nm23-H1基因对Tca8113的转移能力及化疗敏感性影响的实验研究

The Effects of nm23-H1 on Metastases Ability and Chemo-sensitivity in Tca8113 Cell Lines

目的 通过nm2 3_H1的转化及导入Tca81 1 3细胞株 ,建立稳定、高效、低毒的转染方法 ,观察nm2 3_H1对Tca81 1 3细胞株侵袭转移能力的影响。方法 利用基因转化技术 ,制备高纯度的nm2 3_H1真核表达质粒。利用阳离子脂质体介导的转染技术 ,完成转染方法的建立。利用免疫组化技术 ,检测转染前后的nm2 3_H1的蛋白产物核苷二磷酸激酶A (NDPKA)的表达。利用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。MTT法观察nm2 3_H1对Tca81 1 3化疗敏感性影响。结果 使用重组的pCMV_Neo_Bam真核表达载体 ,将nm2 3_H1转染口腔癌细胞 ,并获得稳定表达。Tca81 1 3细胞株nm2 3_H1基因转染前后表达水平有明显差异 ,转染后Tca81 1 3细胞侵袭、粘附、趋化运动能力均显著降低 ,转染前后Tca81 1 3细胞株对阿霉素 (ADM)、5_氟尿嘧啶 (5_FU)、甲氨喋呤 (MTX)的化疗敏感性无显著差异 ,转染后Tca81 1 3细胞株对顺铂 (CDDP)明显增敏。结论 nm2 3_H1对Tca81 1 3细胞的侵袭转移能力具有显著抑制作用 ,nm2

Objective\ The purpose of this study was to establish a stable, high_efficient and low_toxic way of transfecting nm23_H1 into Tca8113 line cells, and then to find out whether nm23_H1 could affect the invasion and metastases ability of Tca8113 line cells.Methods\ nm23_H1 was transfected into Tca8113 line cells with Lipofect. The different expressions of nm23_H1 between transfected and non_transfected line cells were detected by the method of immunohistochemistry. The difference of the invasion and metastases ability between transfected and non_transfected line cells was detected by transwell_room and wash techniques. The change of chemo_sensitivity was evaluated by MTT.Results\ Using pCMV_NEO_BAM system to keep stable expression of nm23_H1, the significant difference of NDPKA expression between transfected and non_transfected Tca8113 line cells was discovered; The metastases ability of transfected Tca8113 line cells decreased significantly;The chemo_sensitivity of transfected Tca8113 line cells to CDDP increased significantly.Conclusion\ nm23_H1 can inhibit the metastases of Tca8113 line cells and increase the chemo_sensitivity to CDDP significantly.\;