实时监测癌细胞死亡早期预测化疗疗效的研究

目的:探讨实时监测癌细胞死亡早期血清中CK18含量预测化疗疗效的可行性。方法:癌细胞死亡后释放的可溶性细胞内大分子可于血清中检出；CK18是上皮细胞的细胞骨架蛋白，CK18-Asp396是CK18在癌细胞凋亡过程中的产物，可被M30抗体特异性检出，M65抗体可检出CK18在癌细胞死亡过程中产生的所有可溶性片段。CK18-Asp396和可溶性CK18总量的变化可用于监测癌细胞的凋亡和死亡。本实验在30例乳腺癌患者中检测化疗前和第1 周期化疗给药结束后24～48h 血清中CK18-Asp396和可溶性CK18总量；采用卡方检验分析第1 周期化疗前后CK18-Asp396和可溶性CK18总量变化率与4 周期后化疗疗效之间的关系。结果:30例乳腺癌患者中，ⅡB 期2 例，ⅢA 期11例，ⅢB 期1 例，ⅢC 期2 例，Ⅳ期14例，均接受以蒽环类和/或紫杉类为基础的联合化疗，4 周期后按RECIST标准进行疗效评价，共有3 例完全缓解，19例部分缓解，7 例稳定，1 例进展，有效率为73.3%（22/30）。 化疗前和第1 周期给药结束24～48h 血清CK18-Asp396变化率>20% 和/或CK18总量变化率>30% 的19例患者均为缓解，而其余的11例患者中仅3 例部分缓解，7 例稳定，1 例进展（χ2=18.843，P=0.001）。 结论:乳腺癌患者化疗前和第1 周期给药结束24～48h 血清中不同片段CK18含量的变化率与化疗疗效明显相关，作为预测乳腺癌及其它上皮来源恶性肿瘤化疗疗效的生物学指标具有进一步研究的价值。 

Monitoring Tumor Cell Death in Real-time Systems for Early Determination of Response to Chemotherapy in Breast Cancer

Objective:To explore the possibility of monitoring tumor cell death in real-time for early determination of re -sponse to chemotherapy. Methods:Intracellular macromolecules are released from dying tumor cells and can subsequently be detected in patient blood. The M30antibody was used to detect the caspase-degraded product of the cytoskeletal pro -tein CK 18denoted as CK 18-Asp396 during apoptosis of epithelial cells, and the M65antibody was used to detect all of the products of CK 18during apoptosis and necrosis. The change in CK18-Asp396 and total soluble CK18levels could be used to monitor the apoptosis and death of tumor cells. Sera from 30patients with breast cancer were collected at24-48hours before and after the 1st cycle of chemotherapy and the content of CK18-Asp396 and total soluble CK18was quantitatively examined with ELISA assays. Correlations between the change in CK18and efficacy were determined by χ2 test. Results: Of the 30patients, there were 2 patients with stageⅡB, 11with stage ⅢA, 1 with stage ⅢB, 2 with stage ⅢC and 14with stage Ⅳdisease. All30patients received a combined chemotherapy regimen based on anthracyclines and/or taxans. After 4 cycles, RECIST criteria were used to evaluate the response. Three patients had a complete response, 19patients had partial response, 7 had stable disease and 1 had progressive disease. The response rate was 73.3 % (22/30). All of the 19 patients who displayed changes in caspase-cleaved CK 18>20% and/or change in total soluble CK18>30% responded. Among the other 11patients, 3 patients had PR, 7 had SD and 1 had PD (χ2= 18.843 , P = 0.001 ). Conclusion:There is a significant relationship of increase in caspase-cleaved CK 18and total soluble CK18 level in serum with overall response to therapy. CK18-Asp396 and total soluble CK18might be used as a novel biomarker for early prediction of response to che-motherapy in patients with breast cancer or other malignant tumors.