不同放化疗敏感性的食管癌细胞株中MGMT和Survivin的表达

Expression of MGMT and Survivin in esophageal squamous cancer cell lines susceptible to chemo-radiosensitivity

目的比较3株食管鳞癌细胞株对放化疗的敏感性,并检测O^6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)和生存蛋白(Survivin)在其中的表达情况。方法 MTT实验检测3株细胞系在不同浓度顺铂和氟尿嘧啶药物中的存活分数,并利用半数抑制浓度(IC50)值评估细胞株对化疗药物的耐受性;克隆形成实验检测3株细胞系在不同剂量照射后的存活分数,并利用单击多靶模型拟合剂量存活曲线计算放射生物学参数D0、Dq、N、SF2以评价3株细胞系对放射线的敏感性;Western blot和qRT-PCR检测3株细胞系中MGMT和Survivin的表达。结果 TE-1在顺铂和氟尿嘧啶中的IC50值高于KYSE-70;TE-1较KYSE-70对放射线耐受(P<0.05)。Western blot和qRT-PCR显示MGMT和Survivin的表达在TE-1中均高于KYSE-70,差异有统计学意义(P<0.05)。结论MGMT和Survivin在对化疗和放疗耐受的细胞株中表达水平较高,在对化疗和放疗敏感的细胞株中表达较低,这为更好地研究食管鳞癌放化疗耐受机制提供了实验依据。

Objective To detect the radiosensitivity and chemosensitivity in different esophageal squamous cancer cells, and compare the MGMT and Survivin protein expression. Methods MTT assay was used to detect the viabili- ty of three cell lines in different concentrations of cisplatin and fluorouracil, and ICs0 value was used to evaluate the tolerance of the cell lines to chemotherapeutic drugs. Clone formation assay was used to valuate the radiosensitivity of three cell lines to the radiation by the radiation biological parameters of DO, Dq, N, SF2 of the multiple target model. Western blot and qRT-PCR was used to detect the expression of three cell lines. Results ICs0 value of TE- l was higher than that of KYSE-70 in cisplatin and 5-fluorouracil. TE-1 was more tolerant of radiation compared with KYSE-70（P 〈0. 05）. Western blot and qRT-PCR showed that the expression of MGMT and Survivin in TE-1 were higher than that of KYSE-70 （P 〈 0.05 ）. The difference was statistically significant. Conclusion The expres- sion of MGMT and survivin in chemotherapy-resistant and radiotherapy-resistant cell lines is higher than that in the chemotherapeutic and radiotherapy-sensitive cell lines, which provide a better way to study the chemoradiation tol- erance mechanism of esophageal squamous cell carcinoma.