饥饿微环境激活YAP1蛋白调控自噬促进膀胱癌细胞的增殖

李庭浩; 童行; 曹洪豪; 刘旭东; 朱俊龙; 秦子家; 尹思文; 何卫阳

doi:10.16016/j.1000-5404.202106124

您当前的位置：

首页 >

文章列表页 >

饥饿微环境激活YAP1蛋白调控自噬促进膀胱癌细胞的增殖

专题报道：膀胱癌基础与临床研究 | 更新时间：2026-01-09

- 饥饿微环境激活YAP1蛋白调控自噬促进膀胱癌细胞的增殖
- Starvation microenvironment activates Yes-associated protein 1 to regulate autophagy and promote proliferation of bladder cancer cells
- 第三军医大学学报 2021年43卷第23期页码：2542-2551
- 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：
  
  [何卫阳，E-mail: weiyang361@aliyun.com HE Weiyang, E-mail: weiyang361@aliyun.com
- 基金信息：
  
  国家自然科学基金面上项目(81874092)
- DOI：10.16016/j.1000-5404.202106124
  中图分类号： R341;R730.23;R737.14
- 收稿：2021-06-16，
  
  修回：2021-07-18，
  
  纸质出版：2021-12-15
- 稿件说明：
移动端阅览
李庭浩, 童行, 曹洪豪, 等. 饥饿微环境激活YAP1蛋白调控自噬促进膀胱癌细胞的增殖[J]. 第三军医大学学报, 2021,43(23):2542-2551.

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.
李庭浩, 童行, 曹洪豪, 等. 饥饿微环境激活YAP1蛋白调控自噬促进膀胱癌细胞的增殖[J]. 第三军医大学学报, 2021,43(23):2542-2551. DOI： 10.16016/j.1000-5404.202106124.

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

BRAY F, FERLAY J, SOERJOMATARAM I, et al . Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J ] . CA Cancer J Clin, 2018, 68(6): 394-424. DOI: 10.3322/caac.21492.

贺海蓉, 李莉, 冯敖梓, 等. 1990-2017年全球膀胱癌发病率和死亡率的研究[J]. 中国循证医学杂志, 2020, 20(11): 1257-1265. DOI: 10.7507/1672-2531.202007095.

HE H R, LI L, FENG A Z, et al . The incidence and mortality of global bladder cancer from 1990 to 2017[J ] . Chin J Evid Based Med, 2020, 20(11): 1257-1265. DOI: 10.7507/1672-2531.202007095.

YIN M, JOSHI M, MEIJER R P, et al . Neoadjuvant chemotherapy for muscle-invasive bladder cancer: a systematic review and two-step meta-analysis[J ] . Oncologist, 2016, 21(6): 708-715. DOI: 10.1634/theoncologist.2015-0440.

LI F P, TENG H L, LIU M D, et al . Prognostic value of immune-related genes in the tumor microenvironment of bladder cancer[J ] . Front Oncol, 2020, 10: 1302. DOI: 10.3389/fonc.2020.01302.

RYBSTEIN M D, BRAVO-SAN PEDRO J M, KROEMER G, et al . The autophagic network and cancer[J ] . Nat Cell Biol, 2018, 20(3): 243-251. DOI: 10.1038/s41556-018-0042-2.

DE DUVE C, WATTIAUX R. Functions of lysosomes[J]. Annu Rev Physiol, 1966, 28(1): 435-492. DOI: 10.1146/annurev.ph.28.030166.002251.

MENG Q, XU J, LIANG C, et al . GPx1 is involved in the induction of protective autophagy in pancreatic cancer cells in response to glucose deprivation[J ] . Cell Death Dis, 2018, 9(12): 1187. DOI: 10.1038/s41419-018-1244-z.

SUN X, SHU Y H, XU M T, et al . ANXA6 suppresses the tumorigenesis of cervical cancer through autophagy induction[J ] . Clin Transl Med, 2020, 10(6): e208. DOI: 10.1002/ctm2.208.

TAO J Y, CALVISI D F, RANGANATHAN S, et al . Activation of β-catenin and Yap1 in human hepatoblastoma and induction of hepatocarcinogenesis in mice[J ] . Gastroenterology, 2014, 147(3): 690-701. DOI: 10.1053/j.gastro.2014.05.004.

FERNANDEZ-L A, SQUATRITO M, NORTHCOTT P, et al . Oncogenic YAP promotes radioresistance and genomic instability in medulloblastoma through IGF2-mediated Akt activation[J ] . Oncogene, 2012, 31(15): 1923-1937. DOI: 10.1038/onc.2011.379.

XU M X, GU M, ZHOU J, et al . Interaction of YAP1 and mTOR promotes bladder cancer progression[J ] . Int J Oncol, 2020, 56(1): 232-242. DOI: 10.3892/ijo.2019.4922.

TONG H, YIN H, HOSSAIN M A, et al . Starvation-induced autophagy promotes the invasion and migration of human bladder cancer cells via TGF-β1/Smad3-mediated epithelial-mesenchymal transition activation[J ] . J Cell Biochem, 2019, 120(4): 5118-5127. DOI: 10.1002/jcb.27788..

ZHANG J X, SONG W, CHEN Z H, et al . Prognostic and predictive value of a microRNA signature in stage Ⅱ colon cancer: a microRNA expression analysis[J ] . Lancet Oncol, 2013, 14(13): 1295-1306. DOI: 10.1016/S1470-2045(13)70491-1.

WANG L Y, HUO M F, CHEN Y, et al . Tumor microenvironment-enabled nanotherapy[J ] . Adv Healthc Mater, 2018, 7(8): e1701156. DOI: 10.1002/adhm.201701156.

SATIJA S, KAUR H, TAMBUWALA M M , et al . Hypoxia-inducible factor (HIF): fuel for cancer progression[J ] . Curr Mol Pharmacol, 2021, 14(3): 321-332. DOI: 10.2174/1874467214666210120154929.

WU S J, KUANG H X, KE J, et al . Metabolic reprogramming induces immune cell dysfunction in the tumor microenvironment of multiple myeloma[J ] . Front Oncol, 2020, 10: 591342. DOI: 10.3389/fonc.2020.591342.

D'SOUZA L, BHATTACHARYA D. Plasma cells: You are what You eat[J]. Immunol Rev, 2019, 288(1): 161-177. DOI: 10.1111/imr.12732.

CHEN Y Q, WEI H J, LIU F, et al . Hyperactivation of mammalian target of rapamycin complex 1 (mTORC1) promotes breast cancer progression through enhancing glucose starvation-induced autophagy and Akt signaling[J ] . J Biol Chem, 2014, 289(2): 1164-1173. DOI: 10.1074/jbc.M113.526335.

FAN Q, YANG L, ZHANG X D, et al . Autophagy promotes metastasis and glycolysis by upregulating MCT1 expression and Wnt/β-catenin signaling pathway activation in hepatocellularcarcinoma cells[J ] . J Exp Clin Cancer Res, 2018, 37(1): 9. DOI: 10.1186/s13046-018-0673-y.

HUANG S, LI Y, SHENG G H, et al . Sirtuin 1 promotes autophagy and proliferation of endometrial cancer cells by reducing acetylation level of LC3[J ] . Cell Biol Int, 2021, 45(5): 1050-1059. DOI: 10.1002/cbin.11549.

PATEL S H, CAMARGO F D, YIMLAMAI D. Hippo signaling in the liver regulates organ size, cell fate, and carcinogenesis[J]. Gastroenterology, 2017, 152(3): 533-545. DOI: 10.1053/j.gastro.2016.10.047.

LEHMANN W, MOSSMANN D, KLEEMANN J, et al . ZEB1 turns into a transcriptional activator by interacting with YAP1 in aggressive cancer types[J ] . Nat Commun, 2016, 7: 10498. DOI: 10.1038/ncomms10498.

SU W J, ZHU S K, CHEN K, et al . Overexpressed WDR3 induces the activation of Hippo pathway by interacting with GATA4 in pancreatic cancer[J ] . J Exp Clin Cancer Res, 2021, 40(1): 88. DOI: 10.1186/s13046-021-01879-w.

LI S, ZHU H, CHEN H D, et al . Glucose promotes epithelial-mesenchymal transitions in bladder cancer by regulating the functions of YAP1 and TAZ[J ] . J Cell Mol Med, 2020, 24(18): 10391-10401. DOI: 10.1111/jcmm.15653.

WANG C Z, YAN G X, DONG D S, et al . LncRNA-A TB promotes autophagy by activating Yes-associated protein and inducing autophagy-related protein 5 expression in hepatocellular carcinoma[J ] . World J Gastroenterol, 2019, 25(35): 5310-5322. DOI: 10.3748/wjg.v25.i35.5310.

PAVEL M, RENNA M, PARK S J, et al . Contact inhibition controls cell survival and proliferation via YAP/TAZ-autophagy axis[J ] . Nat Commun, 2018, 9(1): 2961. DOI: 10.1038/s41467-018-05388-x..

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

8-自噬相关lncRNA模型在预测膀胱癌患者预后中的作用

肿瘤微环境中M2巨噬细胞调控CLDN10通路促进膀胱癌细胞的上皮-间质转化

miR-124在膀胱癌中的表达及其临床意义

尿苷三磷酸增加细胞内Ca2+信号诱导自噬抑制胃癌细胞的增殖

硅橡胶硬度通过自噬调控成纤维细胞增殖及α-SMA、CollagenⅠ的表达