| 陈达书,蒋倩颖.长链非编码 RNA linc 00261通过竞争性结合微小 RNA-182调控 RND3基因参与卵巢癌癌细胞的生物学行为研究[J].安徽医药,2021,25(5):903-909. |
| 长链非编码 RNA linc 00261通过竞争性结合微小 RNA-182调控 RND3基因参与卵巢癌癌细胞的生物学行为研究 |
| Long chain noncoding RNA linc 00261 participates in the biological behavior of ovarian can? cer cells by regulating RND3 gene through competitive binding to miR-182 |
| |
| DOI：10.3969/j.issn.1009-6469.2021.05.014 |
| 中文关键词: 卵巢肿瘤 linc 00261 微小 RNA-182 RND3 竞争性结合 凋亡 |
| 英文关键词: Ovarian neoplasms Linc 00261 miR-182 RND3 Competitive binding Apoptosis |
| 基金项目:无锡市卫生和计划生育委员科研项目( FYTG201606) |
|
| 摘要点击次数: 2100 |
| 全文下载次数: 870 |
| 中文摘要: |
| 目的探讨长链非编码 RNA linc 00261通过竞争性结合微小 RNA-182(miR-182)调控 RND3基因参与卵巢癌癌细胞生物学行为研究及其作用机制。方法选取人卵巢表面上皮细胞系 IOSE80和 4株卵巢癌细胞系 SKOV3、HO8910、A2780和 OVCAR。实时荧光定量聚合酶链式反应( qRT-PCR)检测 linc 00261、miR-182和 RND3 mRNA的表达；双荧光素酶实验验证 linc 00261和 miR-182结合， miR-182靶向调控 RND3；RIP验证 linc 00261和 AGO2结合； RNA pull down结果证明 linc 00261和 miR182结合； CCK8检测细胞的生长能力； Transwell实验检测细胞迁移侵袭能力；细胞流式术检测细胞凋亡率；蛋白质免疫印迹(Western Blot，WB)检测 RND3和凋亡相关因子 Bax和 Bcl-2蛋白的表达。结果与人卵巢表面上皮细胞株 IOSE80(1.00±0.11)相比， 4株卵巢癌细胞 SKOV3(0.25±0.04)、 HO8910(0.71±0.08)、 A2780(0.53±0.07)和 OVCAR(0.36±0.05)中 linc 00261的表达显著降低( P < 0.05)，而 miR-182的表达在 SKOV3(3.46±0.42)、 HO8910(2.07±0.21)、 A2780(2.79±0.31)和 OVCAR(2.98±0.32)中均显著增加(均 P < 0.05)，其中 SKOV3中 linc 00261的表达最低。双荧光素酶实验验证 linc 00261能与 miR-182结合， RIP实验验证 AGO2蛋白与 linc 00261结合， RNA pull down实验证明 linc 00261与 miR-182特异性结合，且 miR-182能够靶向调控 RND3的表达。过表达 linc 00261或沉默 miR-182可以抑制卵巢癌细胞的增殖、迁移和侵袭能力，并促进其凋亡，凋亡相关因子 Bax表达上调，而抗凋亡相关因子 Bcl-2表达下降，而过表达 miR-182或沉默 RND3可以逆转 linc 00261过表达对卵巢癌细胞生物学活性的抑制作用。结论长链非编码 RNA linc 00261通过竞争性结合 miR-182，从而上调 RND3基因，进而抑制卵巢癌癌细胞增殖、迁移和侵袭，并促进其凋亡。 |
| 英文摘要: |
| Objective To investigate the role of long-chain noncoding RNA linc 00261 in the regulation of RND3 gene by competitive binding to miR-182 in participating the biological behavior of ovarian cancer cells.Methods In this study, human ovarian epithelial cell lines IOSE80 and four ovarian cancer cell lines SKOV3, HO8910, A2780 and OVCAR were selected. Detection of mRNA expression of linc 00261, miR-182 and RND3 by qRT-PCR. The binding of linc 00261 and miR-182 was confirmed by double luciferase experiment, and miR-182 targeted to regulate RND3. RIP verifies the combination of linc 00261 and AGO2. The results of RNA pulldown showed that linc 00261 combined with miR-182. CCK8 test cell growth ability. Transwell test was used to detect the ability of cellmigration and invasion. Flow cytometry was used to detect the apoptosis rate. WB was used to detect the expression of RND3 and apoptosis related factors Bax and Bcl-2.Results In ovarian cancer cells, Linc 00261 and Rnd3 were low expression[SKOV3(0.25±0.04),HO8910(0.71±0.08),A2780(0.53±0.07) and OVCAR(0.36±0.05) vs.IOSE80(1.00±0.11)], while mir-182 was high expression[SKOV3 (3.46±0.42),HO8910(2.07±0.21),A2780(2.79±0.31) and OVCAR(2.98±0.32)](P < 0.05). Linc 00261 can bind to miR-182 by double luciferase assay, and AGO2 can bind to linc 00261 by RIP assay, and linc 00261 can specifically bind to miR-182 by RNA pull down assay, and miR-182 can target and regulate the expression of RND3. Overexpression of linc 00261 or silence of miR-182 can inhibit the proliferation, migration and invasion of ovarian cancer cells, and promote their apoptosis. The expression of Bax is up-regulated, while Bcl-2 is down regulated, but overexpression of miR-182 or silencing of RND3 could reverse the inhibition of biological activity of ovari an cancer cells by overexpression of Linc 00261.Conclusion Long-chain noncoding RNA linc 00261can up regulate RND3 gene by com petitive bindingmi R-182,and then inhibit the proliferation,migration andinvasion of ovarian cancer cells,and promote their apoptosis. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
