| miR-216a-5p靶向作用于PAK2对膀胱癌细胞增殖和凋亡影响的体外研究 |
| 投稿时间：2017-07-24 修订日期：2017-09-17 点此下载全文 |
| 引用本文：邵焕军,赵振伶,郝丽娜,朱家红,孙峰.miR-216a-5p靶向作用于PAK2对膀胱癌细胞增殖和凋亡影响的体外研究[J].医学研究杂志,2018,47(6):151-155 |
| DOI：
10.11969/j.issn.1673-548X.2018.06.035 |
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| 中文摘要:目的 探讨miR-216a-5p在多种膀胱癌细胞系中表达及调控p21活化蛋白激酶2（PAK2）基因对膀胱癌细胞增殖和凋亡的影响。方法 通过实时荧光定量聚合酶链反应（qRT-PCR）检测miR-216a-5p在正常膀胱细胞系SV-HUC-1及膀胱癌细胞系5637、J82、T24和EJ中的表达，选取其中两种表达miR-216a-5p最少的5637和J82为对象，实验分两组：对照组（转染miR-NC）、实验组（转染miR-216a-5p）。利用qRT-PCR法检测PAK2 mRNA的表达。Western blot法检测PAK2、p-Akt和p-ERK蛋白的表达。Cell Counting Kit-8（CCK-8）实验和克隆形成实验检测膀胱癌细胞活力和增殖能力。流式细胞术检测细胞凋亡情况。结果 miR-216a-5p在膀胱癌细胞系中的表达量均低。实验组PAK2基因mRNA及其蛋自的表达明显降低，PAK2 mRNA在5637细胞中对照组和实验组表达量分别为1.01±0.15和0.40±0.11（P<0.01），在J82细胞中对照组和实验组表达量分别为1.00±0.09和0.56±0.14（P<0.01）。p-Akt和p-ERK蛋白表达显著降低。实验组细胞活力明显被抑制，增殖能力明显降低。实验组细胞凋亡显著增加（P<0.01）。结论 miR-216a-5p在多种膀胱癌细胞系中呈低表达，可在体外靶向抑制PAK2基因，抑制膀胱癌细胞系5637和J82的增殖能力，促进细胞凋亡，可能成为具有膀胱癌生物治疗意义的靶标分子。 |
| 中文关键词:膀胱肿瘤 miR-216a-5p p21活化蛋白激酶2 增殖 凋亡 |
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| Effects of miR-216a-5p Targeting PAK2 on Proliferation and Apoptosis of Bladder Cancer Cells in vitro |
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| Abstract:Objective To investigate the expression of miR-216a-5p in various bladder cancer cell lines and regulation of PAK2 gene on the proliferation and apoptosis of bladder cancer cells. Methods The expression of miR-216a-5p in normal bladder cell line SV-HUC-1 and bladder cancer cell lines (5637, J82, T24 and EJ) was detected by real-time quantitative polymerase chain reaction (qRT-PCR). Two of the least miR-216a-5p-expressed cell lines 5637 and J82 were selected as the experimental object. The experiment was divided into two groups:the control group (transfected with miR-NC) and the experimental group (transfected with miR-216a-5p). The expression of PAK2 mRNA was detected by qRT-PCR. The expression of PAK2, p-Akt and p-ERK protein was detected by Western blot. Cell counting kit-8 (CCK-8) and clone formation assay were used to detect the vitality and proliferation of bladder cancer cells. Flow cytometry was used to detect apoptosis. Results The expression levels of miR-216a-5p in bladder cancer cell lines were lower than normal bladder cells. The expression of PAK2 mRNA and protein was significantly decreased in the experimental group. The expression of PAK2 mRNA in the control and experimental groups were 1.01±0.15 and 0.40±0.11(P<0.01)in 5637 cells. The expression of PAK2 mRNA in the control and experimental groups were 1.00±0.09和0.56±0.14(P<0.01) in J82 cells. The expression of p-Akt and p-ERK protein was significantly decreased. The cell viability of the experimental group was significantly inhibited, and the proliferative ability was significantly decreased. The apoptosis of the experimental group was significantly increased(P<0.01). Conclusion miR-216a-5p is low-expressed in a variety of bladder cancer cell lines. It can inhibit PAK2 gene expression in vitro, inhibit the proliferation of bladder cancer cell lines 5637 and J82, promote cell apoptosis, and may become a target molecule with biotherapeutic significance. |
| keywords:Bladder neoplasms miR-216a-5p PAK2 Proliferation Apoptosis |
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