摘要目的 探讨长链非编码RNA (long non-coding RNA,lncRNA)尿路上皮癌相关1(urothelial carcinoma associated 1,UCA1)基因和miR-34b在膀胱癌细胞和膀胱癌组织中的表达及临床相关性.方法 2011年1月至2012年10月采用实时荧光定量聚合酶链反应(PCR)检测UCA1基因和miR-34b在人膀胱上皮永生化细胞株SV-HUC-1及5种膀胱癌细胞株(T24、BIU-87、EJ、T24-MMC、T24-ADM)中的表达情况.同时收集56例膀胱癌患者的根治性膀胱切除术后病理标本,包括膀胱癌组织及其癌旁正常组织(病理诊断为正常膀胱黏膜组织).男41例,女15例.年龄52～78岁,平均(68.4±7.5)岁,其中≥65岁43例,＜65岁13例.非肌层浸润性膀胱癌18例,肌层浸润性膀胱癌38例.病理类型均为乳头状尿路上皮癌,其中低级别22例,高级别34例.肿瘤初发12例,复发44例.采用实时荧光定量PCR检测UCA1基因和miR-34b的表达情况,分析两者与膀胱癌临床病理特征的关系.结果 UCA1基因在人膀胱上皮永生化细胞株SV-HUC-1和人膀胱癌T24、BIU-87、EJ、T24-MMC、T24-ADM细胞株中的相对表达量分别为(0.0675 ±0.0133)、(0.2934 ±0.0531)、(0.4246±0.0650)、(0.4206±0.0826)、(0.6472±0.0875)和(0.7165±0.1032).与SV-HUC-1细胞株比较,UCA1基因在5种人膀胱癌细胞株中的表达均上调;且UCA1基因在膀胱癌耐药细胞株T24-MMC和T24-ADM中的表达量明显高于非耐药膀胱癌细胞株T24,差异有统计学意义(P＜0.05).miR-34b在SV-HUC-1细胞株和人膀胱癌T24、BIU-87、EJ、T24-MMC、T24-ADM细胞株中的表达量分别为(0.6384±0.1083)、(0.1600±0.0455)、(0.1720±0.0658)、(0.1150±0.0352)、(0.0576±0.0087)和(0.0510±0.0125).与SV-HUC-1细胞株比较,miR-34b在5种人膀胱癌细胞株中的表达均下调;且miR-34b在膀胱癌耐药细胞株T24-MMC和T24-ADM中的表达量明显少于非耐药膀胱癌细胞株T24,差异有统计学意义(P＜0.05).膀胱癌组织和癌旁正常组织中UCA1基因的相对表达量分别为(0.4225±0.0714)和(0.0532±0.0192),miR-34b的相对表达量分别为(0.0340±0.0134)和(0.5643±0.0616),差异有统计学意义(P＜0.05).在膀胱癌组织中,UCA1基因与miR-34b的表达呈负相关(r=-0.54,p＜0.05).UCA1基因表达上调,miR-34b表达下调,且与肿瘤的分级、有无肌层浸润和有无复发密切相关.UCA1基因高表达同时miR-34b低表达组[UCA1(+)/miR-34b(-)]患者的3年总生存率为27.6％,低于非UCA1(+)/miR-34b(-)组患者的73.7％,差异有统计学意义(P＜0.05).结论 UCA1基因和miR-34b与膀胱癌的预后密切相关,且两者的表达量具有相关性.

abstractsObjective To study the expression of long non-coding RNA (lncRNA)-urothelial carcinoma associated 1 (UCA1) and miR-34b in bladder cancer and its correlation to the clinicopathologic features of bladder cancer.Methods Between January 2011 and October 2012,the expression of UCA1 and miR-34b in 5 bladder cancer cell lines (T24,BIU-87,EJ,T24-MMC,T24-ADM) and 1 normal bladder cell lines (SV-HUC-1) were measured by real-time reverse transcription-polymerase chain reaction (RTPCR).Meanwhile,the 56 bladder cancer specimens and paraneoplastic normal bladder tissues,which diagnosed by pathology were collected from bladder cancer patients undergoing radical resection of bladder.Among them,41 cases were male and 15 cases were female.The mean age was (68.4 ± 7.5)years old,range 52 to 78 years.43 cases were older than 65 years old,and 13 cases were less than 65 years old.The pathological classification included non muscle-invasive bladder cancer (NMIBC) 18 cases,muscle-invasive bladder cancer 38 cases;low grade papillary urothelial carcinoma 22 cases,high grade papillary urothelial carcinoma 34 cases;12 cases were primary lesion,the other 44 cases were diagnosed as tumor recurrence.Real-time RT-PCR was performed to analyze the expression of UCA1 and miR-34b.Results The relative expression levels of UCA1 in the normal bladder cell lines (SV-HUC-1) and 5 bladder cancer cell lines (T24,BIU-87,EJ,T24-MMC and T24-ADM) were (0.0675 ± 0.0133),(0.2934 ± 0.0531),(0.4246 ± 0.0650),(0.4206 ± 0.0826),(0.6472 ± 0.0875) and (0.7165 ± 0.1032),respectively (P ＜ 0.05).Moreover,the expression levels of UCA1 were up-regulated in 2 drug resistant bladder cancer cells lines T24-MMC (0.6472 ± 0.0875)and T24-ADM (0.7165 ± 0.1032),as compared with the T24 bladder cancer lines (0.2934 ± 0.0531),respectively (P ＜ 0.05).However,the expression levels of miR-34b in 5 bladder cancer cell lines [T24 (0.1600 ± 0.0455),BIU-87 (0.1720 ± 0.0658),EJ (0.1150 ± 0.0352),T24-MMC(0.0576 ± 0.0087),T24-ADM (0.0510 ± 0.0125)] were decreased (P ＜ 0.05),as compared with normal bladder cell lines SV-HUC-1 (0.6384 ± 0.1083).Moreover,the expression levels of miR-34b were down-regulated in 2 drug resistant bladder cancer cells lines T24-MMC (0.0576 ± 0.0087) and T24-ADM(0.0510 ± 0.0125),as compared with the T24 bladder cancer lines T24 (0.1600 ± 0.0455),respectively (P ＜ 0.05).The relative expression levels of UCA1 and miR-34b in bladder cancer tissues and paraneoplastic normal bladder tissues were (0.4225 ± 0.0714) vs.(0.0532 ± 0.0192) and (0.0340 ± 0.0134)vs.(0.5643 ±0.0616),respectively (P ＜0.05).Statistical correlation analysis showed that UCA1 to be significantly negative correlated with miR-34b in bladder cancer specimens(r =-0.54,P ＜ 0.05).The high level of UCA1 and low level of miR-34b were significantly correlated with tumor malignant grade,invasiveness and recurrence.The 3-year overall survival rate (OS) in UCA1 (+)/miR-34b(-) group (27.6％) were significantly worse compared with non UCA1 (+)/miR-34b (-) group (73.7％).Conclusion High expression of UCA1 and low expression of miR-34b were associated with the occurrence and development of bladder cancer.