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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
Objective To investigate the effect of nm23_H1 on the invasion and metastasis of Tca81 1 3 cell line through the transformation of nm23_H1 and the introduction of Tca81 1 3 cell line. Methods Gene transfection technique was used to prepare high purity nm2 3_H1 eukaryotic expression plasmid. The cationic liposome-mediated transfection technique was used to complete the transfection method. The expression of nucleoside diphosphokinase A (NDPKA), a protein of nm 2 3_H1 before and after transfection, was detected by immunohistochemistry. Transwell chamber and scouring experiments were used to observe the changes of invasion, adhesion and chemotaxis motility of cells before and after transfection. The effect of nm2 3_H1 on chemosensitivity of Tca81 1 3 was observed by MTT assay. Results Recombinant pCMV_Neo_Bam eukaryotic expression vector was used to transfect nm2 3_H1 into oral cancer cells and obtain stable expression. Tca81 1 3 cell line before and after transfection nm2 3_H1 gene expression levels were significantly different, Tca81 1 3 transfected cells invasion, adhesion, chemotactic ability were significantly reduced, before and after transfection of Tca81 1 3 cells adriamycin ( ADM), 5-fluorouracil (5_FU) and methotrexate (MTX). The Tca81 1 3 cell line significantly enhanced the sensitivity to cisplatin (CDDP) after transfection. Conclusion nm2 3_H1 has a significant inhibitory effect on the invasion and metastasis of Tca81 1 3 cells. Nm2 3_H1 may enhance the sensitivity of CDDP chemotherapy by specifically affecting the intracellular energy exchange process