GPRC5A调控的ABCB1表达对肺腺癌增殖的影响

李鋆; 崔雯雯; 杨中法; 刘文豪; 边茂旺; 邓炯; 王彤

doi:10.24920/004216

Your Location：

Home >

Browse articles >

GPRC5A调控的ABCB1表达对肺腺癌增殖的影响

中国医学科学杂志（英文） 2024年39卷第1期页码：9-18

论著 | Updated：2024-05-27

- GPRC5A调控的ABCB1表达对肺腺癌增殖的影响
- Affiliations：
  
  1. 1Department of Physiology, School of Basic Medical Sciences, Shandong Second Medical University, Weifang 261021, Shandong, China
  2. 2Medical Research Center, Affiliated Hospital of Binzhou Medical University, Binzhou 256600, Shandong, China
- Author bio：
  
  Jiong Deng, jiongdeng@bzmc.edu.cn.
  * E-mail: Tong Wang, wangtong@wfmc.edu.cn;
- Funds：
  
  国家自然科学基金(82103571);山东省自然科学基金青年项目(ZR2021QH295)
- DOI：10.24920/004216
  中图分类号：
- 收稿日期：2023-02-15，
  
  录用日期：2023-12-23，
  
  网络出版日期：2024-03-01，
  
  纸质出版日期：2024-03-31
- Accepted：
Scan QR Code
李鋆, 崔雯雯, 杨中法, 等. GPRC5A调控的ABCB1表达对肺腺癌增殖的影响[J]. 中国医学科学杂志（英文）, 2024,39(1):9-18.

Yun Li, Wen-Wen Cui, Zhong-Fa Yang, et al. Influence of GPRC5A-Regulated ABCB1 Expression on Lung Adenocarcinoma Proliferation[J]. Chinese medical sciences journal, 2024, 39(1): 9-18.
李鋆, 崔雯雯, 杨中法, 等. GPRC5A调控的ABCB1表达对肺腺癌增殖的影响[J]. 中国医学科学杂志（英文）, 2024,39(1):9-18. DOI： 10.24920/004216.

Yun Li, Wen-Wen Cui, Zhong-Fa Yang, et al. Influence of GPRC5A-Regulated ABCB1 Expression on Lung Adenocarcinoma Proliferation[J]. Chinese medical sciences journal, 2024, 39(1): 9-18. DOI： 10.24920/004216.

摘要

目的

ATP结合盒B亚家族成员 1（ATP binding cassette subfamily B member 1，ABCB1）的异常表达在多种癌症的发生发展中发挥关键作用。然而，G蛋白偶联受体C家族5组A型（G protein coupled receptor family C group 5 type A，GPRC5A）调控的ABCB1表达对肺腺癌增殖的影响仍不清楚。本研究探讨了GPRC5A调控的ABCB1表达对肺腺癌增殖的影响。

方法

我们采用RT-PCR、Western-blot或免疫组化实验，分析ABCB1在肺腺癌细胞系、人肺腺癌组织以及GPRC5A基因敲除小鼠和野生型小鼠的气管上皮细胞和肺组织中的表达。采用细胞计数试剂盒-8（CCK-8）分析GPRC5A基因敲除小鼠气管上皮细胞对化疗药物的敏感性。采用皮下肿瘤形成实验探讨下调ABCB1表达是否可抑制体内肺腺癌增殖。采用免疫荧光和免疫沉淀实验研究GPRC5A和ABCB1之间潜在的调控关系。

结果

ABCB1在肺腺癌细胞系和人类肺腺癌组织中表达上调。GPRC5A基因敲除小鼠的气管上皮细胞及肺组织的ABCB1表达高于野生型小鼠。与GPRC5A野生型小鼠的气管上皮细胞相比，GPRC5A基因敲除小鼠的气管上皮细胞对塔立奇达和多柔比星更敏感。注射移植细胞28天后，接受ABCB1基因敲除细胞移植的GPRC5A-/- C57BL/6小鼠的肺肿瘤的体积和重量均明显低于野生型细胞移植小鼠（

= 0.0043，

= 0.0060）。此外，免疫荧光和免疫沉淀实验表明，GPRC5A通过直接结合方式调控ABCB1的表达。

结论

GPRC5A通过抑制ABCB1表达降低肺腺癌增殖。GPRC5A调节ABCB1表达的途径有待研究。

Abstract

Objective

Aberrant expression of ATP binding cassette subfamily B member 1 (

ABCB1

) plays a key role in several cancers. However

influence of G protein coupled receptor family C group 5 type A (GPRC5A)-regulated

ABCB1

expression on lung adenocarcinoma proliferation remains unclea

r. Therefore

this study investigated the effect of

GPRC5A

regulated

ABCB1

expression on the proliferation of lung adenocarcinoma.

Methods

ABCB1

expressions in lung adenocarcinoma cell lines

human lung adenocarcinoma tissues

and tracheal epithelial cells and lung tissues of

GPRC5A

knockout mice and wild-type mice were analyzed with RT-PCR

Western blot

or immunohistochemical analysis. Cell counting kit-8 assay was performed to analyze the sensitivity of tracheal epithelial cells from

GPRC5A

knockout mice to chemotherapeutic agents. Subcutaneous tumor formation assay was performed to confirm whether down-regulation of

ABCB1

could inhibit the proliferation of lung adenocarcinoma

in vivo

. To verify the potential regulatory relationship between GPRC5A and ABCB1

immunofluorescence and immunoprecipitation assays were performed.

Results

ABCB1 expression was up-regulated in lung adenocarcinoma cell lines and human lung adenocarcinoma tissues. ABCB1 expression in the tracheal epithelial cells and lung tissues of

GPRC5A

deficient mice was higher than that in the wild type mice. Tracheal epithelial cells of

GPRC5A

knockout mice were much more sensitive to tariquidar and doxorubicin than those of

GPRC5A

wild type mice. Accordingly

28 days after injection of the transplanted cells

the volume and weight of lung tumor in

ABCB1

knockout cell-transplanted GPRC5A

-/-

C57BL/6 mice were significantly smaller than those in wild type cell-transplanted mice (

= 0.0043

= 0.0060). Furthermore

immunofluorescence and immunoprecipitation assays showed that GPRC5A regulated ABCB1 expression by direct binding.

Conclusion

GPRC5A reduces lung adenocarcinoma proliferation

via

inhibiting

ABCB1

expression. The pathway by which GPRC5A regulates

ABCB1

expression needs to be investigated.

关键词

Keywords

references

Zhang L , Li Y , Hu C , et al. CDK6-PI3K signaling axis is an efficient target for attenuating ABCB1/P-gp mediated multi-drug resistance (MDR) in cancer cells . Mol Cancer 2022 ; 21:103. doi: 10.1186/s12943-022-01524-w https://dx.doi.org/10.1186/s12943-022-01524-w .

Schinkel AH , Jonker JW . Mammalian drug efflux transporters of the ATP binding cassette (ABC) family: an overview . Adv Drug Deliv Rev 2003 ; 55 : 3 - 29 . doi: 10.1016/s0169-409x(02)00169-2 https://dx.doi.org/10.1016/s0169-409x(02)00169-2 . https://linkinghub.elsevier.com/retrieve/pii/S0169409X02001692 https://linkinghub.elsevier.com/retrieve/pii/S0169409X02001692

Katoh SY , Ueno M , Takakura N . Involvement of MDR1 function in proliferation of tumour cells . J Biochem 2008 ; 143 : 517 - 24 . doi: 10.1093/jb/mvm242 https://dx.doi.org/10.1093/jb/mvm242 . https://academic.oup.com/jb/article-lookup/doi/10.1093/jb/mvm242 https://academic.oup.com/jb/article-lookup/doi/10.1093/jb/mvm242

Smyth MJ , Krasovskis E , Sutton VR , et al. The drug efflux protein, P-glycoprotein, additionally protects drug-resistant tumor cells from multiple forms of caspase-dependent apoptosis . Proc Natl Acad Sci USA 1998 ; 95 : 7024 - 9 . doi: 10.1073/pnas.95.12.7024 https://dx.doi.org/10.1073/pnas.95.12.7024 .

Tao LY , Liang YJ , Wang F , et al. Cediranib (recentin, AZD2171) reverses ABCB1- and ABCC1-mediated multidrug resistance by inhibition of their transport function . Cancer Chemother Pharmacol 2009 ; 64:961-9. doi: 10.1007/s00280-009-0949-1 https://dx.doi.org/10.1007/s00280-009-0949-1 .

Eckford PD , Sharom FJ . ABC efflux pump-based resistance to chemotherapy drugs . Chem Rev 2009 ; 109 : 2989 - 3011 . doi: 10.1021/cr9000226 https://dx.doi.org/10.1021/cr9000226 .

Dong XD , Zhang M , Ma X , et al. Bruton’s tyrosine kinase (BTK) inhibitor RN486 overcomes ABCB1-mediated multidrug resistance in cancer cells . Front Cell Dev Biol 2020 ; 8:865. doi: 10.3389/fcell.2020.00865 https://dx.doi.org/10.3389/fcell.2020.00865 .

Zou T , Zeng C , Qu J , et al. Rutaecarpine increases anticancer drug sensitivity in drug-resistant cells through MARCH8-dependent ABCB1 degradation . Biomedicines 2021 ; 9(9):1143. doi: 10.3390/biomedicines9091143 https://dx.doi.org/10.3390/biomedicines9091143 .

Wu X , Yin C , Ma J , et al. Polyoxypregnanes as safe, potent, and specific ABCB1-inhibitory pro-drugs to overcome multidrug resistance in cancer chemotherapy in vitro and in vivo . Acta Pharm Sin B 2021 ; 11 ( 7 ): 1885 - 902 . doi: 10.1016/j.apsb.2020.12.021 https://dx.doi.org/10.1016/j.apsb.2020.12.021 . https://linkinghub.elsevier.com/retrieve/pii/S2211383521000034 https://linkinghub.elsevier.com/retrieve/pii/S2211383521000034

Zhou HH , Chen X , Cai LY , et al. Erastin reverses ABCB1-mediated docetaxel resistance in ovarian cancer . Front Oncol 2019 ; 9 : 1398 . doi: 10.3389/fonc.2019.01398 https://dx.doi.org/10.3389/fonc.2019.01398 . https://www.frontiersin.org/article/10.3389/fonc.2019.01398/full https://www.frontiersin.org/article/10.3389/fonc.2019.01398/full

Luo X , Teng QX , Dong JY , et al. Antimicrobial peptide reverses ABCB1-mediated chemotherapeutic drug resistance . Front Pharmacol 2020 ; 11 : 1208 . doi: 10.3389/fphar.2020.01208 https://dx.doi.org/10.3389/fphar.2020.01208 .

Cheng Y , Lotan R . Molecular cloning and characterization of a novel retinoic acid-inducible gene that encodes a putative G protein-coupled receptor . JBC 1998 ; 273 ( 52 ): 35008 - 15 . doi: 10.1074/jbc.273.52.35008 https://dx.doi.org/10.1074/jbc.273.52.35008 . https://linkinghub.elsevier.com/retrieve/pii/S0021925818371758 https://linkinghub.elsevier.com/retrieve/pii/S0021925818371758

Tao Q , Fujimoto J , Men T , et al. Identification of the retinoic acid-inducible Gprc5a as a new lung tumor suppressor gene . J Natl Cancer Inst 2007 ; 99 ( 22 ): 1668 - 82 . doi: 10.1093/jnci/djm208 https://dx.doi.org/10.1093/jnci/djm208 .

Xu J , Tian J , Shapiro SD . Normal lung development in RAIG1-deficient mice despite unique lung epithelium-specific expression . Am J Respir Cell Mol Biol 2005 ; 32 ( 5 ): 381 - 7 . doi: 10.1165/rcmb.2004-0343OC https://dx.doi.org/10.1165/rcmb.2004-0343OC . https://www.atsjournals.org/doi/10.1165/rcmb.2004-0343OC https://www.atsjournals.org/doi/10.1165/rcmb.2004-0343OC

Song H , Ye X , Liao Y , et al. NF-κB represses retinoic acid receptor-mediated GPRC5A transactivation in lung epithelial cells to promote neoplasia . JCI Insight 2023 ; 8(1):e153976. doi: 10.1172/jci.insight.153976 https://dx.doi.org/10.1172/jci.insight.153976 .

Zhong S , Yin H , Liao Y , et al. Lung tumor suppressor GPRC5A binds EGFR and restrains its effector signaling . Cancer Res 2015 ; 75 ( 9 ): 1801 - 14 . doi: 10.1158/0008-5472.CAN-14-2005 https://dx.doi.org/10.1158/0008-5472.CAN-14-2005 .

Wang T , Jing B , Xu D , et al. PTGES/PGE(2) signaling links immunosuppression and lung metastasis in Gprc5a-knockout mouse model . Oncogene 2020 ; 39(15):3179-94. doi: 10.1038/s41388-020-1207-6 https://dx.doi.org/10.1038/s41388-020-1207-6 .

Olarte Carrillo I , García Laguna AI , De la Cruz Rosas A , et al. High expression levels and the C3435T SNP of the ABCB 1 gene are associated with lower survival in adult patients with acute myeloblastic leukemia in Mexico City . BMC Med Genomics 2021 ; 14(1):251. doi: 10.1186/s12920-021-01101-y https://dx.doi.org/10.1186/s12920-021-01101-y .

Jadhao M , Tsai EM , Yang HC , et al. The long-term DEHP exposure confers multidrug resistance of triple-negative breast cancer cells through ABC transporters and intracellular ROS . Antioxidants (Basel, Switzerland) 2021 ; 10 ( 6 ): 102021 . doi: 10.3390/antiox10060949 https://dx.doi.org/10.3390/antiox10060949 .

Goto S , Kawabata T , Li TS . Enhanced expression of ABCB1 and Nrf2 in CD133-positive cancer stem cells associates with doxorubicin resistance . Stem Cells Int 2020 ; 2020:8868849. doi: 10.1155/2020/8868849 https://dx.doi.org/10.1155/2020/8868849 .

Nasir A , Cardall A , Othman RT , et al. ABCB1 inhibition provides a novel therapeutic target to block TWIST1-induced migration in medulloblastoma . Neurooncol Adv 2021 ; 3(1):vdab030. doi: 10.1093/noajnl/vdab030 https://dx.doi.org/10.1093/noajnl/vdab030 .

Liu D , Jin Y , Wu J , et al. MiR-135b-5p is an oncogene in pancreatic cancer to regulate GPRC5A expression by targeting transcription factor KLF4 . Cell Death Discov 2022 ; 8(1):23. doi: 10.1038/s41420-022-00814-y https://dx.doi.org/10.1038/s41420-022-00814-y .

Sokolenko AP , Bulanova DR , Iyevleva AG , et al. High prevalence of GPRC5A germline mutations in BRCA1-mutant breast cancer patients . Int J Cancer 2014 ; 134(10):2352-8.. doi: 10.1002/ijc.28569 https://dx.doi.org/10.1002/ijc.28569 .

Ufer M , Hasler R , Jacobs G , et al. Decreased sigmoidal ABCB 1 (P-glycoprotein) expression in ulcerative colitis is associated with disease activity . Pharmacogenomics 2009 ; 10 ( 12 ): 1941 - 53 . doi: 10.2217/pgs.09.128 https://dx.doi.org/10.2217/pgs.09.128 . https://www.futuremedicine.com/doi/10.2217/pgs.09.128 https://www.futuremedicine.com/doi/10.2217/pgs.09.128

Nagahata T , Sato T , Tomura A , et al. Identification of RAI3 as a therapeutic target for breast cancer . Endocr Relat Cancer 2005 ; 12 ( 1 ): 65 - 73 . doi: 10.1677/erc.1.00890 https://dx.doi.org/10.1677/erc.1.00890 . https://erc.bioscientifica.com/doi/10.1677/erc.1.00890 https://erc.bioscientifica.com/doi/10.1677/erc.1.00890

Zhou H , Rigoutsos I . The emerging roles of GPRC5A in diseases . Oncoscience 2012 ; 1 ( 12 ): 765 - 76 . doi: 10.18632/oncoscience.104 https://dx.doi.org/10.18632/oncoscience.104 . https://www.oncoscience.us/lookup/doi/10.18632/oncoscience.v1i12 https://www.oncoscience.us/lookup/doi/10.18632/oncoscience.v1i12

Yang L , Zhao S , Zhu T , et al. GPRC5A is a negative regulator of the pro-survival PI3K/Akt signaling pathway in triple-negative breast cancer . Front Oncol 2020 ; 10:624493. doi: 10.3389/fonc.2020.624493 https://dx.doi.org/10.3389/fonc.2020.624493 .

浏览量

865

Downloads

CSCD

文章被引用时，请邮件提醒。

Submit

关联资源

Prognostic Prediction Value and Biological Functions of Non-Apoptotic Regulated Cell Death Genes in Lung Adenocarcinoma

Identifying and Validating a Novel miRNA-mRNA Regulatory Network Associated with Prognosis in Lung Adenocarcinoma