LRRC59 promotes proliferation, invasion and inhibition of apoptosis in bladder urothelial carcinoma cells

Peng ZHAGN; Peihe LIANG

doi:10.16016/j.2097-0927.202212031

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LRRC59 promotes proliferation, invasion and inhibition of apoptosis in bladder urothelial carcinoma cells

Basic Medicine | 更新时间：2026-01-09

- LRRC59 promotes proliferation, invasion and inhibition of apoptosis in bladder urothelial carcinoma cells
- Journal of Army Medical University Vol. 45, Issue 14, Pages: 1526-1538(2023)
- 作者机构：
  
  400010 重庆，重庆医科大学附属第二医院泌尿外科
- 作者简介：
- 基金信息：
  
  the Project for High-Level Reserved Young and Middle-Aged Medical Talents of Chongqing(第四批);the Basic Science and Frontier Technology Research Project of Chongqing(CSTC2017jcyjAX0435);the Entrepreneurship and Innovation Supporting Program for Overseas Returnees of Chongqing(CX2019146)
- DOI：10.16016/j.2097-0927.202212031
  CLC： R341;R730.23;R737.14
- Received：05 December 2022，
  
  Revised：2023-02-01，
  
  Published：30 July 2023
- 稿件说明：
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Peng ZHAGN, Peihe LIANG. LRRC59 promotes proliferation, invasion and inhibition of apoptosis in bladder urothelial carcinoma cells[J]. Journal of Army Medical University, 2023, 45(14): 1526-1538.
DOI：

Peng ZHAGN, Peihe LIANG. LRRC59 promotes proliferation, invasion and inhibition of apoptosis in bladder urothelial carcinoma cells[J]. Journal of Army Medical University, 2023, 45(14): 1526-1538. DOI： 10.16016/j.2097-0927.202212031.

摘要

目的

探讨富含亮氨酸重复序列蛋白59(leucine-rich repeat-containing protein 59，LRRC59)在膀胱尿路上皮癌中的表达及其对肿瘤细胞增殖、凋亡及侵袭能力的影响。

方法

收集重庆医科大学附属第二医院泌尿外科2020年10月至2021年9月膀胱尿路上皮癌患者组织标本10例，使用免疫组化技术分析膀胱癌组织及癌旁组织中LRRC59表达情况。用在线数据库(TIMER2.0、GEPIA2、Linked Omics)分析泛癌、膀胱癌及正常膀胱中LRRC59的表达情况并分析其预后，通过分析膀胱癌中具有关联性的基因进行通路富集分析。RT-qPCR及Western blot检测膀胱尿路上皮癌细胞系T24、5637、J82和正常尿路上皮细胞系SV-HUC-1中LRRC59表达，以SV-HUC-1细胞作为对照。选择T24及5637细胞系，通过质粒转染shRNA下调LRRC59表达，以shRNA-NC作为对照组，shRNA-1、shRNA-2、shRNA-3作为实验组。CCK8和克隆形成实验检测细胞增殖能力，Transwell和细胞划痕实验检测细胞迁移、侵袭能力，AnnexinV-FITC、碘化丙啶(PI)染色后流式细胞术检测细胞凋亡和周期，Western blot检测上皮间质转化、凋亡及p53通路相关蛋白的表达。

结果

免疫组化结果显示LRRC59在膀胱尿路上皮癌组织中的表达高于癌旁组织(

0.05)；数据库检索显示LRRC59在膀胱癌患者中表达高于正常者(

0.05)，且患者中高表达者预后更差(

0.01)。通路富集分析提示

LRRC

59相关基因主要富集在内质网蛋白质加工、细胞周期与DNA复制；敲低LRRC59的表达能抑制膀胱癌细胞的增殖，细胞发生G1期阻滞，凋亡率增加，促凋亡蛋白Bax表达增高，抑凋亡蛋白Bcl-2表达下降；细胞迁移及侵袭能力下降。上皮-间质转化相关分子检测提示Snail、Vimentin蛋白表达下降，而E-cadherin蛋白表达增高(

0.05)。p53通路蛋白检测提示p53、p21蛋白表达升高，而Cyclin D1蛋白表达下降(

0.05)。

结论

LRRC59的表达促进膀胱尿路上皮癌细胞增殖、迁移和侵袭；LRRC59可能是膀胱尿路上皮癌发生发展的关键生物标志物。

Abstract

Objective

To investigate the expression of leucine-rich repeat-containing protein 59 (LRRC59) in bladder urothelial carcinoma and its effect on proliferation

apoptosis and invasion of tumor cells.

Methods

Ten tissue samples were collected from the patients with bladder urothelial carcinoma hospitalized in our hospital from Oct. 2020 to Sep. 2021

and then the expression of LRRC59 in the tumor and adjacent non-cancerous tissues was measured with immunohistochemical assay. The expression of LRRC59 in pan-carcinoma

bladder cancer and normal bladder was also analyzed in Online databases TIMER

GEPIA and LinkedOmics

and its relationship with prognosis of these subjected patients was analyzed. Then pathway enrichment analysis was performed by analyzing the genes related to bladder cancer. LRRC59 expression in bladder urothelial carcinoma cell lines T24

5637 and J82 and human bladder epithelial cell line SV-HUC-1 (as control) was detected by RT-qPCR and Western blotting. After the expression of LRRC59 in T24 and 5637 cells were knocked down by sh-RNA techniques (shRNA-1

shRNA-2 and shRNA-3 for knockdown

and shRNA-NC as blank control). Then cell proliferation was detected by CCK8 assay and colony formation assay

migration and invasion were observed by Transwell assay and cell scratch assay

and cell apoptosis and cycle were measured by flow cytometry after AnnexinV-FITC and Propidium Iodide (PI) staining. The expression of epithelial-mesenchymal transition (EMT) related proteins

apoptosis-related proteins and p53 pathway-related proteins were detected by Western blotting.

Results

The expression of LRRC59 was higher in bladder urothelial carcinoma tissues than adjacent non-cancerous tissues (

0.05). Database analysis showed that LRRC59 expression was higher in the bladder cancer tissue than the normal tissues (

0.01)

and the patients with high LRRC59 expression had a worse prognosis (

0.01). Pathway enrichment analysis suggested that LRRC59-related genes were mainly enriched in protein processing in endoplasmic reticulum

cell cycle and DNA replication. Knockdown of LRRC59 inhibited cell proliferation

arrested cell cycle at G1 phase and improved cell apoptosis

and also promoted the expression of Bax while decreased that of Bcl-2. And

cell migration and invasion abilities were decreased

as well as the expression levels of Snail

vimentin and cyclin D1 were decreased (

0.05)

while those of E-cadherin

p53 and p21 protein were enhanced (

0.05).

Conclusion

LRRC59 promotes the proliferation

migration and invasion of bladder urothelial carcinoma cells

and it may be a key biomarker for development and progression of bladder urothelial carcinoma.

关键词

Keywords

references

SUNG H, FERLAY J, SIEGEL R L, et al . Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J ] . CA Cancer J Clin, 2021, 71(3): 209-249. DOI: 10.3322/caac.21660.

ZHENG R S, ZHANG S W, ZENG H M, et al . Cancer incidence and mortality in China, 2016[J ] . J Natl Cancer Cent, 2022, 2(1): 1-9. DOI: 10.1016/j.jncc.2022.02.002.

TEOH J Y C, HUANG J J, KO W Y K, et al . Global trends of bladder cancer incidence and mortality, and their associations with tobacco use and gross domestic product per capita[J ] . Eur Urol, 2020, 78(6): 893-906. DOI: 10.1016/j.eururo.2020.09.006.

SKJERPEN C S, WESCHE J, OLSNES S. Identification of ribosome-binding protein p34 as an intracellular protein that binds acidic fibroblast growth factor[J]. J Biol Chem, 2002, 277(26): 23864-23871. DOI: 10.1074/jbc.M112193200.

BLENSKI M, KEHLENBACH R H. Targeting of LRRC59 to the endoplasmic reticulum and the inner nuclear membrane[J]. Int J Mol Sci, 2019, 20(2): 334. DOI: 10.3390/ijms20020334.

LUO J H, MICHALOPOULOS G, YU Y P. Identification of recurrent fusion genes across multiple cancer types[J]. Sci Rep, 2019, 9(1): 1074. DOI: 10.1038/s41598-019-38550-6.

YU Y P, DING Y, CHEN Z H, et al . Novel fusion transcripts associate with progressive prostate cancer[J ] . Am J Pathol, 2014, 184(10): 2840-2849. DOI: 10.1038/S41598-019-38550-6.

HUANG L P, SAVOLY D, SIDI A A, et al . CIP2A protein expression in high-grade, high-stage bladder cancer[J ] . Cancer Med, 2012, 1(1): 76-81. DOI: 10.1002/cam4.15.

PANG X, FU X, CHEN S, et al . Overexpression of CIP2A promotes bladder cancer progression by regulating EMT[J ] . Clin Transl Oncol, 2016, 18(3): 289-295. DOI: 10.1007/s12094-015-1366-z.

PALLAI R, BHASKAR A, BARNETT-BERNODAT N, et al . Leucine-rich repeat-containing protein 59 mediates nuclear import of cancerous inhibitor of PP2A in prostate cancer cells[J ] . Tumour Biol, 2015, 36(8): 6383-6390. DOI: 10.1007/s13277-015-3326-1.

PALLAI R, BHASKAR A, BARNETT-BERNODAT N, et al . Cancerous inhibitor of protein p hosphatase 2A promotes premature chromosome segregation and aneuploidy in prostate cancer cells through association with shugoshin[J ] . Tumor Biol, 2015, 36(8): 6067-6074. DOI: 10.1007/s13277-015-3284-7.

LI T W, FAN J Y, WANG B B, et al . TIMER: a web server for comprehensive analysis of tumor-infiltrating immune cells[J ] . Cancer Res, 2017, 77(21): e108-e110. DOI: 10.1158/0008-5472.CAN-17-0307.

TANG Z F, KANG B X, LI C W, et al . GEPIA2: an enhanced web server for large-scale expression profiling and interactive analysis[J ] . Nucleic Acids Res, 2019, 47(W1): W556-W560. DOI: 10.1093/nar/gkz430.

VASAIKAR S V, STRAUB P, WANG J, et al . LinkedOmics: analyzing multi-omics data within and across 32 cancer types[J ] . Nucleic Acids Res, 2018, 46(D1): D956-D963. DOI: 10.1093/nar/gkx1090.

ROUPRÊT M, BABJUK M, COMPÉRAT E, et al. European Association of Urology guidelines on upper urinary tract urothelial carcinoma: 2017 update[J ] . Eur Urol, 2018, 73(1): 111-122. DOI: 10.1016/j.eururo.2017.07.036.

BYUN S J, KIM J H, OH Y K, et al . Concurrent chemoradi otherapy improves survival outcome in muscle-invasive bladder cancer[J ] . Radiat Oncol J, 2015, 33(4): 294-300. DOI: 10.3857/roj.2015.33.4.294.

CHISM D D. Urothelial carcinoma of the bladder and the rise of immunotherapy[J]. J Natl Compr Canc Netw, 2017, 15(10): 1277-1284. DOI: 10.6004/jnccn.2017.7036.

SARI MOTLAGH R, MORI K, MIURA N, et al . The recurrence and progression risk after simultaneous endoscopic surgery of urothelial bladder tumour and benign prostatic hyperplasia: a systematic review and meta-analysis[J ] . BJU Int, 2021, 127(2): 143-152. DOI: 10.1111/bju.15146.

FERNÁNDEZ-CONEJO G, DE LA PEÑA E, HERNÁNDEZ V, et al . The value of tumour weight as a predictive factor for recurrence and progression in non-muscle invasive bladder cancer[J ] . Scand J Urol, 2020, 54(1): 40-45. DOI: 10.1080/21681805.2019.1708968.

TATEMATSU M, FUNAMI K, ISHII N, et al . LRRC59 regulates trafficking of nucleic acid-sensing TLRs from the endoplasmic reticulum via association with UNC93B1[J ] . J Immunol, 2015, 195(10): 4933-4942. DOI: 10.4049/jimmunol.1501305.

LI D, XING Y, TIAN T N, et al . Overexpression of LRRC59 is associated with poor prognosis and promotes cell proliferation and invasion in lung adenocarcinoma[J ] . Onco Targets Ther, 2020, 13: 6453-6463. DOI: 10.2147/OTT.S245336.

KIM J, KANG J, KANG Y L, et al . Ketohexokinase-a acts as a nuclear protein kinase that mediates fructose-induced metastasis in breast cancer[J ] . Nat Commun, 2020, 11(1): 5436. DOI: 10.1038/s41467-020-19263-1.

ZHEN Y, SØRENSEN V, SKJERPEN C S, et al . Nuclear import of exogenous FGF1 requires the ER-protein LRRC59 and the importins Kpnα1 and Kpnβ1[J ] . Traffic, 2012, 13 (5): 650-664. DOI: 10.1111/j.1600-0854.2012.01341.x.

TURNER N, GROSE R. Fibroblast growth factor signalling: from development to cancer[J]. Nat Rev Cancer, 2010, 10(2): 116-129. DOI: 10.1038/nrc2780.

PIROU C, MONTAZER-TORBATI F, JAH N, et al . FGF1 protects neuroblastoma SH-SY5Y cells from p53-dependent apoptosis through an intracrine pathway regulated by FGF1 phosphorylation[J ] . Cell Death Dis, 2017, 8(8): e3023. DOI: 10.1038/cddis.2017.404.

MAREI H E, ALTHANI A, AFIFI N, et al . p53 signaling in cancer progression and therapy[J ] . Cancer Cell Int, 2021, 21(1): 703. DOI: 10.1186/s12935-021-02396-8.

LEVINE A J. p53, the cellular gatekeeper for growth and division[J]. Cell, 1997, 88(3): 323-331. DOI: 10.1016/s0092-8674(00)81871-1.

SHI C L, SUN L, LIU S Z, et al . Overexpression of karyopherin subunit alpha 2 (KPNA2) predicts unfavorable prognosis and promotes bladder cancer tumorigenicity via the P53 pathway[J ] . Med Sci Monit, 2020, 26: e921087. DOI: 10.12659/MSM.921087.

ISLAM S S, MOKHTARI R B, EL HOUT Y, et al . TGF-β1 induces EMT reprogramming of porcine bladder urothelial cells into collagen producing fibroblasts-like cells in a Smad2/Smad3-dependent manner[J ] . J Cell Commun Signal, 2014, 8(1): 39-58. DOI: 10.1007/s12079-013-0216-4.

MCCONKEY D J, CHOI W, MARQUIS L, et al . Role of epithelial-to-mesenchymal transition (EMT) in drug sensitivity and metastasis in bladder cancer[J ] . Cancer Metastasis Rev, 2009, 28(3/4): 335-344. DOI: 10.1007/s10555-009-9194-7.

YU Q C, ZHANG K J, WANG X S, et al . Expression of transcript ion factors snail, slug, and twist in human bladder carcinoma[J ] . J Exp Clin Cancer Res, 2010, 29(1): 119. DOI: 10.1186/1756-9966-29-119.

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