Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression

Hamurcu, Zuhal; AKAR, SAKİNE; Donmez Altuntas, Hamiyet

doi:10.14744/etd.2022.88123

Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression

Sakine Neval AKAR, (Tıbbi Biyoloji Bölümü, Yüzüncü Yıl Üniversitesi Tıp Fakültesi, Van, Türkiye)

Hamiyet Donmez Altuntas, (Tıbbi Biyoloji Bölümü, Erciyes Üniversitesi Tıp Fakültesi, Kayseri, Türkiye)

Zuhal HAMURCU (Tıbbi Biyoloji Bölümü, Erciyes Üniversitesi Tıp Fakültesi, Kayseri, Türkiye)

Erciyes Medical Journal

5 2

Yıl: 2022 Cilt: 44 Sayı: 6 Sayfa Aralığı: 587 - 593 Metin Dili: İngilizce DOI: 10.14744/etd.2022.88123 İndeks Tarihi: 20-05-2023

Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression

Öz:

Objective: Glutamine metabolism is an important pathway in cell proliferation and tumor progression. The first enzyme to be converted in the process of glutamine metabolism, glutaminase 1 (GLS1), exhibits increased expression in many types of cancer, including breast cancer. Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with high glutamine metabolic activity. The aim of this research was to examine the effects on glutamine metabolism and carcinogenic properties following small interfering RNA (siRNA)-mediated inhibition of GLS1 in glutamine-dependent TNBC. Materials and Methods: The effects on cell proliferation, migration, apoptosis, colony formation, and the cell cycle of MDA-MB-231 cells using different siRNAs targeting GLS1 were analyzed using an MTS assay, a wound-healing assay, clo- nogenic analysis, and annexin V and propidium iodide staining methods. The protein expression of GLS1, integrin beta 1 (β1), caspase-3, and p21 were examined using western blot analysis and flow cytometry. Results: The findings revealed that cell viability, migration, and colony formation were significantly suppressed in MDA- MB-231 cells transfected with 2 different GLS1 siRNAs. Furthermore, the results of flow cytometry and western blot analysis demonstrated that knockdown of GLS1 induced arrest in the G0/G1 phase of the cell cycle through the p21 signaling pathway, but did not induce apoptosis. Conclusion: GLS1 is needed for cell proliferation and promotes tumor progression and growth of MDA-MB 231 cells. siRNAs may provide a means to downregulate GLS1 and offer a promising target for breast cancer therapy.

Anahtar Kelime:

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA Cancer J Clin 2019; 69(1): 7–34.
2. Brown JM, Wasson MD, Marcato P. The Missing Lnc: The po - tential of targeting triple-negative breast cancer and cancer stem cells by inhibiting long non-coding RNAs. Cells 2020; 9(3): 763.
3. Pavlova NN, Thompson CB. The emerging hallmarks of cancer metabolism. Cell Metab 2016; 23(1): 27–47.
4. Altman BJ, Stine ZE, Dang CV. From Krebs to clinic: glutamine metabolism to cancer therapy. Nat Rev Cancer 2016; 16(10): 619–34.
5. Yu W, Yang X, Zhang Q, Sun L, Yuan S, Xin Y. Targeting GLS1 to cancer therapy through glutamine metabolism. Clin Transl Oncol 2021; 23(11): 2253–68.
6. Gross MI, Demo SD, Dennison JB, Chen L, Chernov-Rogan T, Goyal B, et al. Antitumor activity of the glutaminase inhib - itor CB-839 in triple-negative breast cancer. Mol Cancer Ther 2014; 13(4): 890–901.
7. Huang F, Zhang Q, Ma H, Lv Q, Zhang T. Expression of gluta- minase is upregulated in colorectal cancer and of clinical signifi- cance. Int J Clin Exp Pathol 2014; 7(3): 1093–100.
8. Xi J, Sun Y, Zhang M, Fa Z, Wan Y, Min Z, et al. GLS1 pro- motes proliferation in hepatocellular carcinoma cells via AKT/ GSK3β/CyclinD1 pathway. Exp Cell Res 2019; 381(1): 1–9.
9. Pan T, Gao L, Wu G, Shen G, Xie S, Wen H, et al. Elevated expression of glutaminase confers glucose utilization via gluta- minolysis in prostate cancer. Biochem Biophys Res Commun 2015; 456(1): 452–8.
10. Matre P, Velez J, Jacamo R, Qi Y, Su X, Cai T, et al. Inhibiting glutaminase in acute myeloid leukemia: metabolic dependency of selected AML subtypes. Oncotarget 2016; 7(48): 79722–35.
11. Wang D, Meng G, Zheng M, Zhang Y, Chen A, Wu J, et al. The Glutaminase-1 Inhibitor 968 enhances dihydroartemisinin-medi- ated antitumor efficacy in hepatocellular carcinoma cells. PLoS One 2016; 11(11): e0166423.
12. Yang J, Guo Y, Seo W, Zhang R, Lu C, Wang Y, et al. Targeting cellular metabolism to reduce head and neck cancer growth. Sci Rep 2019; 9(1): 4995.
13. Kim S, Kim DH, Jung WH, Koo JS. Expression of glutamine metabolism-related proteins according to molecular subtype of breast cancer. Endocr Relat Cancer 2013; 20(3): 339–48.
14. van Geldermalsen M, Wang Q, Nagarajah R, Marshall AD, Thoeng A, Gao D, et al. ASCT2/SLC1A5 controls glutamine uptake and tumour growth in triple-negative basal-like breast cancer. Oncogene 2016; 35(24): 3201–8.
15. Hamurcu Z, Delibaşı N, Geçene S, Şener EF, Dцnmez-Altun- taş H, Özkul Y, et al. Targeting LC3 and Beclin-1 autophagy genes suppresses proliferation, survival, migration and invasion by inhibition of Cyclin-D1 and uPAR/Integrin β1/ Src signaling in triple negative breast cancer cells. J Cancer Res Clin Oncol 2018; 144(3): 415–30.
16. Hamurcu Z, Delibaşı N, Nalbantoglu U, Sener EF, Nurdinov N, Tascı B, et al. FOXM1 plays a role in autophagy by tran - scriptionally regulating Beclin-1 and LC3 genes in human tri- ple-negative breast cancer cells. J Mol Med (Berl) 2019; 97(4): 491–508.
17. Jin H, Varner J. Integrins: roles in cancer development and as treatment targets. Br J Cancer 2004; 90(3): 561–5.
18. Julien O, Wells JA. Caspases and their substrates. Cell Death Differ 2017; 24(8): 1380–9.
19. Bertoli C, Skotheim JM, de Bruin RA. Control of cell cycle tran- scription during G1 and S phases. Nat Rev Mol Cell Biol 2013; 14(8): 518–28.
20. Szeliga M, Bogacińska-Karaś M, Różycka A, Hilgier W, Marquez J, Albrecht J. Silencing of GLS and overexpression of GLS2 genes cooperate in decreasing the proliferation and viability of glioblastoma cells. Tumour Biol 2014; 35(3): 1855–62.
21. Jacque N, Ronchetti AM, Larrue C, Meunier G, Birsen R, Wil- lems L, et al. Targeting glutaminolysis has antileukemic activity in acute myeloid leukemia and synergizes with BCL-2 inhibition. Blood 2015; 126(11): 1346–56.
22. Ye X, Zhou Q, Matsumoto Y, Moriyama M, Kageyama S, Komatsu M, et al. Inhibition of glutaminolysis ınhibits cell growth via down-regulating Mtorc1 signaling in lung squamous cell carcinoma. Anticancer Res 2016; 36(11): 6021–9.
23. Yuan L, Sheng X, Willson AK, Roque DR, Stine JE, Guo H, et al. Glutamine promotes ovarian cancer cell proliferation through the mTOR/S6 pathway. Endocr Relat Cancer 2015; 22(4): 577–91.
24. Yuan L, Sheng X, Clark LH, Zhang L, Guo H, Jones HM, et al. Glutaminase inhibitor compound 968 inhibits cell prolifera- tion and sensitizes paclitaxel in ovarian cancer. Am J Transl Res 2016; 8(10): 4265–77.
25. Lampa M, Arlt H, He T, Ospina B, Reeves J, Zhang B, et al. Glutaminase is essential for the growth of triple-negative breast cancer cells with a deregulated glutamine metabolism pathway and its suppression synergizes with mTOR inhibition. PLoS One 2017; 12(9): e0185092.
26. Han T, Guo M, Zhang T, Gan M, Xie C, Wang JB. A novel glutaminase inhibitor-968 inhibits the migration and prolifera- tion of non-small cell lung cancer cells by targeting EGFR/ERK signaling pathway. Oncotarget 2017; 8(17): 28063–73.
27. Lu WQ, Hu YY, Lin XP, Fan W. Knockdown of PKM2 and GLS1 expression can significantly reverse oxaliplatin-resistance in colorectal cancer cells. Oncotarget 2017; 8(27): 44171–85.
28. Masamha CP, LaFontaine P. Molecular targeting of glutaminase sensitizes ovarian cancer cells to chemotherapy. J Cell Biochem 2018; 119(7): 6136–45.
29. Li B, Cao Y, Meng G, Qian L, Xu T, Yan C, et al. Targeting glu- taminase 1 attenuates stemness properties in hepatocellular car- cinoma by increasing reactive oxygen species and suppressing Wnt/beta-catenin pathway. EBioMedicine 2019; 39: 239–54.
30. Liu HY, Zhang HS, Liu MY, Li HM, Wang XY, Wang M. GLS1 depletion inhibited colorectal cancer proliferation and migration via redox/Nrf2/autophagy-dependent pathway. Arch Biochem Biophys 2021; 708: 108964.

APA	AKAR S, Donmez Altuntas H, Hamurcu Z (2022). Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression. , 587 - 593. 10.14744/etd.2022.88123
Chicago	AKAR SAKİNE,Donmez Altuntas Hamiyet,Hamurcu Zuhal Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression. (2022): 587 - 593. 10.14744/etd.2022.88123
MLA	AKAR SAKİNE,Donmez Altuntas Hamiyet,Hamurcu Zuhal Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression. , 2022, ss.587 - 593. 10.14744/etd.2022.88123
AMA	AKAR S,Donmez Altuntas H,Hamurcu Z Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression. . 2022; 587 - 593. 10.14744/etd.2022.88123
Vancouver	AKAR S,Donmez Altuntas H,Hamurcu Z Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression. . 2022; 587 - 593. 10.14744/etd.2022.88123
IEEE	AKAR S,Donmez Altuntas H,Hamurcu Z "Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression." , ss.587 - 593, 2022. 10.14744/etd.2022.88123
ISNAD	AKAR, SAKİNE vd. "Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression". (2022), 587-593. https://doi.org/10.14744/etd.2022.88123

APA	AKAR S, Donmez Altuntas H, Hamurcu Z (2022). Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression. Erciyes Medical Journal, 44(6), 587 - 593. 10.14744/etd.2022.88123
Chicago	AKAR SAKİNE,Donmez Altuntas Hamiyet,Hamurcu Zuhal Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression. Erciyes Medical Journal 44, no.6 (2022): 587 - 593. 10.14744/etd.2022.88123
MLA	AKAR SAKİNE,Donmez Altuntas Hamiyet,Hamurcu Zuhal Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression. Erciyes Medical Journal, vol.44, no.6, 2022, ss.587 - 593. 10.14744/etd.2022.88123
AMA	AKAR S,Donmez Altuntas H,Hamurcu Z Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression. Erciyes Medical Journal. 2022; 44(6): 587 - 593. 10.14744/etd.2022.88123
Vancouver	AKAR S,Donmez Altuntas H,Hamurcu Z Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression. Erciyes Medical Journal. 2022; 44(6): 587 - 593. 10.14744/etd.2022.88123
IEEE	AKAR S,Donmez Altuntas H,Hamurcu Z "Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression." Erciyes Medical Journal, 44, ss.587 - 593, 2022. 10.14744/etd.2022.88123
ISNAD	AKAR, SAKİNE vd. "Downregulation of glutaminase 1 (GLS1) Inhibits Proliferation, Clonogenicity, and Migration of Aggressive MDA-MB-231 Breast Cancer Cells by Increasing p21 and Decreasing Integrin-β1 Expression". Erciyes Medical Journal 44/6 (2022), 587-593. https://doi.org/10.14744/etd.2022.88123