Genetic Ablation of Pregnancy Zone Protein Promotes Breast Cancer Progression by Activating TGF-β/SMAD Signaling

Overview

Journal Breast Cancer Res Treat

Specialty Oncology

Date 2020 Oct 15

PMID 33057846

Citations 3

Authors

Rohit Kumar

Ekaterina Kuligina

Anna Sokolenko

Quadir Siddiqui

Nilesh Gardi

Sudeep Gupta

Ashok K Varma

Syed K Hasan

Affiliations

Soon will be listed here.

Abstract

Purpose: Pregnancy zone protein (PZP) is best known as protease inhibitor and its concentration in human blood plasma increases dramatically during pregnancy. Recent investigation revealed a role of PZP inactivating germ-line mutation in breast cancer predisposition, and therefore we designed a study to evaluate functional involvement of this protein in tumor pathogenesis.

Methods: PZP knockout cells were generated utilizing the CRISPR-Cas9 approach in MCF7 and T47D (breast cancer) cell lines, and colony formation, cell proliferation, and migration assays carried out. TGF-β and SMAD expression studies were performed using qRT-PCR and Western blot. PZP expression in tumor vs normal tissue was compared using meta-analyses of data records of breast cancer patients (n = 1211) included in the TCGA consortium registry as well as in independent cohorts of hormone receptor-positive (n = 118) and triple-negative breast cancer (TNBC) patients (n = 116).

Results: We demonstrated that genetic ablation of PZP efficiently inhibits tamoxifen-induced apoptosis and enhances cell proliferation, migration, and colony-forming capacity. We found a significant increase in survival fraction of CRISPR/Cas9-mediated PZP knockout clones compared to wild-type counterpart after tamoxifen treatment (p < 0.05). The PZP knockout significantly promoted breast cancer cell migration (p < 0.01) in vitro. We observed high expression of TGF-β2 ligand, TGF-β- receptor 2, and upregulation of phosphorylated regulatory-SMADs (pSMAD2 and pSMAD3) activating the pro-survival function of TGF-β/SMAD signaling in PZP knockout clones. Meta-analyses of data records of breast cancer patients indicated that low PZP expression is associated with poor overall survival at 6 years (51.7% vs 62.9% in low vs high expressers, respectively; p = 0.026). We also observed a significantly lower PZP mRNA expression in TNBC as compared with hormone receptor-positive tumors (p = 0.019).

Conclusion: Taken together, our results suggest that genetic ablation of PZP results in tumor progression and low expression of PZP is associated with poor survival of breast cancer patients.

Citing Articles

Host-defence caerin 1.1 and 1.9 peptides suppress glioblastoma U87 and U118 cell proliferation through the modulation of mitochondrial respiration and induce the downregulation of CHI3L1.

Wang Y, Zhong F, Xiao F, Li J, Liu X, Ni G PLoS One. 2024; 19(6):e0304149.

PMID: 38848430 PMC: 11161062. DOI: 10.1371/journal.pone.0304149.

Pregnancy Zone Protein Serves as a Prognostic Marker and Favors Immune Infiltration in Lung Adenocarcinoma.

Chen K, Zheng T, Chen C, Liu L, Guo Z, Peng Y Biomedicines. 2023; 11(7).

PMID: 37509617 PMC: 10377424. DOI: 10.3390/biomedicines11071978.

Hepatokine Pregnancy Zone Protein Governs the Diet-Induced Thermogenesis Through Activating Brown Adipose Tissue.

Lin J, Jiang X, Dong M, Liu X, Shen Q, Huang Y Adv Sci (Weinh). 2021; 8(21):e2101991.

PMID: 34514733 PMC: 8564441. DOI: 10.1002/advs.202101991.

References

Siegel R, Miller K, Jemal A . Cancer statistics, 2019. CA Cancer J Clin. 2019; 69(1):7-34. DOI: 10.3322/caac.21551. View

Siegel R, Naishadham D, Jemal A . Cancer statistics, 2013. CA Cancer J Clin. 2013; 63(1):11-30. DOI: 10.3322/caac.21166. View

Torrezan G, de Almeida F, Figueiredo M, Barros B, de Paula C, Valieris R . Complex Landscape of Germline Variants in Brazilian Patients With Hereditary and Early Onset Breast Cancer. Front Genet. 2018; 9:161. PMC: 5949367. DOI: 10.3389/fgene.2018.00161. View

Kuligina E, Sokolenko A, Bizin I, Romanko A, Zagorodnev K, Anisimova M . Exome sequencing study of Russian breast cancer patients suggests a predisposing role for USP39. Breast Cancer Res Treat. 2019; 179(3):731-742. DOI: 10.1007/s10549-019-05492-6. View

Thompson E, Doyle M, Ryland G, Rowley S, Choong D, Tothill R . Exome sequencing identifies rare deleterious mutations in DNA repair genes FANCC and BLM as potential breast cancer susceptibility alleles. PLoS Genet. 2012; 8(9):e1002894. PMC: 3459953. DOI: 10.1371/journal.pgen.1002894. View

Wong S, Dessen A . Structure of a bacterial α2-macroglobulin reveals mimicry of eukaryotic innate immunity. Nat Commun. 2014; 5:4917. DOI: 10.1038/ncomms5917. View

Philip A, Bostedt L, Stigbrand T, OConnor-McCourt M . Binding of transforming growth factor-beta (TGF-beta) to pregnancy zone protein (PZP). Comparison to the TGF-beta-alpha 2-macroglobulin interaction. Eur J Biochem. 1994; 221(2):687-93. DOI: 10.1111/j.1432-1033.1994.tb18781.x. View

Zheng Y, Liu Y, Zhao S, Zheng Z, Shen C, An L . Large-scale analysis reveals a novel risk score to predict overall survival in hepatocellular carcinoma. Cancer Manag Res. 2018; 10:6079-6096. PMC: 6252784. DOI: 10.2147/CMAR.S181396. View

Zhang Z, Li J, He T, Ouyang Y, Huang Y, Liu Q . Two predictive precision medicine tools for hepatocellular carcinoma. Cancer Cell Int. 2019; 19:290. PMC: 6854692. DOI: 10.1186/s12935-019-1002-z. View

10.

Daroqui M, Vazquez P, Bal De Kier Joffe E, Bakin A, Puricelli L . TGF-β autocrine pathway and MAPK signaling promote cell invasiveness and in vivo mammary adenocarcinoma tumor progression. Oncol Rep. 2012; 28(2):567-75. PMC: 3981025. DOI: 10.3892/or.2012.1813. View

11.

Ran F, Hsu P, Wright J, Agarwala V, Scott D, Zhang F . Genome engineering using the CRISPR-Cas9 system. Nat Protoc. 2013; 8(11):2281-2308. PMC: 3969860. DOI: 10.1038/nprot.2013.143. View

12.

Mohr S, Hu Y, Ewen-Campen B, Housden B, Viswanatha R, Perrimon N . CRISPR guide RNA design for research applications. FEBS J. 2016; 283(17):3232-8. PMC: 5014588. DOI: 10.1111/febs.13777. View

13.

Dehairs J, Talebi A, Cherifi Y, Swinnen J . CRISP-ID: decoding CRISPR mediated indels by Sanger sequencing. Sci Rep. 2016; 6:28973. PMC: 4929496. DOI: 10.1038/srep28973. View

14.

Chen Y, Ma L, He Q, Zhang S, Zhang C, Jia W . TGF-β1 expression is associated with invasion and metastasis of intrahepatic cholangiocarcinoma. Biol Res. 2015; 48:26. PMC: 4513632. DOI: 10.1186/s40659-015-0016-9. View

15.

Ye J, Coulouris G, Zaretskaya I, Cutcutache I, Rozen S, Madden T . Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics. 2012; 13:134. PMC: 3412702. DOI: 10.1186/1471-2105-13-134. View

16.

Franken N, Rodermond H, Stap J, Haveman J, van Bree C . Clonogenic assay of cells in vitro. Nat Protoc. 2007; 1(5):2315-9. DOI: 10.1038/nprot.2006.339. View

17.

Rueden C, Schindelin J, Hiner M, DeZonia B, Walter A, Arena E . ImageJ2: ImageJ for the next generation of scientific image data. BMC Bioinformatics. 2017; 18(1):529. PMC: 5708080. DOI: 10.1186/s12859-017-1934-z. View

18.

Colak S, Ten Dijke P . Targeting TGF-β Signaling in Cancer. Trends Cancer. 2017; 3(1):56-71. DOI: 10.1016/j.trecan.2016.11.008. View

19.

Liu N, Xi Y, Callaghan M, Fribley A, Moore-Smith L, Zimmerman J . SMAD4 is a potential prognostic marker in human breast carcinomas. Tumour Biol. 2013; 35(1):641-50. PMC: 4142205. DOI: 10.1007/s13277-013-1088-1. View

20.

Stuelten C, Buck M, Dippon J, Roberts A, Fritz P, Knabbe C . Smad4-expression is decreased in breast cancer tissues: a retrospective study. BMC Cancer. 2006; 6:25. PMC: 1413545. DOI: 10.1186/1471-2407-6-25. View