Latexin基因转染对CD133^＋MIAPaca-2胰腺癌干细胞增殖的影响  

Effect of Latexin gene transfection on proliferation in CD133＋ MIAPaca-2 pancreatic cells

作　　者：王成[1] 蔡振寨[1] 周羽翙 郑继行[1] 薛战雄[1] 

机构地区：[1]温州医科大学附属第二医院消化科,浙江温州325000

出　　处：《中华胰腺病杂志》2016年第1期10-14,共5页Chinese Journal of Pancreatology

基　　金：浙江省自然科学基金（Y2100546）

摘　　要：目的探讨Latexin（Lxn）基因对CD133＋MIAPaca-2胰腺癌干细胞增殖的影响。方法采用磁珠分选法从MIAPaca-2胰腺癌细胞株中分选出CD133＋细胞，在无血清培养基中观察其形态变化及种植到裸鼠皮下后的成瘤能力。采用脂质体法将不同浓度的插入Lxn基因（1、3、5μg）的质粒转染CD133＋MIAPaca-2胰腺癌细胞。应用RT—PCR及蛋白质印迹法检测转染的癌细胞LxnmRNA及蛋白表达量。CCK-8法检测转染癌细胞的增殖能力。结果成功分离出CD133＋；MIAPaca-2胰腺癌细胞，它以悬浮成球的方式生长，以1×10^5个癌细胞种植裸鼠皮下的成瘤率达100％。转染插入Lxn的质粒后，CD133＋ MIAPaca-2胰腺癌细胞Lxn基因的表达呈转染剂量依赖性地增加，转染5μg质粒的细胞LxnmRNA及蛋白表达量分别为20．80±0．98、16．80±2．73，显著高于未转染细胞的1．02±0．01、1．01±0．01，差异均有统计学意义（P值均〈0．05）；转染后癌细胞增殖活性呈转染剂量及培养时间依赖性地被抑制，转染0．4μg质粒的癌细胞的生长抑制率达36．2％，显著高于未转染细胞的抑制率，差异有统计学意义（P〈0．05）。结论CD133＋MIAPaca-2胰腺癌细胞具有肿瘤干细胞部分特性，Lxn基因转染对CD133＋MIAPaca-2胰腺癌干细胞增殖具有抑制作用。Objective To explore the effect of Latexin （Lxn） gene transfection on proliferation of CD133＋ MIAPaca-2 pancreatic cancer stem-like cells. Methods CD133＋ MIAPaca-2 cells were isolated and sorted by magnetic activated cell sorting from pancreatic cancer MIAPaca-2 cell line. CD133＋ MIAPaca-2 cells were cultured in serum-free medium and the capacity for proliferation, and tumorigenicity of CD133 MIAPaca-2 cells was determined by the floating spheres test and tumor xenograft assays. The CD13＋ MIAPaca-2 cells were transfected with Lxn plasmid （ 1, 3, 5 μg）. After transfection, the protein and mRNA expression of Lxn in CD133＋and CD133-MIAPaca-2 cells were detected by Western blotting and RT-PCR, respectively. Cell proliferation was assayed by CCK-8. Results CD133 MIAPaca-2 cells were successfully isolated, and it grew into a ball-suspended way, the tumorigenicity rate in nude mice with subcutaneous injection 1 × 105 cancer cells was 100%. After Lxn plasmid transfection, the expression of Lxn in CD133＋ MIAPaca-2 cells was increased in a dose dependent manner, the Lxn protein and mRNA expression of tumor cells transfected with 5 μg plasmid was 20.80 ±0.98, 16.80 ±2.73, which was significantly higher than that in non-transfected cells （1.02 ±0.01, 1.01 ± 0.01 ） , and the difference between the two groups was statistically significant （ P 〈 0.05 ）. After transfeetion, cellular proliferation activity also showed a transfection dose and culture time-dependent decrease,the inhibition rate of tumor cells transfected with 0.4 μg plasmid was 36.2%, which was significantly different from that in non-transfected cells （P 〈 0.05 ）. Conclusions CD133＋ MIAPaca-2 pancreatic cancer cells have some characteristics of cancer stem cells. Lxn gene transfection can inhibit the proliferation of CD133＋ MIAPaca-2 cells.

关 键 词：胰腺肿瘤 肿瘤干细胞 Lxn基因 抗原 CD133 细胞增殖 

分 类 号：R735.9[医药卫生—肿瘤]