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
作者：陈绍维,黄洪章,潘剑,温玉明,廖湘凌,王昌美,李芃,杨凯
Author：CHEN Shaowei*, HUANG Hongzhang, PAN Jian,et al.
收稿日期：2003-06-25 年卷（期）页码：2003,21(03):180-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：nm23-H1,Tca8113,转移,化疗敏感性,
Key words：nm23-H1,Tca8113 cell lines,metastasis,chemo-sensitivity,
基金项目：
国家自然科学基金 (编号 39870746);福建省自然基金重点项目 (编号C0220002)资助项目
中文摘要
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 3-H1可能通过特异性地影响细胞内的能量交换过程来达到对CDDP化疗增敏的效果。
英文摘要
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.
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