卵巢癌细胞系RMG-Ⅰ Lewis(y)抗原含量变化对其卡铂耐药性的影响

The Effects of Lewis(y) Antigenic Content on Drug Resistance to Carboplatin in Ovarian Cancer Line RMG-Ⅰ

利用Lewis(y)抗原稳定高表达细胞株RMG-Ⅰ-H,研究细胞表面Lewis(y)抗原含量变化与细胞对卡铂耐药性的关系.利用四甲基偶氮唑盐(MTT)法测定不同浓度卡铂作用后细胞系RMG-Ⅰ-H及其对照组RMG-Ⅰ、RMG-Ⅰ-C的细胞生长抑制率(IR);利用流式细胞仪(FCM)分析细胞凋亡,同时检测药物作用后细胞的凋亡比率;利用荧光染色法和透射电镜进一步观察细胞凋亡状态.结果表明,RMG-Ⅰ-H的半数抑制浓度(IC50)为(58.07±2.42),明显高于对照组RMG-Ⅰ(28.83±3.57)和RMG-Ⅰ-C(25.71±8.24)的IC50(P<0.01),后两者间无统计学差异(P>0.05),流式细胞仪分析证实3组细胞均存在凋亡,经30mg/L、60mg/L卡铂处理后细胞再进行流式细胞仪检测,RMG-Ⅰ-H的相对凋亡率分别为(20.43±0.71)%和(38.11±0.33)%,为对照组的49%～63%,明显低于对照组(P<0.05),2个对照组间比较无统计学差异(P>0.05),荧光染色和透射电镜直接观察了细胞形态变化,每个浓度组RMG-Ⅰ-H的凋亡程度均低于RMG-Ⅰ和RMG-Ⅰ-C.说明细胞表面Lewis(y)抗原含量增加的同时,卵巢癌细胞对卡铂的耐药性增强.

To investigate the relationship between the changes of Lewis （y） antigenic content and the drug resistance of the cells to carboplatin in ovarian cancer lines with RMG- I -H which expresseed higher Lewis（y） antigen on cell surface stably. RMG-I was one kind of epithelial ovarian cancer cell lines. Cell lines RMG- I -C and RMG- I -H were obtained from RMG- I which were transfected with keno-plasmid and α1, 2-fucosyltransferase gene, respectively. The method of methyl thiazolyl tetrazolium （MTT） was used to determine the fractional inhibition ratio （IR） of the three cell lines which were affected by carboplatin in different concentrations. Accordingly, the IC50 of the three cell lines could be calculated. Carboplatin induced apoptosis of three cell lines was observed by flow cytometry （FCM）. In addition, the apoptotic ratios of the three lines that were treated by 30 mg/L and 60 mg/L carboplatin were measured by the FCM at the same time. And then, the apoptosis conditions of the three lines were observed by the methods of fluorescent staining and transmission electron microscope （TEM）. The IC50 ofRMG- I -H was （58.07±2.42）, which was obviously higher than that ofRMG- I （28.83±3.57） and RMG- I -C（25.71±8.24）, all showed a significant statistical difference （P 〈 0.01） and there was no statistical difference between the latter two ones（P 〉 0.05）. FCM analysis confirmed the cell apoptosis in the three lines. The corr-apoptotic ratios of RMG- I -H treated with 30 mg/L and 60 mg/L carboplatin were （20.43 ±0.71）% and （38.11±0.33）%, respectively, which were about 49%-63% of the RMG- I （35.87±3.84）%, （63.37±9.59）% and RMG- I -C（34.80±3.59）%, （60.17±6.64）%, both presented a statistical difference （P 〈 0.05）. But there was no significant statistical difference between the latter two ones （P 〉 0.05）. The apoptotic degree of RMG- I -H was lower than that of the other cell lines in the same carboplatin concentration with the method of inv-fluorescence microscopy and transmission electron microscopy （TEM）. The drug resistance of ovarian cancer cells to carboplatin was stronger when the Lewis（y） antigenic contents on cell surface increased.