nm23-h1转染腺样囊性癌细胞及其增殖能力的实验研究

目的　考察nm23-h1导入腺样囊性癌细胞(ACC-M)后对其体内、外增殖的影响。方法　以阳离子脂质体 (lipofectamine)介导nm23-h1转染ACC-M细胞,体外培养并用G418培养基筛选,体外观察转染组生长情况,并用细胞爬片免疫组化ABC法染色Ki67单克隆抗体;将nm23-h1阳性表达的ACC-M植入10只裸鼠皮下,同时以未转染的 ACC-M作对照,观察肿瘤大小。4周后处死动物,获取标本,常规固定、包埋、切片后采用免疫组化LsAB法研究Ki67 的表达并用流式细胞仪对转染组和非转染组的移植瘤进行细胞周期时相分析。结果　体外培养转染组细胞生长较未转染组慢,这种趋势随着转染组传代次数的增加而更加明显,Ki67呈阳性表达,未转染组Ki67呈部分阳性表达;转染组ACC-M体内移植瘤早期(2周)生长缓慢,2周后生长无统计学差异(P>0.05),细胞周期时相分析结果与之一致。组织切片显示瘤体Ki67,nm23-h1表达均为阴性。结论　nm23-h1的导入可能对ACC-M细胞增殖具有短时间的抑制作用。

In vitro and in vivo Study on Proliferation of Adnoid Cystic Carcinoma Cell Lines after nm23-h1 Introduction

Objective To study the effect of nm23-h1 transfection on proliferation characteristics of adnoid carcinoma cell lines in vitro and in vivo.Methods In vitro ACC-M cell lines were incubated after putative anti-metastatic gene nm23-h1 was introduced into the cells with the help of G418 selective incubation base. The ACC-M cells were transplanted into 10 BALB/C nude mices subcutaneously and non-transfected cell lines were taken as control. Immunohisto chemistry and Ki67 antibody were employed to study the proliferation character of cell crawling pieces and paraffin-bedded slice, meanwhile, the solid tumor of both groups were prepared for flow cytometry(FCM).Results Transfected cells grew slower than non-transfected cells and this trend became more obvious as passages passed on. In vitro the expression of Ki67 of transfected cells was little stronger than non-transfected cells, while the expression of Ki67 in solid slices was almost negative in both groups. Transfected cells presented slower growth than non-transfected cells in the early stage(2 weeks) and 2 weeks later there was no obvious difference in size(P>0.05). FCM value accorded well with the result.Conclusion Introduction of nm23-h1 into the ACC-M cell lines may have transient inhibitory effects on its proliferation.