| 杨 栋,赵鹏程,马 彦.PlncRNA-1通过吸附miR-141-3p调控雄激素受体表达对前列腺癌细胞增殖的影响[J].中国肿瘤,2020,29(2):154-160. |
| PlncRNA-1通过吸附miR-141-3p调控雄激素受体表达对前列腺癌细胞增殖的影响 |
| PlncRNA-1 regulates Androgen Receptor by Sponging miR-141-3p and Its Effect on Proliferation of Prostate Cancer Cells |
| 中文关键词 修订日期：2019-06-11 |
| DOI：10.11735/j.issn.1004-0242.2020.02.A012 |
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| 中文关键词: 雄激素受体 前列腺癌 miR-141-3p 长链非编码RNA 增殖 |
| 英文关键词:androgen receptor prostate cancer miR-141-3p long non-coding RNA proliferation |
| 基金项目:2018年度河南省医学科技攻关计划项目（2018020506） |
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| 中文摘要: |
| 摘 要：［目的］ 探讨前列腺癌组织长链非编码RNA1（plncRNA-1）对雄激素受体（AR）的调控机制，以及对前列腺癌细胞LNCaP增殖能力的影响。［方法］ 体外培养LNCaP细胞和PC3细胞，利用RT-PCR法和Western blot法检测 plncRNA-1、AR、miR-141-3p在两种细胞中的表达差异。利用荧光原位杂交（FISH）确定plncRNA-1亚细胞定位和核质分离确定plncRNA-1在LNCaP细胞中的定位，并通过RNA pull-down实验验证plncRNA-1和AR是否结合。采用瞬时转染法分别将sh plncRNA-1和阴性对照序列（sh NC）转染至LNCaP细胞中，另外设置空白对照组（Blank）。利用RT-PCR法和Western blot法检测下调plncRNA-1表达后对AR、miR-141-3p的影响。MTT法检测Blank组、sh NC组、sh plncRNA-1组细胞增殖能力的差异。采用双荧光素酶报告基因方法验证plncRNA-1和miR-141-3p，以及miR-141-3p和AR的结合关系。［结果］ LNCaP细胞中plncRNA-1表达量和AR的转录水平高于PC3细胞，miR-141-3p表达量却降低（P<0.05）。FISH和核质分离实验发现plncRNA-1主要定位于细胞质。RNA pull-down实验证实plncRNA-1并不能直接与AR结合。而双荧光素酶基因报告分析显示miR-141-3p既有plncRNA-1的结合位点，也有AR的结合位点。下调plncRNA-1表达后，sh plncRNA-1组LNCaP细胞中plncRNA-1表达量降低，增殖活性也相应降低，但是miR-141-3p的表达量却明显升高，且AR蛋白的表达量也较Blank组和sh NC组细胞明显降低（P<0.05）。［结论］ plncRNA-1通过吸附miR-141-3p调控雄激素受体的表达，从而保护AR的表达和功能活性，下调plncRNA-1表达可降低AR的表达，抑制LNCaP细胞的增殖活性 |
| 英文摘要: |
| Abstract：［Purpose］ To investigate the mechanism of prastate cancer long non-coding RNA1(plncRNA-1) in regulation of androgen receptor(AR) and its effect on the proliferation of prostate cancer cells.［Methods］ The expression of plncRNA-1，AR，miR-141-3p in prostate cancer LNCaP and PC3 cells were detected by qRT-PCR and Western blot. The distribution of plncRNA-1 in LNCaP cells was observed by fluorescence in situ hybridization(FISH). Coimmunoprecipitation of plncRNA-1 and AR was detected by RNA pull-down method. The plncRNA-1 shRNA(sh plncRNA-1 group) and negative sequence(sh NC group) were designed，synthesized and transfected into LNCaP cells. LNCaP cells without any treatment was served as blank group. The plncRNA-1，AR，miR-141-3p levels in blank group，sh NC group and sh plncRNA-1 group were also detected. And the proliferation of LNCaP cells in each group was detected by MTT. The dual luciferase reporter gene system was used to confirm the correlation of miR-141-3p with plncRNA-1 and AR. ［Results］ The plncRNA-1 and AR levels in LNCaP cells were higher than those in PC3 cells，but miR-141-3p expression was lower(P<0.05). PlncRNA-1 was mainly expressed in the cytoplasm of LNCaP cells，and was not significantly enriched in RNAs coimmunoprecipitation with AR. The dual luciferase reporter gene system showed that there would be the binding sites in miR-141-3p with plncRNA-1 and AR mRNA respectively. After down-regulation of plncRNA-1，plncRNA-1 and AR levels in sh plncRNA-1 group were decreased and miR-141-3p level was increased compared to blank group and sh NC group(P<0.05). Moreover，the proliferation activity of LNCaP cells in sh plncRNA-1 group was also inhibited.［Conclusion］ The study shows that plncRNA-1 protects AR expression and its activity. Down-regulation of plncRNA-1 could inhibit the proliferation of prostate cancer cells LNCaP by sponging miR-141-3p.。 |
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