间隙连接蛋白Connexin32对肝癌侵袭和转移能力的影响  

Aberrantly localized connexin32 protein enhances invasion and metastasis of hepatocellular carcinoma cells in vivo

作　　者：李庆昌[1] 谢成耀[1] 高辉[1] 徐洪涛[1] 邱雪杉[1] 王恩华[1] 

机构地区：[1]中国医科大学基础医学院病理教研室中国医科大学附属第一医院病理科,辽宁沈阳110001

出　　处：《中华肿瘤防治杂志》2009年第8期561-564,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30700806)

摘　　要：目的:探讨肝癌细胞中间隙连接蛋白Connexin32对肝癌细胞体内侵袭和转移能力的影响。方法:利用可以通过增减细胞外环境中Doxycycline调控Connexin32蛋白表达的肝癌细胞HuH7Tet-off Cx32亚克隆,将之原位移植到重度联合免疫缺陷鼠肝脏浆膜下,通过饮用水中添加(去除)Doxy-cycline调控动物体内Connexin32的表达,8周后处死小鼠,观察肿瘤形成及转移情况,蛋白质印迹法及免疫组织化学方法检测其与对照组的Connexin32蛋白表达情况。结果:移植HuH7Tet-off Cx32细胞的小鼠,在饮用水中去除Doxycycline诱发Connexin32高表达条件下,形成明显的肝内转移灶和腹膜转移灶;而饮用水中添加了Doxycycline的小鼠没有发现明显的转移灶形成。免疫组化证实,高表达的Connexin32蛋白定位于细胞质。蛋白质印迹法结果显示,Doxycycline在小鼠体内呈现较好的诱导性。结论:细胞质内异位蓄积的Connexin32蛋白明显增强肝癌细胞的体内侵袭和转移能力。OBJECTIVE： To investigate the effects of gap junction protein eonnexin32 on invasiveness and metastasis of hepatocellular carcinoma cells in vivo. METHODS：The previously established HuH7 Tet off Cx32 subclone, in which connexin32 expression could be controlled by supplement/removal of doxyeycline, was in situ transplanted to the liver subserosa of severe combined immunodeficiency mice. The expression level of connexin32 was controlled by doxycycline supplemented in drinking water. Eight weeks later, the mice were sacrificed and the status of tumorgeneeity and metastasis was carefully examined. The connexin32 expression was detected with both Western-blot and immunohistochemical staining. RESULTS： The mice injected with HuH7 Tet off Cx32 cells under circumstance of doxycycline depletion and connexin32 overexpression were found to form obvious intrahepatic and peritoneal metastses, while no metastases was found in mice supplemented with doxycycline thereafter the connexin32 expression was inhibited. The immunohistochemical study showed connexin32 was expressed in cytoplasm and the Western-blot results demonstrated that high inducibility of connexin32 expression was maintained in the xenografts. CONCLUSION：Cytoplasmic accumulation of eonnexin32 protein can promote tumor invasion and metastasis in hepatocellular carcinoma in vivo.

关 键 词：肝肿瘤 间隙连接 肿瘤转移 肿瘤侵润 

分 类 号：R735.7[医药卫生—肿瘤]