顺铂和放疗联合应用在人喉鳞癌细胞中相互作用的研究

Mutual Effect of Cisplatin and Radiation Used in Combination in Hep-2 Cells

目的 研究CDDP和放疗联合应用于人喉鳞状细胞癌细胞的相互作用,以探讨放、化疗两种处理方式应用的顺序及与端粒酶活性的关系。方法 Hep 2 细胞分别经放化疗不同剂量及顺序处理,实验设对照组、CDDP组、放疗组、CDDP+放疗组及放疗+CDDP 组,分别用 MTT 法、端粒酶活性检测PCR ELISA法及聚丙烯酰胺凝胶电泳及银染法分析各种处理后的细胞存活率及端粒酶活性的变化。结果 CDDP+放疗组比同等处理剂量的放疗+CDDP组的细胞存活率低,且有显著差异。CDDP及放疗均能使端粒酶活性明显增高,并表明为浓度依赖性。但在加入另一种处理后,端粒酶活性降低,其降低的程度与细胞存活率的变化一致(r=0.9445,b=13.61)。结论 对Hep-2 细胞先化疗后放疗的治疗效果优于先放疗后化疗,联合治疗后的端粒酶活性可反映治疗效果。

Objective To study the relationship between Cisplatin (CDDP) and radiation used in combination in human laryngeal squamous cells (Hep-2) , in order to determine the optimum sequence of the two treatments, and the relations with telomerase activity.Methods Hep-2 cells were exposed to CDDP and ^(60)Co gamma-irradiation with different dose and sepuence. Our group cells as follows : untreated group、CDDP group、irradiation group、CDDP + irradiation group and irradiation + CDDP group. MTT assay、PCR based Telomeric Repeat Amplification Protocol (TRAP) coupled with ELISA and polyacrylamide gel electrophoresis (PAGE) with sliver staining were used to estimate the cell survival rate and the telomerase activity (TA) before and after treatments. Results The cell survival rate of the group treated by CDDP plus radiation was lower than that by irradiation plus CDDP with significant statistical difference. Analysis of telomerase activity indicated that both CDDP and irradiation could independently increase telomerase activity in a dose dependent manner, however, addition of the other treatment decreased TA, decreased range scale was consistent with change of cell survival rate.Conclusion The treatment of Hep-2 cell by chemotherapy plus irradiation is superior to that treatment by irradiation plus chemotherapy: after combined treatments, TA can reflect the effects of therapies.