MicroRNA-153在膀胱癌组织中的表达及对膀胱癌细胞生物学特性的影响

MicroRNA-153 expression in bladder cancer tissues and the impact on the biological characteristics of bladder cancer cells

目的了解MicroRNA-153(miR-153)在膀胱癌组织中的表达及对膀胱癌细胞生物学特性的影响。方法收集经病理确诊为膀胱癌进行外科手术切除的25例患者的膀胱癌组织及距离膀胱癌病灶5 cm以上的癌旁组织,通过实时定量聚合酶链反应(Real time PCR)检测miR-153在25对膀胱癌及瘤旁组织中的表达,分析miR-153基因表达与肿瘤分类(肌层浸润型与非肌层浸润型)及病理分级的关系。采用Annexin V/PI双染法和MTT实验绘制细胞生长曲线,采用生物信息学软件及荧光素酶实验验证miR-153是否存在直接作用。结果与癌旁组织相比,miR-153在膀胱癌组织中的相对表达量分别为1.04±0.07和0.14±0.03。膀胱癌组织显著低于癌旁组织,差异有统计学意义(P<0.05);miR-153在肌层浸润组中的表达为非肌层浸润组的(2.48±0.32)倍,其表达水平随着病理分级升高而升高(P<0.05);miR-153成熟体的膀胱癌EJ细胞中RAS蛋白的表达水平较对照组显著降低,且具有浓度依赖性。结论miR-153在膀胱癌组织中的相对表达量明显升高,且其表达水平随着病理分级升高而升高。由于受到脂质体毒性的影响,肿瘤细胞的增殖受到抑制,anti-miRl53组的活性细胞数明显下降。anti-miRl53特异性地抑制了膀胱癌细胞的增殖。特异性anti-miRl53的转染使膀胱癌细胞发生了包括凋亡和坏死在内的肿瘤细胞的死亡,提示有可能参与了膀胱癌细胞的增殖和凋亡过程,并可作为膀胱癌的潜在临床诊断标志物。

Objective To investigate the expression and MicroRNA- 153 in bladder carcinoma and bladder cancer cells biological characteristics. Methods Pathologically diagnosed as bladder cancer surgical resection tissue of 25 patients with the bladder carcinoma and bladder cancer lesion 5 cm above the adjacent tissues were analyzed by real- time quantitative polymerase chain reaction（ Real time PCR） to detect miR- 153 in bladder tumor adjacent tissues and 25 pairs. We also analyzed the relationship between miR- 153 gene expression and tumor classification（ muscle invasive and non- muscle invasive type） and pathological grading. Using Annexin V / PI double staining and MTT assay and cell growth curve,using bioinformatics software and luciferase experiments miR- 153 to investigate whether there was a direct effect. Results Compared with the adjacent tissues,the relative expression of miR- 153 in bladder cancer tissues were 1. 04 ± 0. 07 and 0. 14 ± 0. 03. Bladder cancer tissues was significantly lower than adjacent tissues,the difference was statistically significant（ P〈0. 05）. The expression of miR- 153 in non- muscle- invasive group（ 2. 48 ± 0. 32） times the muscle invasive group,the expression level with temperature increased with tumor grade（ P〈0. 05）. The expression level of bladder cancer EJ cells mature form of miR- 153 in the RAS protein than the control group significantly decreased in a dose- dependent manner. Conclusion miR- 153 in the relative expression in bladder cancer tissue volume increased significantly,and its expression level increased with pathologic grading and rise. Under the influence of liposome toxicity,the proliferation of tumor cells is affected,the activity of anti- miRl53 group cells was significantly lower. Anti- miRl53 specificity to inhibit bladder cancer cell proliferation.Specific anti- miRl53 transfection of bladder cancer cells have taken place in the death of tumor cells,including apoptosis and necrosis,prompt likely to be involved in the process of bladder cancer cell proliferation and apoptosis,and can be used as potential clinical diagnosis of bladder cancer markers.