Starvation microenvironment activates Yes-associated protein 1 to regulate autophagy and promote proliferation of bladder cancer cells

Tinghao LI; Hang TONG; Honghao CAO; Xudong LIU; Junlong ZHU; Zijia QING; Siwen YIN; Weiyang HE

doi:10.16016/j.1000-5404.202106124

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Starvation microenvironment activates Yes-associated protein 1 to regulate autophagy and promote proliferation of bladder cancer cells

Monographic Reports: Basic and clinical researches on bladder cancer | 更新时间：2026-01-09

- Starvation microenvironment activates Yes-associated protein 1 to regulate autophagy and promote proliferation of bladder cancer cells
- Journal of Third Military Medical University Vol. 43, Issue 23, Pages: 2542-2551(2021)
- 作者机构：
  
  1.400016 重庆，重庆医科大学附属第一医院泌尿外科
  2.402460 重庆，重庆市荣昌中医院泌尿外科
  3.402760 重庆，重庆市璧山中医院泌尿外科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81874092)
- DOI：10.16016/j.1000-5404.202106124
  CLC： R341;R730.23;R737.14
- Received：16 June 2021，
  
  Revised：2021-07-18，
  
  Published：15 December 2021
- 稿件说明：
移动端阅览
Tinghao LI, Hang TONG, Honghao CAO, et al. Starvation microenvironment activates Yes-associated protein 1 to regulate autophagy and promote proliferation of bladder cancer cells[J]. Journal of Third Military Medical University, 2021, 43(23): 2542-2551.
DOI：

Tinghao LI, Hang TONG, Honghao CAO, et al. Starvation microenvironment activates Yes-associated protein 1 to regulate autophagy and promote proliferation of bladder cancer cells[J]. Journal of Third Military Medical University, 2021, 43(23): 2542-2551. DOI： 10.16016/j.1000-5404.202106124.

摘要

目的

探究饥饿微环境下膀胱癌细胞恶性增殖能力的变化及其机制。

方法

将膀胱癌T24细胞分为2组，正常组：细胞正常培养，饥饿组：细胞采用不含血清的HBSS处理6 h模拟癌细胞饥饿微环境；分别予以自噬抑制剂氯喹(CQ)或siRNA敲减YAP1基因，观察自噬水平变化以及增殖能力的改变。同时以YAP1及自噬相关基因LC3B表达量构建风险预测模型，分析二者表达量与临床预后的相关性。

结果

CCK-8及克隆形成实验显示饥饿诱导后膀胱癌细胞增殖能力显著增加(

0.01)，且增殖能力的改变是通过增加细胞周期由G

期进入S期，而两组细胞凋亡差异无统计学意义。Western blot及免疫荧光检测P62蛋白降解程度及LC3-Ⅱ/Ⅰ比值，结果显示饥饿可诱导膀胱癌细胞自噬水平上调(

0.05)，而采用CQ阻断自噬流后膀胱癌细胞增殖能力不再受饥饿影响。YAP1蛋白在饥饿诱导后可出现过表达及磷酸化水平的下降(

0.01)，而抑制自噬不影响这一结果。利用siRNA干扰YAP1的表达后膀胱癌细胞自噬水平受到明显抑制，同时增殖能力下降(

0.05)。生物信息学分析发现YAP1在恶性程度高的膀胱癌患者癌组织中表达水平更高，且YAP1与LC3B基因表达水平越高的患者预后更差。

结论

饥饿诱导膀胱癌细胞YAP1的过表达，通过增强自噬水平促进膀胱癌细胞恶性增殖，且YAP1与自噬在膀胱癌发展过程中起到相互协同的促癌作用。

Abstract

Objective

To explore the changes of malignant proliferation of bladder cancer cells under starvation microenvironment and its possible mechanism.

Methods

Bladder cancer T24 cells were divided into 2 groups: the normal group was cultured normally

while the starvation group was treated with serum-free HBSS for 6 h to simulate starvation microenvironment for cancer cells. The changes of autophagy and proliferation ability of cells were observed after treating with autophagy inhibitor CQ or transfection of siRNA of Yes-associated protein 1 (YAP1) gene

respectively. Finally

a risk prediction model was established based on the expression of YAP1 and autophagy-related gene LC3B

and the correlation between their levels and clinical prognosis was subsequently analyzed.

Results

CCK-8 and clone formation assays showed that the proliferation ability of bladder cancer cells was significantly increased after starvation induction (

0.01)

which was caused by promoting cell cycle from G

phase to S phase

while there was no significant difference in apoptosis between the 2 groups. The degradation of autophagy-related marker P62 protein and LC3-Ⅱ/Ⅰ ratio were detected by Western blotting and immunofluorescence assay

and the Results indicated that starvation induced up-regulation of autophagy in bladder cancer cells (

0.05). However

the proliferation of cancer cells was no longer affected by starvation after the autophagy inhibitor CQ blocked the autophagy tide. Overexpression and declined phosphorylation of YAP1 protein were observed after starvation induction (

0.01)

and this result was not affected by inhibition of autophagy. In addition

the autophagy level and the proliferation of cells were significantly inhibited by siRNA interference in YAP1 expression (

0.05). Bioinformatics analysis showed that the level of YAP1 was higher in the cancer tissues of patients with higher malignant grade of bladder cancer

and the patients with higher expression of YAP1 and LC3B genes had worse prognosis.

Conclusion

The overexpression of YAP1 protein induced by starvation promotes the malignant proliferation of bladder cancer cells by enhancing the level of autophagy

and YAP1 and autophagy play a synergistic role in the development of bladder cancer.

关键词

Keywords

references

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