nm23-H1基因对口腔癌Acc-M细胞株的转移能力及化疗敏感性影响的体外研究

in vitro Study of the Effect of nm23-H1 on Metastasis Ability and Chemo-sensitivity of Acc-M Cell Lines

目的 通过 nm2 3- H1的转化及导入 Acc- M细胞株 ,建立稳定、高效、低毒的转染方法 ,观察 nm2 3- H1对 Acc- M细胞株侵袭转移能力及化疗敏感性的影响。方法 采用基因转化技术 ,制备高纯度的 nm2 3- H1真核表达质粒。用阳离子脂质体介导的转染技术 ,将 nm2 3- H1基因转染到口腔腺样囊性癌 Acc- M细胞株。用免疫组化技术检测转染前后的 nm2 3- H1的表达。并用 transwell小室和冲刷实验 ,观察转染前后细胞的侵袭、粘附、趋化运动能力的变化。采用 MTT法观察 nm 2 3- H1对 Acc- M化疗敏感性影响。结果 1使用重组的 p CMV - Neo- Bam的真核表达载体 ,将 nm 2 3- H 1转染口腔癌细胞 ,并获得稳定表达。 2 Acc- M细胞株 nm2 3- H 1基因的转染前后表达水平有明显差异。 3转染后的 Acc- M细胞侵袭、粘附、趋化运动能力均显著降低。 4转染前后使用 C- DDP的 L D50 分别为2 .6 4± 0 .4 7,1.85± 0 .4 1(P<0 .0 1)。结论 nm2 3- H1对 Acc- M细胞的侵袭转移能力具有显著抑制作用 。

Objective To transfect nm23 H1 into the Acc M cell lines in a safe, high efficiency and low toxicity way, and then to find out whether nm23 H1 affects the metastasis ability and chemo sensitivity of Acc M cell lines. Methods Lipofect was used to transfect nm23 H1 into Acc M cell lines. The difference in expression of nm23 H1 between the transfected and non transfected cell lines was detected by immunohistochemistry. Then by use of transwell room and wash way, the difference in invasion and metastasis ability between the transfected and non transfected cell lines was tested. MTT method was adopted in finding the change of chemo sensitivity. Results Using pCMV Neo Bam system, we observed the stable expression of nm23 H1 and the significant difference in Nucleoside Diphosphate kinase A, expression between the transfected and non transfected Acc M cell lines. The metastasis ability of transfected Acc M cell lines decreased significantly. The chemo sensitivity of transfected Acc M cell line to cis diamminedichloroplatin (C DDP) increased significantly. Conclusion nm23 H1 can inhibit the metastases of Acc M cell lines significantly and can increase the chemo sensitivity to C DDP significantly.