KAI1基因抑制胰腺癌细胞鞘氨醇激酶活性及其迁移  

Adenovirus-mediated overexpression of KAI 1 suppresses sphingosine kinase activation and metastasis of pancreatic carcinoma cells

作　　者：郭晓钟[1] 张巍巍[1] 王立生[2] 鲁茁壮[2] 王华[2] 徐建华[1] 田宏[1] 

机构地区：[1]沈阳军区总医院消化科,110016 [2]军事医学科学院放射与辐射医学研究所

出　　处：《中华内科杂志》2006年第9期752-754,共3页Chinese Journal of Internal Medicine

基　　金：国家自然科学基金(30470798)

摘　　要：目的研究复制缺陷型腺病毒介导人肿瘤转移抑制基因 KAI1在胰腺癌细胞中高表达对细胞鞘氨醇激酶(SPK)信号的影响。方法构建重组腺病毒 Ad-KAI1;通过流式细胞技术检测感染人胰腺癌细胞 KAI1的表达;应用[γ-^(32)P]ATP 掺入法和扩散盒检测 SPK 酶活性及胰腺癌细胞的迁移。结果成功构建的 Ad-KAI1可以高效感染人胰腺癌细胞 PANC Ⅰ达86.3%,同时检测到PANC Ⅰ细胞 SPK 高表达,而 Ad-KAI1可以降低 PANC Ⅰ细胞的 SPK 活性,并抑制其迁移。结论高表达的 KAI1基因具有抑制胰腺癌细胞 SPK 活性及其促迁移作用。Objective To investigate influence of KAI1 expression on the pancreatic carcinoma cell migration and its mechanisms. Methods We construct an adenoviral vector carrying the full length cDNA of KAI1 （Ad-KAI1）. The Ad-KAI1 virus was packed and amplified in 293 cells and was purified by AdenoX^TM Virus Purification Kit. The adnovirs-mediated KAI1 expression in human pancreatic carcinoma cell line （PANC Ⅰ ） was detected by flow cytometric analysis. Sphingosine kinase （SPK） activation was determined by γ^-32P] incorporation. And cell migration was assayed by using transwell. Results We constructed Ad- KAI1 vector successfully. The 86. 3% of pancreatic cancer cells PANC Ⅰwere identified and infected by Ad-KAI1 constructed by us. At same time, we found out higher concentration of SPK in the PANC Ⅰ cancer cells without infection of Ad-KAI1 using RT-PCR. Overexpression of KAI1 significantly inhibited the activity of SPK and the migration ability of the PANC Ⅰcells, which was induced by activity of SPK. Conclusions Adnovirus-mediated overexpression of KAI1 suppresses sphingosine kinase activation and migration of pancreatic carcinoma cells. This research is helpful for elucidating the anti-metastasis mechanisms KAI1 in pancreatic carcinoma cells.

关 键 词：胰腺肿瘤 KAI1基因 鞘氨醇激酶 

分 类 号：R735.9[医药卫生—肿瘤]