EP方案化疗对广泛期小细胞肺癌CTCs上皮-间质转化影响的初步研究

Preliminary Study on Effect of EP Regimen on Epithelial-mesenchymal Transition of CTCs in Patients with Extensive Small Cell Lung Cancer

目的:研究EP方案化疗对广泛期小细胞肺癌(ED-SCLC)患者外周血循环肿瘤细胞(CTCs上皮-间质转化(EMT)的影响。方法:将2016年10月-2017年12月赣州市人民医院收治的76例符合入组条件的ED-SCLC受试者作为本研究对象,采集分离获得ED-SCLC受试者外周静脉血中CTCs,运用密度梯度离心法和流式细胞术富集分析化疗前后CTCs数目及细胞表型情况;采用实时荧光定量PCR技术(qRT-PCR)分析化疗前后细胞角蛋白-19(CK-19)、钙离子依赖型细胞黏附素家族(E-cadherin)、波形蛋白(Vimentin)的m RNA相对阳性表达情况及其变化情况。采用酶联免疫吸附测定法(ELISA)测定CK-19、E-cadherin及Vimentin蛋白水平变化情况。结果:用EP方案化疗后的ED-SCLC患者的CTCs计数与阳性表达例数较化疗前均明显降低(P<0.05);CK-19、E-cadherin的mRNA相对阳性表达率较化疗前均明显降低,而Vimentin的m RNA相对阳性表达率较化疗前升高,差异均有统计学意义(P<0.05)。患者E-cadherin、Vimentin蛋白水平较化疗前均明显降低,而CK-19蛋白水平较化疗前升高(P<0.05)。结论:ED-SCLC患者采用EP方案化疗后可能通过抑制CTCs发生EMT过程,进而产生抑制肿瘤复发及转移效果,这对进一步明确SCLC复发、转移之间的量化对应关系及患者个体化治疗均有积极意义。

Objective: To study the effect of EP regimen on epithelial-mesenchymal transition(EMT) in peripheral blood circulating tumor cells(CTCs) in patients with extensive stage small cell lung cancer(ED-SCLC). Method: Seventy-six ED-SCLC subjects who met the enrollment criteria admitted to the People’s Hospital of Ganzhou City from October 2016 to December 2017 were included in the study. The venous venous blood was collected from ED-SCLC subjects. CTCs were enriched by density gradient centrifugation and flow cytometry to analyze the number of CTCs and cell phenotypes in patients before and after chemotherapy. Real-time quantitative PCR(qRT-PCR) was used to analyze cytokeratin-19(CK-19) before and after chemotherapy, the relative positive expression of mRNA of calcium-dependent cell adhesin family(E-cadherin) and Vimentin and their changes. The levels of CK-19, E-cadherin and Vimentin protein were measured by enzyme-linked immunosorbent assay(ELISA). Result: The number of CTCs and positive expression in ED-SCLC patients treated with EP regimen were significantly lower than those before chemotherapy(P<0.05). The relative positive expression rates of CK-19 and E-cadherin mRNA were significantly lower than those of before chemotherapy, the relative positive expression rate of Vimentin mRNA was higher than that of before chemotherapy, and the differences were statistically significant(P<0.05). The protein levels of E-cadherin and Vimentin were significantly lower than those of before chemotherapy, while the level of CK-19 protein was higher than that of before chemotherapy(P<0.05). Conclusion: ED-SCLC patients may have EMT by inhibiting CTCs in patients after chemotherapy with EP regimen, and then has the effect of inhibiting tumor recurrence and metastasis. The process has positive implications for further clarifying the quantitative correspondence between SCLC recurrence and metastasis and individualized treatment of patients.