Background

As widely reported, propofol can effectively inhibit tumors development. However, little is known about the molecular mechanisms. Here, we proved that propofol regulated miR-340/CDK2 axis to suppress bladder cancer progression in vitro.

Methods

MicroRNA (MiR)-340 expression in 5637 cells was examined using qRT-PCR. Cyclin-dependent kinase2 (CDK2) expression was detected using both qRT-PCR and western blot. The levels of apoptosis-related proteins and cell cycle-related proteins were evaluated using western blot. CCK-8 assay and BrdU assay were conducted to evaluate cell proliferation. Moreover, flow cytometry assay was employed to assess cell cycle and cell apoptosis. Finally, dual luciferase reporter assay was employed to verify the binding relationship between miR-340 and CDK2.

Results

Here we showed that propofol treatment inhibited cell proliferation of 5637 cells but enhanced cell apoptosis. Propofol upregulated miR-340 in a dose and time dependent manner. MiR-340 inhibitor could reverse the effect of propofol on the proliferation and apoptosis of 5637 cells. Next, dual luciferase reporter assay displayed that miR-340 directly bound to the 3′-UTR of CDK2. Finally, inhibition of CDK2 could partly reversed the effect of miR-340 inhibitor on cell proliferation and cell apoptosis of propofol-treated 5637 cells.

Conclusion

In total, our results proved that targeting miR340/CDK2 axis was novel to enhance the anti-tumor effects of propofol in bladder cancer in vitro, and our study provided alternative therapeutic strategies for clinical treatment of bladder cancer.

中文翻译：

---

丙泊酚通过调节 miR-340/CDK2 信号轴抑制膀胱癌细胞增殖并增强其凋亡

背景

正如广泛报道的那样，丙泊酚可以有效地抑制肿瘤的发展。然而，人们对分子机制知之甚少。在这里，我们证明了异丙酚在体外通过调节 miR-340/CDK2 轴来抑制膀胱癌的进展。

方法

使用 qRT-PCR 检查 5637 细胞中的 MicroRNA (MiR)-340 表达。使用 qRT-PCR 和蛋白质印迹检测细胞周期蛋白依赖性激酶 2 (CDK2) 表达。使用蛋白质印迹评估细胞凋亡相关蛋白和细胞周期相关蛋白的水平。进行CCK-8测定和BrdU测定以评估细胞增殖。此外，流式细胞术测定用于评估细胞周期和细胞凋亡。最后，采用双荧光素酶报告基因测定来验证 miR-340 和 CDK2 之间的结合关系。

结果

在这里，我们发现异丙酚处理抑制了 5637 细胞的细胞增殖，但增强了细胞凋亡。丙泊酚以剂量和时间依赖性方式上调 miR-340。MiR-340抑制剂可逆转丙泊酚对5637细胞增殖和凋亡的影响。接下来，双荧光素酶报告基因分析显示 miR-340 直接与 CDK2 的 3'-UTR 结合。最后，抑制 CDK2 可以部分逆转 miR-340 抑制剂对丙泊酚处理的 5637 细胞增殖和细胞凋亡的影响。

结论

总之，我们的研究结果证明了靶向miR340/CDK2轴对于增强丙泊酚在体外对膀胱癌的抗肿瘤作用是新颖的，我们的研究为临床治疗膀胱癌提供了替代治疗策略。