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

 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。

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

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 analysis. The change of Myc and MMP-1 was tested by immunohistochemical staining and the expression of LN was tested by radioimmunoassay. Results The CD82 protein expression and apoptosis were up-regulated in the cells transfected by KAI1, the difference is significant (P< 0.05). The expressions of Myc, MMP-1 and LN were descended. Conclusion The suppressor gene KAI1 could inhibit the proliferation of the small cell lung cell and descende the expression of Myc, MMP-1 and LN.