膀胱癌中miR-203的表达及其对吉西他滨化疗敏感性的影响

摘要

目的 探讨microRNA-203(miR-203)在膀胱癌中的表达以及其对吉西他滨化疗敏感性的影响.方法 应用Vita-Blue法检测膀胱癌细胞系对吉西他滨的敏感性差异并测定半数抑制浓度(IC50)值.应用探针法实时荧光定量逆转录-聚合酶链反应(Taqman-qRT-PCR)检测27例膀胱癌组织和癌旁组织以及膀胱癌细胞系5637、Um-Uc-3、J82、SCaBER、 T24、Biu87中miR-203的表达水平.同时构建带有四环素诱导表达系统的miR-203超表达载体(pINDUCER21-EGFP-miR-203), 在吉西他滨化疗耐受的 Um-Uc-3细胞中转染 pINDUCER21-EGFP-miR-203,并用多西环素诱导表达,未诱导表达的为阴性对照,在化疗敏感的T24细胞中分别转染 miR-203 Antagomir 和 Antagomir control,同样使用 Taqman-qRT-PCR 和Vita-Blue方法检测膀胱癌细胞系中miR-203表达水平的变化及对吉西他滨的敏感性.结果 在膀胱癌组织中miR-203的表达显著低于癌旁组织,在膀胱癌细胞系 T24中miR-203的表达水平最高,Um-Uc-3细胞系中表达水平最低(P＜0.01).膀胱癌细胞系T24和Um-Uc-3细胞中对吉西他滨的IC50值分别为(0.46 ±0. 08)ng/ml和(5. 94 ± 0. 53)ng/ml(P＜0.01);pINDUCER21-EGFP-miR-203(D +)可以明显增加Um-Uc-3细胞的化疗敏感性,而miR-203 An-tagomir可以显著抑制T24细胞对吉西他滨的化疗敏感性.结论 miR-203与膀胱癌的发生发展相关,其高表达水平将提高膀胱癌对吉西他滨的化疗敏感性.%Objective To investigate the expression of microRNA-203 (miR-203) in bladder cancer and its effect on chemosensitivity of gemcitabine. Methods The sensitivity of the bladder cancer cell line to gemcitabine was analyzed by Vita-Blue assay and the half-inhibitory concentration (IC50 ) was measured. The expression level of miR-203 was detected in 27 cases of bladder cancer tissues and adjacent tissues by Taqman-qRT-PCR. All the time, the expression levels of miR-203 were also detected in bladder cancer cell lines 5637, Um-Uc-3, J82, SCaBER, T24, Biu87. Construction of miR-203 overexpression vector (pINDUCER21-EGFP-miR-203) with a tetracycline-induced expression system, which was transfected into gemcitabine-resistant Um-Uc-3 cells and was induced with doxycycline(No doxycycline was negative control). Simultaneously, miR-203 Antagomir and Antagomir control were transfected into T24 cells respectively. The sensitivity to gemcitabine and expression levels of miR-203 were also detected using Vita-Blue and Taqman-qRT-PCR in T24 and Um-Uc-3. Results The expression of miR-203 was significantly lower in bladder cancer tissues than in adjacent tissues. The expression level of miR-203 was the highest in bladder cancer cell line T24 and the lowest in Um-Uc-3 cell line (P ＜0. 01). Bladder cancer cell line T24 and Um-Uc-3 to IC50 values of gemcitabine chemotherapy were (0. 46 ±0. 08) ng/ml and (5. 94 ±0. 53) ng/ ml, respectively (P ＜0. 01);pINDUCER21-EGFP-miR-203 (D + ) can remarkably enhance its chemosensitivity in Um-Uc-3, whereas miR-203 Antagomir can also significantly suppress the chemosensitivity of T24 cells to gemcitabine. Conclusion miR-203 is associated with the development and progression of bladder cancer, and its high expression level may improve the chemosensitivity of bladder cancer to gemcitabine.