lncRNA OSTN-AS1调控前列腺癌细胞生物学行为的机制研究

The mechanism of lncRNA OSTN-AS1 regulating the biological behavior of prostate cancer cells

目的探讨长链非编码RNA OSTN-AS1(lncRNA OSTN-AS1)靶向miR-1207-3p在前列腺癌(PCa)中的作用和分子机制。方法通过RT-qPCR检测OSTN-AS1和微小RNA-1207-3p(miR-1207-3p)在PCa组织中的水平。双荧光素酶报告基因检测验证miR-1207-3p与OSTN-AS1之间的靶点关系。在PCa细胞LNCaP中分别转染si-OSTNAS1(si-OSTN-AS1组)、si-NC(si-NC组)、pcDNA(pcDNA组)、pcDNA-OSTN-AS1(pcDNA-OSTN-AS1组)、miR-NC(miRNC组)、miR-1207-3p模拟物(miR-1207-3p模拟物组)、si-OSTN-AS1+anti-miR-NC(si-OSTN-AS1+anti-miR-NC组)、si-OSTN-AS1+anti-miR-1207-3p(si-OSTN-AS1+anti-miR-1207-3p组)。分别通过CCK-8法、transwell侵袭实验、克隆形成法、划痕愈合实验检测细胞活力、侵袭、克隆和迁移能力。结果PCa组织中OSTN-AS1表达上调(1.00±0.13 vs3.61±0.33)(P<0.05),miR-1207-3p表达下调(1.00±0.09 vs 0.27±0.04)(P<0.05)。miR-1207-3p是OSTN-AS1的靶基因。干扰OSTN-AS1表达后LNCaP细胞活力(0.73±0.06 vs 0.52±0.05)、划痕愈合率(64.87±5.48%vs26.34±2.37%)显著降低(P<0.05),miR-1207-3p水平显著升高(P<0.05),侵袭数(124.11±11.99 vs 56.14±4.59)、克隆形成数(85.99±7.55 vs 38.07±3.54)显著减少(P<0.05)。过表达OSTN-AS1后LNCaP细胞miR-1207-3p水平显著降低(P<0.05)。过表达miR-1207-3p后LNCaP细胞活力(0.76±0.05 vs 0.59±0.05)、划痕愈合率(66.74±4.92%vs 35.44±3.48%)显著降低(P<0.05),侵袭数(125.81±11.93 vs 62.61±5.46)、克隆形成数(88.27±6.29 vs 44.17±4.18)显著减少(P<0.05)。下调miR-1207-3p表达部分逆转了干扰OSTN-AS1表达对LNCaP细胞活力以及侵袭、克隆和迁移能力的影响(P<0.05)。结论干扰OSTN-AS1通过促进miR-1207-3p表达可抑制PCa细胞增殖、迁移和侵袭。

Objective To investigate the function and molecular mechanism of lncRNA OSTN-AS1 targeting miR-1207-3p in prostate cancer(PCa).Methods The levels of OSTN-AS1 and miR-1207-3p in PCa tissues were determined by RTqPCR.The target relationship between OSTN-AS1 and miR-1207-3p was validated by dual-luciferase reporter assay.In PCa cells,LNCaP was transfected with si-OSTN-AS1(si-OSTN-AS1 group),si-NC(si-NC group),pcDNA(pcDNA group),pcDNA-OSTN-AS1(pcDNA-OSTN-AS1 group),miR-NC(miR-NC group),miR-1207-3p mimic(miR-1207-3p mimic group),si-OSTN-AS1+anti-miR-NC(si-OSTN-AS1+anti-miR-NC group),si-OSTN-AS1+anti-miR-1207-3p(si-OSTNAS1+anti-miR-1207-3p group).Cell viability,invasion,cloning and migration capabilities were detected by CCK-8 method,transwell invasion test,clone formation method,and scratch healing test.Results OSTN-AS1 was aberrantly upregulated(1.00±0.13 vs 3.61±0.33)(P<0.05)and miR-1207-3p was aberrantly downregulated(1.00±0.09 vs 0.27±0.04)(P<0.05)in PCa tissues.miR-1207-3p is a direct target of OSTN-AS1.After interfering with OSTN-AS1,the cell viability(0.73±0.06 vs 0.52±0.05)and scratch healing rate(64.87±5.48%vs 26.34±2.37%)of LNCaP cells were aberrantly reduced(P<0.05),miR-1207-3p level was significantly increased(P<0.05),and the number of invasion(124.11±11.99 vs 56.14±4.59)and colony formation(85.99±7.55 vs 38.07±3.54)was significantly reduced(P<0.05).The level of miR-1207-3p in LNCaP cells was significantly reduced after overexpression of OSTN-AS1(P<0.05).After overexpression of miR-1207-3p,the cell viability(0.76±0.05 vs 0.59±0.05)and scratch healing rate(66.74±4.92%vs35.44±3.48%)of LNCaP cells were significantly reduced(P<0.05),and the number of invasion(125.81±11.93 vs62.61±5.46)and colony formation(88.27±6.29 vs 44.17±4.18)was significantly reduced(P<0.05).Downregulation of miR-1207-3p partially reversed the influence of interference with OSTN-AS1 expression on LNCaP cells viability,invasion,cloning and migration capabilities(P<0.05).Conclusion Interference with OSTN-AS1 inhibits the proliferation,migration and invasion of PCa cells by promoting miR-1207-3p expression.