【摘要】 目的:构建长链非编码RNA-UCA1的真核表达载体pcDNA/UCA1,探讨UCA1对膀胱癌细胞BLS-211耐药及转化能力的影响。方法:构建UCA1的真核表达载体pcDNA/UCA1,用脂质体2000将pcDNA/UCA1表达载体及空载体pcDNA3.1转染入膀胱癌细胞BLS-211细胞,用G418筛选,建立稳定表达UCA1RNA的BLS-211/UCA1细胞及转染空载体的BLS-211/pcDNA3.1细胞,用RT-PCR方法检测两株细胞中UCA1及neo基因的表达,鉴定阳性细胞株,通过四甲基偶氮唑蓝(MTT)比色法检测两株细胞对顺铂及丝裂霉素耐药的变化,软琼脂克隆形成实验检测两株细胞克隆形成率的变化。结果:成功构建pcDNA/UCA1真核表达载体,并建立起稳定表达UCA1的BLS-211/UCA1细胞,表达UCA1 RNA后,BLS-211/UCA1细胞对顺铂及丝裂霉素C的耐药性增加,软琼脂克隆形成能力增强。结论:UCA1 RNA增强了膀胱癌细胞BLS-211的耐药能力及转化能力,可能在膀胱癌复发及进展中发挥作用。更多还原

【Abstract】 Objective: To construct the eukaryotic expression vector pcDNA/UCA1 of long chain non- coding RNA- UCA1 and study the transforming ability and drug resistance ability of UCA1 in bladder cancer cells BLS-211. Methods: Constructing eukaryotic expression vector pcDNA / UCA1 of UCA1. The pcDNA / UCA1 construct was transfected into BLS- 2 1 1 cells by lipofectamine 2000 for 24h and selected with 150 μg / ml G418 for 3 weeks,transfected with pcDNA3. 1 empty vector acted as a control. The positive clone was identified by reverse transcription polymerase chain reaction( RT- PCR) for UCA1 and neo gene expression. Cell drug resistance was assessed by using MTT and cell transformation was detected by soft agarose assay. Results: We successfully established the BLS- 211 /UCA1 cell stable expressing pcDNA / UCA1 transfectant. Exogenous expression of UCA1 in BLS- 211 cells enhanced drug resistance and colony forming efficiency in soft agar. Conclusion: UCA1 RNA up- regulation enhanced the drug resistance and transformation ability of BLS- 211 cells,which suggested that UCA1 RNA might play some roles in bladder cancer recurrence and progression.更多还原