nm23-H1基因转染对膀胱癌T-24细胞转移能力及化疗敏感性的影响

The Effects of nm23-H1 Gene Cell Transfection on Abilities of Invasion,Adhension and Chemotherapeutic Sensitivity of Bladder Cancer T-24 Cell Line

目的探讨nm23-H1基因对人膀胱癌T-24细胞侵袭转移能力及化疗敏感性的影响。方法将nm23-H1真核表达质粒通过电击穿孔转染技术导入人膀胱癌细胞株T-24,G418筛选阳性克隆,免疫组织化学方法检测nm23-H1基因的表达情况。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法检测T-24细胞转染前后对化疗药物敏感性的变化。结果以电穿孔技术将nm23-H1基因转入T-24细胞后,获得稳定表达;免疫组织化学结果显示T-24细胞株在nm23-H1基因转染前后其表达水平有明显差异;转染后T-24细胞侵袭、粘附、趋化运动能力有明显的降低。转染后T-24细胞对顺铂(CDDP)明显增敏。结论nm23-H1基因对人膀胱癌T-24细胞的侵袭转移能力具有显著抑制作用。nm23-H1基因能增强T24细胞对顺铂的化疗敏感性。

Objective To investigate the transfection and expression of nm23-H1 gene in human bladder cancer cell line T-24,and to find out whether nm23-H1 could affect the metastases ability and the chemotherapeutic sensitivity of T-24 cell line.Methods By electroporation way,the pCMV-Bam-neo-nm23-H1 plasmid was transferred into bladder cancer line T-24,and the nm23-H1 vector-producer cell line T-24/nm23-H1 was isolated with G418 selection.The expression of nm23-H1 gene was detected by immunohistochemistry.The T-24 invasion,adherence and migration abilities between transfected and non-transfected cell lines were detected by transwell-room and wash techniques.The difference or change to the sensitivity of chemotherapeutic drug before and after cell transfected was evaluated by MTT.Results The stable expression of nm23-H1 was obtained by pCMV-Bam-Neo system with G418.The expression of nucleoside diphosphate kinase-A（NDPKA） was observed for significant difference between transfected and non-transfected T-24 cell lines;The invasive,adherence and migration abilities of transfected T-24 cells were decreased more than those of non-transfected T-24 （P〈0.05）;The chemo-sensitivity of transfected T-24 line cells to cis-diamminedichloroplatinum（CDDP） increased significantly.Conclusion nm23-H1 gene can inhibit the metastasis of T-24 cell line and increase the chemo-sensitivity of T-24 cell line to CDDP significantly.