中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
2期
245-247
,共3页
周鑫%郑春宁%刘国勤%王希%孙少川
週鑫%鄭春寧%劉國勤%王希%孫少川
주흠%정춘저%류국근%왕희%손소천
胃癌%顺铂%切除修复交叉互补基因1
胃癌%順鉑%切除脩複交扠互補基因1
위암%순박%절제수복교차호보기인1
Gastric cancer%Cisplatin%Excision repair cross-complementing gene 1
目的 探讨6种胃癌细胞株中切除修复交叉互补基因1(ERCC1)与顺铂作用的关系.方法 常规培养6种胃癌细胞株；SYBR Green实时定量聚合酶链反应(Real-time PCR)法及Western blot法检测6种胃癌细胞株的ERCC1 mRNA及蛋白的表达；不同浓度顺铂作用6、12、24、48 h后,细胞计数试剂盒(CCK-8)法检测细胞存活率,并计算半数抑制浓度(IC50),Spearman等级相关分别计算ERCC1 mRNA、蛋白表达与顺铂作用的相关性.结果 6种胃癌细胞株ERCC1 mRNA的相对表达量为1.40±0.12、1.35±0.12、3.91 ±0.21、2.92±0.21、4.45 ±0.14、2.26±0.41,ERCC1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,mag/L)分别为36.88±1.29、39.89±1.21、49.72±1.12、44.65±1.05、63.12 ±0.67、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、5.85 ±0.06(24 h);2.99±0.09、2.86±0.09、4.70±0.13、3.38±0.07、7.86±0.29、3.54 ±0.03(48 h),ERCC1 mRNA与顺铂IC50的等级相关系数分别为0.89(24 h)、0.94(48 h).结论 ERCC1 mRNA的表达与顺铂作用呈等级相关,ERCC1蛋白表达与顺铂作用无相关,ERCC1 mRNA可应用于临床试验来预测胃癌患者对顺铂的敏感性.
目的 探討6種胃癌細胞株中切除脩複交扠互補基因1(ERCC1)與順鉑作用的關繫.方法 常規培養6種胃癌細胞株；SYBR Green實時定量聚閤酶鏈反應(Real-time PCR)法及Western blot法檢測6種胃癌細胞株的ERCC1 mRNA及蛋白的錶達；不同濃度順鉑作用6、12、24、48 h後,細胞計數試劑盒(CCK-8)法檢測細胞存活率,併計算半數抑製濃度(IC50),Spearman等級相關分彆計算ERCC1 mRNA、蛋白錶達與順鉑作用的相關性.結果 6種胃癌細胞株ERCC1 mRNA的相對錶達量為1.40±0.12、1.35±0.12、3.91 ±0.21、2.92±0.21、4.45 ±0.14、2.26±0.41,ERCC1蛋白錶達灰度分析值為0.58 ±0.04、0.33±0.07、1.07 ±0.13、0.40±0.05、1.38±0.17、0.79 ±0.11,順鉑半數抑製濃度(IC50,mag/L)分彆為36.88±1.29、39.89±1.21、49.72±1.12、44.65±1.05、63.12 ±0.67、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、5.85 ±0.06(24 h);2.99±0.09、2.86±0.09、4.70±0.13、3.38±0.07、7.86±0.29、3.54 ±0.03(48 h),ERCC1 mRNA與順鉑IC50的等級相關繫數分彆為0.89(24 h)、0.94(48 h).結論 ERCC1 mRNA的錶達與順鉑作用呈等級相關,ERCC1蛋白錶達與順鉑作用無相關,ERCC1 mRNA可應用于臨床試驗來預測胃癌患者對順鉑的敏感性.
목적 탐토6충위암세포주중절제수복교차호보기인1(ERCC1)여순박작용적관계.방법 상규배양6충위암세포주；SYBR Green실시정량취합매련반응(Real-time PCR)법급Western blot법검측6충위암세포주적ERCC1 mRNA급단백적표체；불동농도순박작용6、12、24、48 h후,세포계수시제합(CCK-8)법검측세포존활솔,병계산반수억제농도(IC50),Spearman등급상관분별계산ERCC1 mRNA、단백표체여순박작용적상관성.결과 6충위암세포주ERCC1 mRNA적상대표체량위1.40±0.12、1.35±0.12、3.91 ±0.21、2.92±0.21、4.45 ±0.14、2.26±0.41,ERCC1단백표체회도분석치위0.58 ±0.04、0.33±0.07、1.07 ±0.13、0.40±0.05、1.38±0.17、0.79 ±0.11,순박반수억제농도(IC50,mag/L)분별위36.88±1.29、39.89±1.21、49.72±1.12、44.65±1.05、63.12 ±0.67、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、5.85 ±0.06(24 h);2.99±0.09、2.86±0.09、4.70±0.13、3.38±0.07、7.86±0.29、3.54 ±0.03(48 h),ERCC1 mRNA여순박IC50적등급상관계수분별위0.89(24 h)、0.94(48 h).결론 ERCC1 mRNA적표체여순박작용정등급상관,ERCC1단백표체여순박작용무상관,ERCC1 mRNA가응용우림상시험래예측위암환자대순박적민감성.
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 cancer 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 realtime 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 IC50 was calculated respectively.For the correlations,the nonparametric Spearman′s p (r) was calculated.Results Relative expression 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 and0.79±0.11.IC50 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) respectively.Correlation coefficients between ERCC1 mRNA and IC50 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.
