Oncoprotein 18在肝癌侵袭转移过程中的作用  

Role of Oncoprotein 18 in metastasis of hepatocellular carcinoma

作　　者：甘淋[1] 段承刚[1] 龚舒[1] 郭坤[2] 舒宏[2] 刘银坤[2] 

机构地区：[1]泸州医学院医学基础研究中心,四川省泸州市646000 [2]复旦大学中山医院肝癌研究所,上海市200032

出　　处：《世界华人消化杂志》2012年第23期2151-2156,共6页World Chinese Journal of Digestology

基　　金：国家重大专项基金资助项目;No.2008ZX10002-021~~

摘　　要：目的:探讨原癌蛋白18(Oncoprotein 18,Op18)在肝癌侵袭转移过程中的作用机制.方法:采用R N A干扰,抑制人肝癌细胞HCCLM3中Op18的表达,RT-PCR和Westernb l o t评价干扰效率;通过细胞粘附分析、体外Transwell分析法检测Op18表达缺失后对HCCLM3细胞粘附、运动和侵袭能力的改变;RT-PCR和免疫组织化学分别在48例未伴转移的肝癌组织和48例伴转移的肝癌组织中解析Op18表达与肝癌侵袭转移的关系.结果:RT-PCR和Western blot结果显示:在HCCLM3细胞中行RNAi后Op18表达被有效抑制,抑制率达到80%以上;Op18表达缺失后,在不同时间点(20、40和60 min)实验组细胞粘附能力(0.616±0.057、0.740±0.0713和1.001±0.083)较阴性对照组(0.944±0.068、1.196±0.115和1.441±0.053)明显下降(P<0.05);Transwell实验结果提示:经RNAi后实验组细胞运动、侵袭能力(运动:0.145±0.011,侵袭0.127±0.008)较阴性对照组(运动:0.206±0.008,侵袭:0.168±0.012)显著降低(P<0.01);分别在伴转移和未伴转移的肝癌组织中检测Op18的表达,RT-PCR(Op18与GAPDH相对比值:0.560±0.128vs 0.414±0.086)和IHC(积分吸光度值为:624.771±100.032 vs 413.786±71.833)实验结果均提示Op18在伴转移的肝癌组织中的表达更高(P<0.01).结论:Op18在肝癌的侵袭转移过程中发挥重要作用.AIM： To explore the role of Oncoprotein 18（Op18） in metastasis of hepatocellular carcinoma（HCC）.METHODS： Op18 expression was repressed in HCCLM3 cells by RNA interference,and interference effect was evaluated by RT-PCR and Western blot.Cell adhesion,migration and invasion were analyzed by cell adhesion assay and Transwell assay in vitro.Op18 expression was detected by RT-PCR and immunohistochemistry in 96 HCC specimens with or without metastasis.RESULTS： Op18 expression was effectively inhibited by RNA interference in HCCLM3 cells.Cell adhesion was significantly suppressed inthe RNAi group compared to the mock group（20 min： 0.616 ± 0.057 vs 0.944 ± 0.068;40 min： 0.740 ± 0.0713 vs 1.196 ± 0.115;60 min： 1.001 ± 0.083 vs 1.441 ± 0.053;all P 〈 0.05）.Transwell assay revealed that cell migration and invasion decreased in the RNAi group compared to the mock group（migration： 0.145 ± 0.011 vs 0.206 ± 0.008;invasion： 0.127 ± 0.008 vs 0.168 ± 0.012;both P 〈 0.01）.Op18 expression was detected in 96 HCC tissues with or without metastasis.RTPCR demonstrated that Op18 was overexpressed in HCC tissues with metastasis（n = 48） compared with HCC tissues without metastasis（n = 48,Op18/GAPDH relative ratio： 0.560 ± 0.128 vs 0.414 ± 0.086）;and IHC results also indicated that Op18 expression was up-regulated in HCC tissues with metastasis in comparison with HCC tissues without metastasis（integrated density： 624.771 ± 100.032 vs 413.786 ± 71.833,P 〈 0.01）.CONCLUSION： Overexpression of Op18 may play an important role in HCC metastasis.

关 键 词：肝细胞癌 原癌蛋白18 RNA干扰 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]