the effect of HBx on the HL7702 proliferation ,COX‐2 inhibitor can suppress the cell proliferation for the greater part in‐duced by this pathway .