Expression profiles of LMAN2 in breast cancer cell lines in hypoxia and normoxia conditions

Abstract

Abstract Purpose: Breast cancer is one of the most common and leading causes of death in women. The mechanism by which breast cancer develops is not fully understood. Understanding the mechanism of initiation and the genes and proteins involved in this process may help us to fight this type of cancer. The LMAN2 gene encodes VIP36, which transports properly folded proteins. The main aim of this study was to investigate the expression of the LMAN2 gene at the molecular level in breast cancer cells with different functional defects. Determining the level of LMAN2 gene expression under hypoxia, known as oxygen deprivation, has a significant impact on tumourigenesis and metastasis, and obtaining new data on the relationship between hypoxia and ER stress was identified as a secondary objective. Material and methods: In this study, the expression level of the LMAN2 gene will be examined in breast cancer cell lines (SKBR3, MDA-MB-231, MDA-MB-468, MCF-7) and CRL4010 cell line as a control. Results: LMAN2 gene expression level was evaluated at 48-hour periods by providing normoxic and hypoxic conditions. While the LMAN2 gene expression level in MCF-7, MDA-MB-231, and SK-BR-3 cells is significantly reduced, it was highly expressed in MDA-MB-468 in hypoxic and normoxic conditions. CHOP, HERP, and BiP gene expression levels were significantly higher in MDA-MB-468 under hypoxic conditions like LMAN2 expression. Conclusion: LMAN2 and other ER stress response elements showed different expression profiles in SK-BR-3, MDA-MB-231, MDA-MB-468, and MCF7 cell lines under hypoxic conditions. Increased expression was found in MCF-7 and MDA-MB-468 cell lines, but decreased expression was detected in SK-BR-3 and MDA-MB-231 cell lines. The underlying reason for this difference is thought to be that the cell lines have different molecular properties, such as triple negative or HER2 (+/-) status.

Keywords

• ER stress
• hypoxia
• LMAN2
• breast cancer
• apoptosis

Hipoksi ve normoksi koşullarında meme kanseri hücre hatlarında LMAN2'nin ifade profilleri

Abstract

Amaç: Meme kanseri kadınlarda en sık görülen ve ölüme neden olan kanser türleri arasındadır. Meme kanserinin başlama mekanizması tam olarak açık değildir. Ortaya çıkış mekanizmasını ve bu süreçte yer alan genleri ve proteinleri anlamak, bu kanser türüyle mücadelemizde yardımcı olabileceğini düşünmekteyiz. LMAN2 geni, doğru katlanmış proteinleri taşıyan VIP36'yı kodlar. Bu çalışmanın temel amacı, çeşitli fonksiyonel bozukluklara sahip meme kanseri hücrelerinde LMAN2 geninin ekspresyonunu moleküler düzeyde incelemektir. Oksijen eksikliği koşulları olarak bilinen hipoksi altında LMAN2 gen ekspresyon düzeyinin saptanması, tümörigenez ve metastaz üzerinde önemli bir etkiye sahiptir ve hipoksi ile ER stresi arasındaki ilişki hakkında yeni veriler elde edilmesi de ikincil bir hedef olarak belirlenmiştir. Gereç ve yöntem: Bu çalışmada, meme kanseri hücre hatlarında (SKBR3, MDA-MB-231, MDA-MB-468, MCF-7) ve kontrol olarak CRL4010 hücre hattında LMAN2 geninin ifade düzeyi incelendi. Bulgular: LMAN2 gen ekspresyon düzeyi normoksik ve hipoksik koşullar sağlanarak 48 saatlik periyotta değerlendirilmiştir. MCF-7, MDA-MB-231 ve SK-BR-3 hücrelerinde LMAN2 gen ekspresyon düzeyi önemli ölçüde azalırken, MDA-MB-468'de hipoksik ve normoksik koşullarda yüksek düzeyde eksprese edildi. CHOP, HERP ve BiP gen ekspresyon seviyeleri, LMAN2 ekspresyonu gibi hipoksik koşullar altında MDA-MB-468'de önemli ölçüde daha yüksekti. Sonuç: LMAN2 ve diğer ER stres yanıt elemanları hipoksik koşullar altında SK-BR-3, MDA-MB-231, MDA-MB-468 ve MCF7 hücre hatlarında farklı ekspresyon profilleri göstermiştir. MCF-7 ve MDA-MB-468 hücre hatlarında artmış ekspresyon bulunurken, SK-BR-3 ve MDA-MB-231 hücre hatlarında azalmış ekspresyon tespit edilmiştir. Bu farklılığın altında yatan nedenin, hücre hatlarının üçlü negatif veya HER2 (+/-) durumu gibi farklı moleküler özelliklere sahip olması olduğu düşünülmektedir.

Keywords

• ER stres
• hipoksi
• LMAN2
• meme kanseri
• apoptoz

References

1. 1. Banach A, Jiang YP, Roth E, Kuscu C, Cao J, Lin RZ. CEMIP upregulates BiP to promote breast cancer cell survival in hypoxia. Oncotarget. 2019;10(42):4307-4320. Published 2019 Jul 2. doi:10.18632/oncotarget.27036
2. 2. Siegel RL, Miller KD, Wagle NS, Jemal A. Cancer statistics, 2023. CA Cancer J Clin. 2023;73(1):17-48. doi:10.3322/caac.21763
3. 3. Eliyatkın N, Yalçın E, Zengel B, Aktaş S, Vardar E. Molecular Classification of Breast Carcinoma: From Traditional, Old-Fashioned Way to A New Age, and A New Way. J Breast Health. 2015;11(2):59-66. Published 2015 Apr 1. doi:10.5152/tjbh.2015.1669
4. 4. Landskron G, De la Fuente M, Thuwajit P, Thuwajit C, Hermoso MA. Chronic inflammation and cytokines in the tumor microenvironment. J Immunol Res. 2014;2014:149185. doi:10.1155/2014/149185
5. 5. Bettencourt C, Foti SC, Miki Y, et al. White matter DNA methylation profiling reveals deregulation of HIP1, LMAN2, MOBP, and other loci in multiple system atrophy. Acta Neuropathol. 2020;139(1):135-156. doi:10.1007/s00401-019-02074-0
6. 6. Davalieva K, Kiprijanovska S, Maleva Kostovska I, et al. Comparative Proteomics Analysis of Urine Reveals Down-Regulation of Acute Phase Response Signaling and LXR/RXR Activation Pathways in Prostate Cancer. Proteomes. 2017;6(1):1. Published 2017 Dec 29. doi:10.3390/proteomes6010001
7. 7. Demiryürek AN, Göktürk Ö, Saracaloglu A, Demiryürek S, Demiryürek AT. Protective effects of verbenalin and (+)-eudesmin against 6-hydroxydopamine-induced oxidative/nitrosative stress in SH-SY5Y cells. Mol Biol Rep. 2023;50(1):331-338. doi:10.1007/s11033-022-08039-z
8. 8. Maekawa H, Inagi R. Stress Signal Network between Hypoxia and ER Stress in Chronic Kidney Disease. Front Physiol. 2017;8:74. Published 2017 Feb 8. doi:10.3389/fphys.2017.00074

9. 9. Díaz Bulnes P, Saiz ML, López Larrea C, Rodríguez RM. Crosstalk Between Hypoxia and ER Stress Response: A Key Regulator of Macrophage Polarization. Front Immunol. 2020;10:2951. Published 2020 Jan 8. doi:10.3389/fimmu.2019.02951
10. 10. Zhou D, Li X, Zhao H, et al. Combining multi-dimensional data to identify a key signature (gene and miRNA) of cisplatin-resistant gastric cancer. J Cell Biochem. 2018;119(8):6997-7008. doi:10.1002/jcb.26908
11. 11. Yao X, Li W, Li L, et al. YTHDF1 upregulation mediates hypoxia-dependent breast cancer growth and metastasis through regulating PKM2 to affect glycolysis. Cell Death Dis. 2022;13(3):258. Published 2022 Mar 23. doi:10.1038/s41419-022-04711-1
12. 12. Teles RHG, Moralles HF, Cominetti MR. Global trends in nanomedicine research on triple negative breast cancer: a bibliometric analysis. Int J Nanomedicine. 2018;13:2321-2336. Published 2018 Apr 17. doi:10.2147/IJN.S164355
13. 13. Zhang D, Ye L, Hu S, Zhu Q, Li C, Zhu C. Comprehensive Analysis of the Expression and Prognostic Value of LMAN2 in HER2+ Breast Cancer. J Immunol Res. 2022;2022:7623654. Published 2022 Jun 6. doi:10.1155/2022/7623654
14. 14. Yang G, Höti N, Chen SY, et al. One-Step Enrichment of Intact Glycopeptides From Glycoengineered Chinese Hamster Ovary Cells. Front Chem. 2020;8:240. Published 2020 Apr 17. doi:10.3389/fchem.2020.00240
15. 15. Seagle BL, Eng KH, Yeh JY, et al. Discovery of candidate tumor biomarkers for treatment with intraperitoneal chemotherapy for ovarian cancer. Sci Rep. 2016;6:21591. Published 2016 Feb 17. doi:10.1038/srep21591
16. 16. Marimuthu A, Subbannayya Y, Sahasrabuddhe NA, et al. SILAC-based quantitative proteomic analysis of gastric cancer secretome. Proteomics Clin Appl. 2013;7(5-6):355-366. doi:10.1002/prca.201200069
17. 17. Iurlaro R, Muñoz Pinedo C. Cell death induced by endoplasmic reticulum stress. FEBS J. 2016;283(14):2640-2652. doi:10.1111/febs.13598
18. 18. Isohashi F, Endo H, Mukai M, Inoue T, Inoue M. Insulin-like growth factor stimulation increases radiosensitivity of a pancreatic cancer cell line through endoplasmic reticulum stress under hypoxic conditions. Cancer Sci. 2008;99(12):2395-2401. doi:10.1111/j.1349-7006.2008.00970.x
19. 19. Hori O, Ichinoda F, Yamaguchi A, et al. Role of Herp in the endoplasmic reticulum stress response. Genes Cells. 2004;9(5):457-469. doi:10.1111/j.1356-9597.2004.00735.x