| 王 东,付晓亮,舒 涛,杨增悦,徐 冰.miR-302通过下调靶基因RAB22A抑制膀胱癌进展[J].,2019,19(14):2646-2651 |
| miR-302通过下调靶基因RAB22A抑制膀胱癌进展 |
| MiR-302 Inhibits the Progression of Bladder Cancer by downregulating the Target Gene RAB22A |
| 投稿时间：2018-11-28 修订日期：2018-12-23 |
| DOI：10.13241/j.cnki.pmb.2019.14.009 |
| 中文关键词: 膀胱癌 miR-302 RAB22A 细胞侵袭 |
| 英文关键词: Bladder cancer miR-302 RAB22A Cell invasion |
| 基金项目:国家自然科学基金项目（81700666） |
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| 中文摘要: |
| 摘要 目的：探讨miR-302通过靶向调控RAB22A影响膀胱癌进展的分子机制。方法：采用RT-qPCR检测miR-302在HTB1和RT112膀胱癌细胞系和膀胱内皮细胞系HBdNEC中的表达；以miRNA-NC、miR-302 mimic、miR-302 inhibitor转染细胞，并分为以下几组：NC +control siRNA、miR-302 inhibitor +control siRNA、miR-302 inhibitor +RAB22A siRNA、NC+ vector、miR-302 mimic +vector或miR-302 mimic +RAB22A，再通过MTT实验分析膀胱癌细胞的增殖情况，细胞侵袭实验检测细胞侵袭情况，双荧光素酶报告载体检测分析miR-302靶基因，Western blot检测RAB22A在膀胱癌细胞中的表达。结果：HTB1和RT112细胞中miR-302的表达明显低于HBdNEC细胞(P<0.05)。miR-302高表达抑制膀胱癌细胞的增殖和侵袭；miR-302低表达时，膀胱癌细胞的增殖和侵袭能力上升。生物信息学和双荧光素酶报告结果显示RAB22A为miR-302的靶基因。miR-302过表达后，细胞荧光素酶活性显著下降(P<0.05)，RAB22A表达下调(P<0.05)；miR-302表达沉默后，细胞荧光素酶活性显著上升(P<0.05)，RAB22A表达上调(P<0.01)。拯救实验显示RAB22A表达沉默可逆转miR-302表达沉默时对膀胱癌细胞增殖和侵袭能力上调的影响；而RAB22A过表达可逆转miR-302过表达对膀胱癌细胞增殖和侵袭的抑制。结论：miR-302可通过抑制靶基因RAB22A的表达，抑制膀胱癌细胞的增殖及侵袭。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the molecular mechanism underlying by which miR-302 targeting RAB22A in bladder cancer progression. Methods: The expression of miR-302 in the bladder cell lines HTB1 and RT112 and bladder endothelial cell line HBdNEC was detected by RT-qPCR. miRNA-NC, miR-302 mimic and miR-302 inhibitor were transfected into bladder cancer cells, and divided into the following groups: the NC +control siRNA, miR-302 inhibitor +control siRNA, miR-302 inhibitor +RAB22A siRNA, NC+ vector, miR-302 mimic +vector or miR-302 mimic +RAB22A, respectively. The proliferation was detected by MMT assay, and the invasion was detected by Transwell assay. The dual luciferase reporter assay was performed to verify the target gene of miR-302. And the protein expression of RAB22A in bladder cancer cells was detected by Western blot. Results: The expression of miR-302 in HTB1 and RT112 cells was significantly lower than that in HBdNEC cells. The proliferation and invasion of bladder cancer cells was inhibited by the overexpression of miR-302; While the proliferation and invasion of bladder cancer cells were increased when the expression of miR-302 was silenced. Bioinformatics and dual luciferase reporter results showed that RAB22A was the target gene of miR-302. The activity of cell luciferase and the expression of RAB22A were significantly decreased when miR-302 was overexpressed. However, the activity of cell luciferase and the expression of RAB22A were significantly increased when miR-302 was silenced. The rescue experiments showed that knockdown of RAB22A reversed the increased proliferation and invasion abilities of bladder cancer cells induced by miR-302 inhibitor. Overexpression of RAB22A reversed the inhibition on the proliferation and invasion of bladder cancer cells when miR-302 was overexpressed. Conclusion: miR-302 inhibits the proliferation and invasion of bladder cancer cells by inhibiting the expression of target gene RAB22A. |
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