LncRNA UCA1对卵巢癌细胞侵袭及迁移的影响及机制

摘要

Objective:To study the invasion and migration ability of long chain non-coding RNA UCA1 in ovari-an cancer cells SKOV3. Methods:Constructing eukaryotic expression vector pcDNA/UCA1. pcDNA/UCA1 construct was transfected into SKOV3 cells by lipofectamine2000 for 24h and selected with 500μg/ml G418 for 3 weeks,trans-fected with pcDNA3. 1 empty vector acted as a control. The positive clone was identified by reverse transcription poly-merase chain reaction( RT -PCR ) for UCA 1 RNA expression . Cell invasion and migration ability was assessed by using millicell chamber. MMP2 and MMP9 protein level was detected by Western blotting analysis. Results:We successfully established the SKOV3 cell stable expressing pcDNA/UCA1 transfectant. Exogenous expression of UCA1 in SKOV3 cells enhanced migration and invasion ability of cells. Conclusion:MMP2 and MMP9 protein expression in-creased in SKOV3/pcDNA-UCA1 cells compared with SKOV3/pcDNA3. 1 cells. Which suggested that UCA1 RNA might play some role in SKOV3 cells invasion and progression.%目的：探讨长链非编码RNA UCA1对卵巢癌细胞SKOV3侵袭及迁移能力的影响及可能的作用机制。方法：用脂质体2000将pcDNA/UCA1表达载体及pcDNA3.1空载体转染卵巢癌细胞SKOV3细胞。用G418筛选，建立稳定表达UCA1 RNA的SKOV3/pcDNA-UCA1细胞及转染空载体的SKOV3/pcDNA3.1细胞。RT-PCR方法检测两株细胞中UCA1的表达，鉴定阳性细胞株。Millicell小室检测两株细胞侵袭及迁移能力的变化，蛋白印迹法检测两株细胞MMP2及MMP9蛋白表达的变化。结果：成功构建稳定表达UCA1的SKOV3细胞，表达UCA1 RNA后，SKOV3细胞的侵袭及迁移能力均增加，MMP2及MMP9蛋白表达增加。结论：UCA1 RNA可能通过增加SKOV3细胞中MMP2及MMP9的表达来增强其侵袭及迁移能力，在卵巢癌侵袭及进展中发挥作用。