Inhibition of NOTCH1 signaling in tumor-initiating cells overcomes chemoresistance and promotes apoptosis

Authors

  • Yang Rui Department of General Surgery, Central Theater Command General Hospital of PLA, Wuhan, 430000, PR China
  • Zhan Gang Department of General Surgery, Central Theater Command General Hospital of PLA, Wuhan, 430000, PR China
  • Zhou Jun Department of General Surgery, Central Theater Command General Hospital of PLA, Wuhan, 430000, PR China
  • Jin Weidong Department of General Surgery, Central Theater Command General Hospital of PLA, Wuhan, 430000, PR China
  • Jiang Hui Department of General Surgery, Central Theater Command General Hospital of PLA, Wuhan, 430000, PR China

DOI:

https://doi.org/10.2298/ABS210423030R

Keywords:

apoptosis, drug resistance, hepatocellular carcinoma, NOTCH1, tumor-initiating cells

Abstract

Paper description:

  • Aberrant upregulation of Notch1 in hepatocellular cancer (HCC) attenuates apoptosis.
  • The role of the Notch1 signaling pathway in apoptosis inhibition in liver cancer cell lines was investigated.
  • After silencing Notch1 expression by siRNA transfection of tumor initiating cells, the intrinsic apoptotic pathway was induced through downregulation of the PI3/AKT signaling pathway and the cells became sensitive to chemotherapeutic drug treatment.
  • Pharmacological inactivation of Notch1 presents a potentially attractive clinical target for treating HCC.

Abstract: The Notch signaling pathway is an evolutionarily conserved pathway essential for regulation of cell development and differentiation. Upregulation and activation of Notch signaling enhances the oncogenic potential of cancer cells through apoptosis resistance. The NOTCH1 expression pattern in hepatocellular cancer (HCC) and its role in apoptosis attenuation was determined. Immunohistostaining identified intensive positive staining of NOTCH1 in human HCC tissues as compared to control tissues. RT-PCR and Western blot quantification data showed that NOTCH1 and its downstream target transcription factor Hes1 were significantly upregulated in HCC cells. Based on these findings, we separated a population of CD44+ tumor-initiating cells (HepG2: >7%; SNU449: >6%) from HCC cell lines to ascertain the role of NOTCH1 in tumorigenesis. After NOTCH1-specific small interfering RNA (siRNA) transfection of tumor-initiating cells (TICs), NOTCH1 was significantly downregulated, and efficient uptake of DNA-targeting chemotherapeutic drugs was observed. Meanwhile, by flow cytometry analysis we found that the rate of apoptosis induction was significantly higher (P<0.01) and that cell viability was reduced (HepG2<23%; SNU449<28%) in siRNA transfected cells. In addition, the release of cytochrome C and activation of caspase 9 in CD44+ TICs was observed after siRNA transfection, confirming the induction of the mitochondrial-dependent intrinsic apoptotic pathway. Western blot analysis revealed inhibition of the PI3-Akt signaling pathway in siRNA-transfected TICs. These data suggest that activated NOTCH1 plays a significant role in liver cancer progression through apoptosis inhibition via regulation of PI3-Akt signaling. Therefore, pharmacological inactivation of NOTCH1 represents a clinically relevant therapeutic target for treating HCC.

Downloads

Download data is not yet available.

References

Jue C, Lin C, Zhisheng Z, Yayun Q, Feng J, Min Z, Haibo W, Youyang S, Hisamitsu T, Shintaro I, Shiyu G. Notch1 promotes vasculogenic mimicry in hepatocellular carcinoma by inducing EMT signaling. Oncotarget. 2017;8(2):2501. https://doi.org/10.18632/oncotarget.12388

Abou-Alfa GK, Venook AP. The antiangiogenic ceiling in hepatocellular carcinoma: does it exist and has it been reached? Lancet Oncol. 2013;14:e283-8. https://doi.org/10.1016/S1470-2045(13)70161-X

Shinoda M, Kishida N, Itano O, Ei S, Ueno A, Kitago M, Abe Y, Hibi T, Yagi H, Masugi Y, Tanabe M. Long-term complete response of advanced hepatocellular carcinoma treated with multidisciplinary therapy including reduced dose of sorafenib: case report and review of the literature. World J Surg Oncol. 2015;13:144. https://doi.org/10.1186/s12957-015-0559-9

Guo W, He X, Li Z, Li Y. Combination of transarterial chemoembolization (tace) and radiofrequency ablation (rfa) vs. surgical resection (sr) on survival outcome of early hepatocellular carcinoma: A meta-analysis. Hepatogastroenterology. 2015;62:710-4.

Matsui W, Huff CA, Wang Q, Malehorn MT, Barber J, Tanhehco Y, Smith BD, Civin CI, Jones RJ. Characterization of clonogenic multiple myeloma cells. Blood. 2004;103(6):2332-6. https://doi.org/10.1182/blood-2003-09-3064

Prince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P, Weissman IL, Clarke MF, Ailles LE. Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci U S A. 2007;104(3):973-8. https://doi.org/10.1073/pnas.0610117104

Naor D, Wallach-Dayan SB, Zahalka MA, Sionov RV. Involvement of CD44, a molecule with a thousand faces, in cancer dissemination. Semin Cancer Biol. 2008;18(4):260-67. https://doi.org/10.1016/j.semcancer.2008.03.015

Jaggupilli A, Elkord E. Significance of CD44 and CD24 as cancer stem cell markers: an enduring ambiguity. Clin Dev Immunol. 2012;2012:708036. https://doi.org/10.1155/2012/708036

Chung J, Riella LV, Maillard I. Targeting the Notch pathway to prevent rejection. Am J Transplant. 2016;16:3079-85. https://doi.org/10.1111/ajt.13816

Pancewicz-Wojtkiewicz J. Epidermal growth factor receptor and notch signaling in non-small-cell lung cancer. Cancer Med. 2016;5:3572-78. https://doi.org/10.1002/cam4.944

Sui C, Zhuang C, Sun D, Yang L, Zhang L, Song L. Notch1 regulates the JNK signaling pathway and increases apoptosis in hepatocellular carcinoma. Oncotarget. 2017;8(28):45837. https://doi.org/10.18632/oncotarget.17434

Ning LI, Wentworth L, Chen H, Weber SM. Down-regulation of Notch1 signaling inhibits tumor growth in human hepatocellular carcinoma. Am J Transl Res. 2009;1(4):358.

Sun T, Zhang D, Wang Z, Zhao B, Li Y, Sun X, Liu J, Wang X, Sheng J. Inhibition of the notch signaling pathway overcomes resistance of cervical cancer cells to paclitaxel through retardation of the epithelial-mesenchymal transition process. Environ Toxicol. 2021;https://doi.org/10.1002/tox.23296. https://doi.org/10.1002/tox.23296

Purow B. Notch inhibition as a promising new approach to cancer therapy. Adv Exp Med Biol. 2012;727:305-19. https://doi.org/10.1007/978-1-4614-0899-4_23

Gu Y, Masiero M and Banham AH. Notch signaling: its roles and therapeutic potential in hematological malignancies. Oncotarget. 2016;7:29804-23. https://doi.org/10.18632/oncotarget.7772

He QZ, Luo XZ, Wang K, Zhou Q, Ao H, Yang Y, Li SX, Li Y, Zhu HT, Duan T. Isolation and Characterization of Cancer Stem Cells from High-Grade Serous Ovarian Carcinomas. Cell Physiol Biochem. 2014;33(1):173-84. https://doi.org/10.1159/000356660

McIlwain DR, Berger Tand Mak TW. Caspase functions in cell death and disease. Cold Spring Harb Perspect Biol. 2015;7(4):a026716. https://doi.org/10.1101/cshperspect.a026716

Elena-Real CA, Díaz-Quintana A, González-Arzola K, Velázquez-Campoy A, Orzáez M, López-Rivas A, Gil-Caballero S, Miguel Á, Díaz-Moreno I. Cytochrome c speeds up caspase cascade activation by blocking 14-3-3ε-dependent Apaf-1 inhibition. Cell Death Dis. 2018;9(3):1-2. https://doi.org/10.1038/s41419-018-0408-1

Allenspach EJ, Maillard I, Aster JC, Pear WS. Notch signaling in cancer. Cancer Biol.Ther. 2002;1:466-76. https://doi.org/10.4161/cbt.1.5.159

Miele L. Notch signaling. Clin Cancer Res. 2006;12:1074-9. https://doi.org/10.1158/1078-0432.CCR-05-2570

Miele L, Miao H, Nickoloff BJ. NOTCH signaling as a novel cancer therapeutic target. Curr Cancer Drug Targets. 2006;6:313-23. https://doi.org/10.2174/156800906777441771

Giovannini C, Minguzzi M, Genovese F, Baglioni M, Gualandi A, Ravaioli M, Milazzo M, Tavolari S, Bolondi L, Gramantieri L. Molecular and proteomic insight into Notch1 characterization in hepatocellular carcinoma. Oncotarget. 2016;7(26):39609. https://doi.org/10.18632/oncotarget.9203

Du L, Wang H, He L, Zhang J, Ni B, Wang X, Jin H, Cahuzac N, Mehrpour M, Lu Y, Chen Q. CD44 is of functional importance for colorectal cancer stem cells. Clin Cancer Res. 2008;14(21):6751-60. https://doi.org/10.1158/1078-0432.CCR-08-1034

Villanueva A, Alsinet C, Yanger K, Hoshida Y, Zong Y, Toffanin S, Rodriguez-Carunchio L, Solé M, Thung S, Stanger BZ, Llovet JM. Notch signaling is activated in human hepatocellular carcinoma and induces tumor formation in mice. Gastroenterology. 2012;143:1660-1669e7. https://doi.org/10.1053/j.gastro.2012.09.002

Han B, Liu SH, Guo WD, Zhang B, Wang JP, Cao YK, Liu J. Notch1 downregulation combined with interleukin-24 inhibits invasion and migration of hepatocellular carcinoma cells. World J Gastroenterol. 2015;21:9727-35. https://doi.org/10.3748/wjg.v21.i33.9727

Huntzicker EG, Hotzel K, Choy L, Che L, Ross J, Pau G, Sharma N, Siebel CW, Chen X, French DM. Differential effects of targeting Notch receptors in a mouse model of liver cancer. Hepatology. 2015;61:942-52. https://doi.org/10.1002/hep.27566

Geisler F, Strazzabosco M. Emerging roles of Notch signaling in liver disease. Hepatology. 2015;61:382-92. https://doi.org/10.1002/hep.27268

Downloads

Published

2021-10-12

How to Cite

1.
Rui Y, Gang Z, Jun Z, Weidong J, Hui J. Inhibition of NOTCH1 signaling in tumor-initiating cells overcomes chemoresistance and promotes apoptosis. Arch Biol Sci [Internet]. 2021Oct.12 [cited 2021Dec.8];73(3):371-9. Available from: https://serbiosoc.org.rs/arch/index.php/abs/article/view/6541

Issue

Section

Articles