膀胱癌中miR-203的表达及其对吉西他滨化疗敏感性的影响

Expression of miR-203 in bladder carcinoma and its sensitivity to gemcitabine chemotherapy

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(IC_(50))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、SCa BER、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-EGFPmiR-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TaqmanqRT-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IC_(50)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 Antagomir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 gemeitabine. Methods The sensitivity of the bladder cancer cell line to gemcitabine was an- alyzed 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-Ue-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 gemeitabine-resistant Um-Ue-3 cells and was induced with doxyeycline（ No doxyeyeline was negative control）. Simultaneously, miR-203 Antagomir and Antagomir control were transfected into T24 cells respectively. The sensitivity to gemeitabine and expression levels of miR-203 were also detected using Vita-Blue and Taqman-qRT-PCR in T24 and Um-Ue-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 ICso values of gemeitabine 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-Ue-3, whereas miR-203 Antagomir can also significantly suppress the ehemosensitivity 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 ehemosensitivity of bladder cancer to gemcitabine.