Department of Urology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.
Center for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, China.
PeerJ. 2022 Jan 26;10:e12724. doi: 10.7717/peerj.12724. eCollection 2022.
To investigate the mechanism of miR-148a-3p regulating the proliferation and migration of bladder tumor cells.
We conducted a preliminary study to detect the relative expression of miR-148a-3p in bladder cancer and para-cancerous tissue samples. Three bladder tumor cell lines, T24, 5,637 and UM-UC-3, were selected. The expression levels of miR-148a-3p were artificially regulated with miR-148a-3p mimics and the miR-148a-3p inhibitor. The relative expression levels of miR-148a-3p in the samples of each cell line were determined. Cell Counting Kit-8 (CCK-8) was used to detect cell proliferation, while the effect of the miR-148a-3p mimics and inhibitor on tumor cell migration was detected by wound healing assay. Flow cytometry assay was carried out to explore the effect of miR-148a-3p on cell apoptosis. Dual-luciferase reporter assay was performed in order to verify miR-148a-3p's target gene. The expressions of ROCK-1 and Bcl-2 were analyzed by western blot.
The relative expression of miR-148a-3p in tumor and adjacent tissues was assessed with qRT-PCR ( < 0.05) and found to be significantly lower in the tumor tissues than the adjacent tissues. The data obtained from the CCK-8 and wound healing assay showed that intracellular transfection of miR-148a-3p mimics could inhibit cell proliferation and migration, while the miR-148a-3p inhibitor promoted them. Overexpression of miR-148a-3p promoted cell apoptosis in the T24 and 5,637 cell lines. The dual-luciferase reporter assay verified that ROCK-1 is a direct target of miR-148a-3p. Western blot showed that miR-148a-3p overexpression downregulated the expression of ROCK-1 and Bcl-2, while miR-148a-3p knockdown upregulated the expression of ROCK-1 and Bcl-2.
We confirmed that miR-148a-3p was significantly decreased in bladder cancer cells. miR-148a-3p overexpression inhibited bladder cancer cell proliferation and migration, whereas miR-148a-3p knockdown promoted bladder cancer cell proliferation and migration. Moreover, we found that ROCK-1 was a downstream target of miR-148a-3p. We also found that miR-148a-3p induced cell apoptosis by regulating the expression of Bcl-2. However, the deeper mechanism of this regulatory relationship needs further study.
探讨 miR-148a-3p 调控膀胱肿瘤细胞增殖和迁移的机制。
我们进行了一项初步研究，以检测膀胱癌和癌旁组织样本中 miR-148a-3p 的相对表达水平。选择了三个膀胱肿瘤细胞系，T24、5,637 和 UM-UC-3。用 miR-148a-3p 模拟物和 miR-148a-3p 抑制剂人工调节 miR-148a-3p 的表达水平。检测每个细胞系样本中 miR-148a-3p 的相对表达水平。细胞计数试剂盒（CCK-8）用于检测细胞增殖，而划痕愈合试验检测 miR-148a-3p 模拟物和抑制剂对肿瘤细胞迁移的影响。流式细胞术检测 miR-148a-3p 对细胞凋亡的影响。双荧光素酶报告实验验证 miR-148a-3p 的靶基因。Western blot 分析 ROCK-1 和 Bcl-2 的表达。
qRT-PCR 评估了肿瘤组织和癌旁组织中 miR-148a-3p 的相对表达（<0.05），结果显示肿瘤组织中的 miR-148a-3p 表达明显低于癌旁组织。CCK-8 和划痕愈合试验的数据表明，miR-148a-3p 模拟物的细胞内转染可抑制细胞增殖和迁移，而 miR-148a-3p 抑制剂则促进其增殖和迁移。miR-148a-3p 的过表达促进了 T24 和 5,637 细胞系中的细胞凋亡。双荧光素酶报告实验验证了 ROCK-1 是 miR-148a-3p 的直接靶标。Western blot 显示 miR-148a-3p 过表达下调了 ROCK-1 和 Bcl-2 的表达，而 miR-148a-3p 敲低则上调了 ROCK-1 和 Bcl-2 的表达。
我们证实 miR-148a-3p 在膀胱癌细胞中显著降低。miR-148a-3p 的过表达抑制膀胱癌细胞的增殖和迁移，而 miR-148a-3p 的敲低则促进膀胱癌细胞的增殖和迁移。此外，我们发现 ROCK-1 是 miR-148a-3p 的下游靶标。我们还发现 miR-148a-3p 通过调节 Bcl-2 的表达诱导细胞凋亡。然而，这种调控关系的更深层次机制需要进一步研究。
