Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer

Feng,Yu-Zhen; Zhang,Qing-Yan; Fu,Mei-Ting; Zhang,Zhen-Fei; Wei,Min; Zhou,Jue-Yu; Shi,Rong

doi:10.3892/or.2017.5696

Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer

Authors:
- Yu-Zhen Feng
- Qing-Yan Zhang
- Mei-Ting Fu
- Zhen-Fei Zhang
- Min Wei
- Jue-Yu Zhou
- Rong Shi
View Affiliations

Affiliations: Institute of Genetic Engineering, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, The First Clinical Medical College, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China
Published online on: June 2, 2017 https://doi.org/10.3892/or.2017.5696
Pages: 109-119
Copyright: © Feng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Human Pinx1 protein, associated with shelterin proteins, is widely revealed as a haploinsufficient tumor suppressor. Growing evidence has manifested the deregulation of PinX1 in distinct cancers. Nonetheless, the loss status of PinX1 and its diagnostic, prognostic and clinicopathological significance in Basal-like breast cancer are still unclear. In the present study, the PinX1 expression levels of breast cancer tissues were investigated by qRT-PCR and immunoblotting assays. Then immunohistochemistry (IHC) was performed to detect PinX1 expression on a tissue microarray. The optimal threshold for PinX1 positivity was determined by receiver operating characteristic (ROC) curve analysis. To clarify the probable role of PinX1 in BLBC, the PinX1 knockout and stably over-expressed MDA-MB-231 cell lines were constructed by the CRISPR-Cas9 system and gene transfection. The association of PinX1 expression with cell proliferation, migration and apoptosis of MDA-MB-231 cells were observed by CCK-8 assay, wound healing assay, transwell assay, flow cytometric analysis and immunoblotting of the cleaved caspase-3 protein level. Our results showed that both PinX1 mRNA and protein expression were downregulated in breast cancer tissues (P<0.05). In IHC analysis, the optimal cut-off parameter for PinX1 positive expression was 62.5% (the AUC was 0.749, P<0.01). PinX1 positivity was 76.9% (10/14) in luminal subtypes, 50% (5/10) in Her2-enriched breast cancer and 27.3% (9/33) in basal-like subtypes. Besides, in 59 invasive ductal breast carcinomas, PinX1 expression was inversely related to histology grade (P<0.05) while it was positively associated with PR status (P<0.05) and ER status (P<0.05). These results indicated that low expression of PinX1 correlated with aggressive clinicopathological significance of breast cancer, especially in the basal-like subtype. Besides, we identified that overexpression of PinX1 inhibited the proliferation rates and migration ability and increased the apoptosis rates of BLBC. Our findings demonstrated that low expression of PinX1 was associated with malignant behaviors in basal-like subtype of breast cancer. PinX1 is likely a feasible biomarker and molecular target of BLBC.

View Figures

View References

1	Taherian-Fard A, Srihari S and Ragan MA: Breast cancer classification: linking molecular mechanisms to disease prognosis. Brief Bioinform. 16:461–474. 2015. View Article : Google Scholar : PubMed/NCBI
2	Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D and Bray F: Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 136:E359–E386. 2015. View Article : Google Scholar : PubMed/NCBI
3	Goldhirsch A, Winer EP, Coates AS, Gelber RD, Piccart-Gebhart M, Thürlimann B, Senn HJ, Albain KS, André F, Bergh J, et al: Panel members: Personalizing the treatment of women with early breast cancer: Highlights of the St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2013. Ann Oncol. 24:2206–2223. 2013. View Article : Google Scholar : PubMed/NCBI
4	Vallejos CS, Gómez HL, Cruz WR, Pinto JA, Dyer RR, Velarde R, Suazo JF, Neciosup SP, León M, de la Cruz MA, et al: Breast cancer classification according to immunohistochemistry markers: Subtypes and association with clinicopathologic variables in a Peruvian hospital database. Clin Breast Cancer. 10:294–300. 2010. View Article : Google Scholar : PubMed/NCBI
5	Ramirez-Fort MK, Case EC, Rosen AC, Cerci FB, Wu S and Lacouture ME: Rash to the mTOR inhibitor everolimus: Systematic review and meta-analysis. Am J Clin Oncol. 37:266–271. 2014. View Article : Google Scholar : PubMed/NCBI
6	Gelmon K, Dent R, Mackey JR, Laing K, McLeod D and Verma S: Targeting triple-negative breast cancer: Optimising therapeutic outcomes. Ann Oncol. 23:2223–2234. 2012. View Article : Google Scholar : PubMed/NCBI
7	Bianchini G, Balko JM, Mayer IA, Sanders ME and Gianni L: Triple-negative breast cancer: Challenges and opportunities of a heterogeneous disease. Nat Rev Clin Oncol. 13:674–690. 2016. View Article : Google Scholar : PubMed/NCBI
8	Perou CM: Molecular stratification of triple-negative breast cancers. Oncologist. 15 Suppl 5:39–48. 2010. View Article : Google Scholar : PubMed/NCBI
9	De Laurentiis M, Cianniello D, Caputo R, Stanzione B, Arpino G, Cinieri S, Lorusso V and De Placido S: Treatment of triple negative breast cancer (TNBC): Current options and future perspectives. Cancer Treat Rev. 36 Suppl 3:S80–S86. 2010. View Article : Google Scholar : PubMed/NCBI
10	Eroles P, Bosch A, Pérez-Fidalgo JA and Lluch A: Molecular biology in breast cancer: Intrinsic subtypes and signaling pathways. Cancer Treat Rev. 38:698–707. 2012. View Article : Google Scholar : PubMed/NCBI
11	Esposito A: Highlights from the 14th St Gallen International Breast Cancer Conference 2015 in Vienna: Dealing with classification, prognostication, and prediction refinement to personalize the treatment of patients with early breast cancer. E Cancer Med Sci. 9:5182015.
12	Nelson ER, Li S, Kennedy M, Payne S, Kilibarda K, Groth J, Bowie M, Parilla-Castellar E, De Ridder G, Marcom PK, et al: Chemotherapy enriches for an invasive triple-negative breast tumor cell subpopulation expressing a precursor form of N-cadherin on the cell surface. Oncotarget. 7:84030–84042. 2016.PubMed/NCBI
13	Rodler ET, Kurland BF, Griffin M, Gralow JR, Porter P, Yeh RF, Gadi VK, Guenthoer J, Beumer JH, Korde L, et al: Phase I study of veliparib (ABT-888) combined with cisplatin and vinorelbine in advanced triple-negative breast cancer and/or BRCA mutation-associated breast cancer. Clin Cancer Res. 22:2855–2864. 2016. View Article : Google Scholar : PubMed/NCBI
14	Li Y and Tergaonkar V: Noncanonical functions of telomerase: Implications in telomerase-targeted cancer therapies. Cancer Res. 74:1639–1644. 2014. View Article : Google Scholar : PubMed/NCBI
15	Zhou XZ, Huang P, Shi R, Lee TH, Lu G, Zhang Z, Bronson R and Lu KP: The telomerase inhibitor PinX1 is a major haploinsufficient tumor suppressor essential for chromosome stability in mice. J Clin Invest. 121:1266–1282. 2011. View Article : Google Scholar : PubMed/NCBI
16	Tian XP, Qian D, He LR, Huang H, Mai SJ, Li CP, Huang XX, Cai MY, Liao YJ, Kung HF, et al: The telomere/telomerase binding factor PinX1 regulates paclitaxel sensitivity depending on spindle assembly checkpoint in human cervical squamous cell carcinomas. Cancer Lett. 353:104–114. 2014. View Article : Google Scholar : PubMed/NCBI
17	Liu JY, Qian D, He LR, Li YH, Liao YJ, Mai SJ, Tian XP, Liu YH, Zhang JX, Kung HF, et al: PinX1 suppresses bladder urothelial carcinoma cell proliferation via the inhibition of telomerase activity and p16/cyclin D1 pathway. Mol Cancer. 12:1482013. View Article : Google Scholar : PubMed/NCBI
18	Li HL, Han L, Chen HR, Meng F, Liu QH, Pan ZQ, Bai J and Zheng JN: PinX1 serves as a potential prognostic indicator for clear cell renal cell carcinoma and inhibits its invasion and metastasis by suppressing MMP-2 via NF-κB-dependent transcription. Oncotarget. 6:21406–21420. 2015. View Article : Google Scholar : PubMed/NCBI
19	Shi R, Zhao Z, Zhou H, Wei M, Ma WL, Zhou JY and Tan WL: Reduced expression of PinX1 correlates to progressive features in patients with prostate cancer. Cancer Cell Int. 14:462014. View Article : Google Scholar : PubMed/NCBI
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2ΔΔCT method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
21	Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA and Zhang F: Genome engineering using the CRISPR-Cas9 system. Nat Protoc. 8:2281–2308. 2013. View Article : Google Scholar : PubMed/NCBI
22	Lai XF, Shen CX, Wen Z, Qian YH, Yu CS, Wang JQ, Zhong PN and Wang HL: PinX1 regulation of telomerase activity and apoptosis in nasopharyngeal carcinoma cells. J Exp Clin Cancer Res. 31:122012. View Article : Google Scholar : PubMed/NCBI
23	Toss A and Cristofanilli M: Molecular characterization and targeted therapeutic approaches in breast cancer. Breast Cancer Res. 17:602015. View Article : Google Scholar : PubMed/NCBI
24	Morris PG, Murphy CG, Mallam D, Accordino M, Patil S, Howard J, Omuro A, Beal K, Seidman AD, Hudis CA, et al: Limited overall survival in patients with brain metastases from triple negative breast cancer. Breast J. 18:345–350. 2012. View Article : Google Scholar : PubMed/NCBI
25	Ogata H, Kikuchi Y, Natori K, Shiraga N, Kobayashi M, Magoshi S, Saito F, Osaku T, Kanazawa S, Kubota Y, et al: Liver metastasis of a triple-negative breast cancer and complete remission for 5 years after treatment with combined bevacizumab/paclitaxel/carboplatin: Case Report and Review of the Literature. Medicine (Baltimore). 94:e17562015. View Article : Google Scholar : PubMed/NCBI
26	Bartholomeusz C, Xie X, Pitner MK, Kondo K, Dadbin A, Lee J, Saso H, Smith PD, Dalby KN and Ueno NT: MEK inhibitor selumetinib (AZD6244; ARRY-142886) prevents lung metastasis in a triple-negative breast xancer xenograft model. Mol Cancer Ther. 14:2773–2781. 2015. View Article : Google Scholar : PubMed/NCBI
27	Burnett JP, Korkaya H, Ouzounova MD, Jiang H, Conley SJ, Newman BW, Sun L, Connarn JN, Chen CS, Zhang N, et al: Trastuzumab resistance induces EMT to transform HER2+ PTEN to a triple negative breast cancer that requires unique treatment options. Sci Rep. 5:158212015. View Article : Google Scholar : PubMed/NCBI
28	Pal SK, Childs BH and Pegram M: Triple negative breast cancer: Unmet medical needs. Breast Cancer Res Treat. 125:627–636. 2011. View Article : Google Scholar : PubMed/NCBI
29	Wu J, Li S, Jia W and Su F: Response and prognosis of taxanes and anthracyclines neoadjuvant chemotherapy in patients with triple-negative breast cancer. J Cancer Res Clin Oncol. 137:1505–1510. 2011. View Article : Google Scholar : PubMed/NCBI
30	Carey LA, Rugo HS, Marcom PK, Mayer EL, Esteva FJ, Ma CX, Liu MC, Storniolo AM, Rimawi MF, Forero-Torres A, et al: TBCRC 001: Randomized phase II study of cetuximab in combination with carboplatin in stage IV triple-negative breast cancer. J Clin Oncol. 30:2615–2623. 2012. View Article : Google Scholar : PubMed/NCBI
31	Lang JE, Wecsler JS, Press MF and Tripathy D: Molecular markers for breast cancer diagnosis, prognosis and targeted therapy. J Surg Oncol. 111:81–90. 2015. View Article : Google Scholar : PubMed/NCBI
32	Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PL, Coviello GM, Wright WE, Weinrich SL and Shay JW: Specific association of human telomerase activity with immortal cells and cancer. Science. 266:2011–2015. 1994. View Article : Google Scholar : PubMed/NCBI
33	Johnson FB: PinX1 the tail on the chromosome. J Clin Invest. 121:1242–1244. 2011. View Article : Google Scholar : PubMed/NCBI
34	Artandi SE and DePinho RA: Telomeres and telomerase in cancer. Carcinogenesis. 31:9–18. 2010. View Article : Google Scholar : PubMed/NCBI
35	Deng W, Jiao N, Li N, Wan X, Luo S and Zhang Y: Decreased expression of PinX1 protein predicts poor prognosis of colorectal cancer patients receiving 5-FU adjuvant chemotherapy. Biomed Pharmacother. 73:1–5. 2015. View Article : Google Scholar : PubMed/NCBI
36	Qian D, Zhang B, He LR, Cai MY, Mai SJ, Liao YJ, Liu YH, Lin MC, Bian XW, Zeng YX, et al: The telomere/telomerase binding factor PinX1 is a new target to improve the radiotherapy effect of oesophageal squamous cell carcinomas. J Pathol. 229:765–774. 2013. View Article : Google Scholar : PubMed/NCBI
37	Wang HB, Wang XW, Zhou G, Wang WQ, Sun YG, Yang SM and Fang DC: PinX1 inhibits telomerase activity in gastric cancer cells through Mad1/c-Myc pathway. J Gastrointest Surg. 14:1227–1234. 2010. View Article : Google Scholar : PubMed/NCBI
38	Swanson PE: Immunohistochemistry as a surrogate for molecular testing: A review. Appl Immunohistochem Mol Morphol. 23:81–96. 2015. View Article : Google Scholar : PubMed/NCBI
39	Zuo J, Wang DH, Zhang YJ, Liu L, Liu FL and Liu W: Expression and mechanism of PinX1 and telomerase activity in the carcinogenesis of esophageal epithelial cells. Oncol Rep. 30:1823–1831. 2013.PubMed/NCBI