LncRNA UCA1对卵巢癌细胞侵袭及迁移的影响及机制

Invasion and migration effect and mechanism of long non- coding RNA UCA1 in ovarian cancer cells

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pc DNA/UCA1表达载体及pc DNA3.1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/pc DNA-UCA1细胞及转染空载体的SKOV3/pc DNA3.1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。

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.