脂质体介导KAI1基因转染对小细胞肺癌NCI-H446细胞株的抑制作用

Study of the Inhibition of Small Cell Lung Cancer with Transfection the Gene KAI1

摘要: 目的　探讨新抑癌基因KAI1对小细胞肺癌NCI H4 4 6细胞株抑制增殖和浸润转移的作用。方法　用脂质体介导的基因转染方法, 借助真核质粒表达载体 (PCMV NEO XhoI), 将抑癌基因KAI1转入小细胞肺癌NCI H4 4 6细胞中, 经G4 18筛选, 获得稳定表达的细胞克隆。观察NCI H4 4 6细胞的生长, MTT法检测细胞体外增殖能力, 流式细胞仪进行细胞凋亡分析, 间接免疫荧光染色结合流式细胞仪检测转染前后细胞CD82蛋白的表达。免疫组化测定Myc及MMP 1(基质金属蛋白酶 1)的表达, 放射免疫测定LN(层黏蛋白 )表达。结果　脂质体 KAI1基因转染的小细胞肺癌细胞CD82表达增加, 细胞增殖能力下降, 凋亡增加, 癌基因Myc, MMP 1及LN表达下降。结论　抑癌基因KAI1抑制小细胞肺癌NCI H4 4 6细胞株的增殖, 降低Myc, MMP 1及LN的表达。

Abstract: Objective To study the inhibition of proliferation and metastasis of the small cell lung cell with transfection of a new-defined tumor metastasis suppressor gene KAI1. Methods To establish NCI-H446 cell line with stably expressing tumor metastasis suppressor gene KAI1,the gene KAI1 was transfected into NCI-H446 cell by the vector(PCMV-NEO-XhoI).We got clone cells after selected by G418. MTT methods examine the cell proliferation. The CD82 protein expression and apoptosis was detected by cytometry analysi...