多种胃癌细胞株中切除修复交叉互补基因1与顺铂作用的关系

Correlation between excision repair cross-complementing gene 1 and sensitivity of gastric cancer celllines to cisplatin

目的探讨6种胃癌细胞株中切除修复交叉互补基因1（ERCCI）与顺铂作用的关系。方法常规培养6种胃癌细胞株；SYBRGreen实时定量聚合酶链反应（Real—timePCR）法及Westernblot法检测6种胃癌细胞株的ERCCImRNA及蛋白的表达；不同浓度顺铂作用6、12、24、48h后，细胞计数试剂盒（CCK一8）法检测细胞存活率，并计算半数抑制浓度（Ic，。），Spearman等级相关分别计算ERCClmRNA、蛋白表达与顺铂作用的相关性。结果6种胃癌细胞株ERCClmRNA的相对表达量为1．40±0．12、1．354-0．12、3．91±0．21、2．92±0．21、4．45±0．14、2．26±0．41，ERCCl蛋白表达灰度分析值为0．58±0．04、0．33±0．07、1．07±0．13、0．40±0．05、1．38±0．17、0．79±0．11，顺铂半数抑制浓度（IC50，mg／L）分另0为36．88±1．29、39．89±1．21、49．72±1．12、44．65±1．05、63．12-4-0．67、54．39±0．98（12h）；4．18±0．14、4．50±0．11、6．23±0．27、5．06±0．22、12．05-4-0．17、5．85-4-0．06（24h）；2．99±0．09、2．86±0．09、4．70±0．13、3．38±0．07、7．86±0．29、3．54±0．03（48h），ERCClmRNA与顺铂IC50的等级相关系数分别为0．89（24h）、0．94（48h）。结论ERCCImRNA的表达与顺铂作用呈等级相关，ERCCl蛋白表达与顺铂作用无相关，ERCClmRNA可应用于临床试验来预测胃癌患者对顺铂的敏感性。

Objective To investigate the correlation between excision repair cross-complementing gene 1 （ ERCC1 ） and sensitivity to cisplatin in 6 kinds of gastric cancer cell lines. Methods Gastric canc- er cell lines were cultured routinely with complete RPMI 1640 medium except AGS cells were cultured with DMEM/F12 medium. The expression of ERCC1 mRNA and protein was detected by using SYBR Green real- time quantitative polymerase chain reaction （Real-time PCR） and Western blotting. All the cell lines were treated with different concentrations of cisplatin for 6, 12, 24 and 48 h, then the ICs0 was calculated respec- tively. For the correlations, the nonparametrie Spearman g p （r） was calculated. Results Relative expres- sion quantity of ERCC1 mRNA in 6 gastric cancer cell lines was 1.40 ± 0. 12, 1.35 ± 0. 12, 3.91± 0. 21, 2. 92 ± 0. 21, 4. 45 ± 0. 14 and 2. 26 ± 0. 41, and the relative expression quantity of ERCC1 protien was 0. 58 ±0. 04, 0. 33 ＋0. 07, 1.07 ±0. 13, 0. 40 ±0. 05, 1.38 ±0. 17 and 0. 79 ±0. 11. ICs0 of DDP （mg/L） was 36. 88 ＋ 1.29, 39.89 ± 1.21, 49.72 ± 1.12, 44.65 ±- 1.05, 63.12 ±0.67 and 54.39 ±0.98 （12 h）, 4. 18 ±0. 14, 4.50 ~0. 11,6. 23 ±0. 27, 5.06 ±0. 22, 12. 05 ±0. 17 and 5.85 ±0. 06 （24 h）, and 2.99 ±0. 09, 2.86 ±0. 09, 4. 70 ±0. 13,3.38 ±0. 07, 7.86 ＋0. 29 and 3.54 ±0. 03 （48 h） respec- tively. Correlation coefficients between ERCC1 mRNA and ICs0 of DDP were 0. 89 （ 24 h ） and 0. 94 （48 h） respectively. Conclusion There was a correlation between ERCC1 mRNA level and sensitivity to cisplatin, but no correlation between protein level and cisplatin. ERCC1 mRNA can be a feasible predicted factor applied in clinical research for patients with gastric cancer treated with cisplatin-based regimens.