Effect of silencing Notch1 and Notch2 on the expression of Hes family genes in HPAC pancreatic cancer cells

doi:10.3969/j.issn.1006-7795.2015.02.015

Abstract

Abstract: Objective To explore the effect of silencing Notch1 and Notch2 on the gene expression of Hes family target genes in HPAC pancreatic cancer cells. Methods Notch1-siRNA and Notch2-siRNA were transfected as a liposomal formulation into HPAC cells, respectively, the protein expression of Notch1 and Notch2 were detected by Western blotting, and the mRNA levels of Hes1, Hes2, Hes4 and Hes6 were detected by real-time PCR, and the protein expression level of Hes1 was detected by Western blotting. Results The protein level of Notch1 in Notch1-siRNA group (0.124±0.005) was significantly decreased compared to that of the blank group (1.000±0.019) and RNAi control group (0.908±0.039). Down-regulation of Notch1 expression decreased the mRNA expression of Hes1 and Hes6(Hes1 vs that of the blank group, P=0.025, Hes1 vs RNAi control group, P=0.024; Hes6 vs the blank group, P=0.018, Hes6 vs RNAi control group, P=0.012), but has no effect on the expression of Hes2 and Hes4 mRNA(Hes2 vs the blank group, P=0.064, Hes2 vs RNAi control group, P=0.059; Hes4 vs the blank group, P=0.069, Hes4 vs RNAi control group, P=0.071. The protein expression of Notch2 in Notch2-siRNA group(0.350±0.009) was significantly decreased, compared to the blank group (1.000±0.015) and RNAi control group(0.990±0.017). Down-regulation of Notch2 decreased the mRNA expression of Hes1(Hes1 vs the blank group, P=0.015, Hes1 vs RNAi control group, P=0.014), but has no effect on the expression of Hes2, Hes4 and Hes6(Hes2 vs the blank group, P=0.178; Hes2 vs RNAi control group, P=0.121; Hes4 vs the blank group, P=0.097, Hes6 vs RNAi control group, P=0.100; Hes6 vs the blank group, P=0.053, Hes6 vs RNAi control group, P=0.088. Neither of down-regulation of Notch1 and Notch2 down-regulate the protein expression of Hes1. Conclusion The downstream target gene of Notch1 is Hes1 and Hes6, and the downstream target gene of Notch2 is Hes1. Our results indicate that Notch1 and Notch2 have different regulatory effects on the expressions of Hes family genes.

Key words: Notch1, Notch2, siRNA, Hes family genes, pancreatic cancer

CLC Number:

R329.2

Li Bingqiu, Zhang Yuxiang. Effect of silencing Notch1 and Notch2 on the expression of Hes family genes in HPAC pancreatic cancer cells[J]. Journal of Capital Medical University, 2015, 36(2): 244-250.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2015.02.015

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2015/V36/I2/244

References

[1] Gordon W R, Arnett K L, Blacklow S C. The molecular logic of Notch signaling-a structural and biochemical perspective[J]. J Cell Sci, 2008,121(19):3109-3119.
[2] Miyamoto Y, Maitra A, Ghosh B, et al. Notch mediates TGF alpha induced changes in epithelial differentiation during pancreatictumorigenesis[J]. Cancer Cell, 2003,3(6):565-576.
[3] Bao B, Wang Z W, Ali S, et al. Notch-1 induces epithelial-mesenchymal transition consistent with cancer stem cell phenotype in pancreatic cancer cells[J]. Cancer Lett, 2011,307(1):26-36.
[4] Wang Z W, Zhang Y X, Li Y W, et al. Down-regulation of Notch-1 contributes to cell growth inhibition and apoptosis in pancreatic cancer cells[J]. Mol Cancer Ther, 2006,5(3):483-493.
[5] Avila J L, Kissil J L. Notch signaling in pancreatic cancer:oncogene or tumor suppressor?[J]. Cell Press, 2013,19(5):320-327.
[6] Fiuza U M, Arias A M. Cell and molecular biology of Notch[J]. J Endocrinol, 2007,194(3):459-474.
[7] Roy M, Pear W S, Aster J C. The multifaceted role of Notch in cancer[J]. Curr Opin Genet Dev, 2007,17(1):52-59.
[8] Kopan R, Ilagan M X. The canonical Notch signaling pathway:unfolding the activation mechanism[J]. Cell, 2009,137(2):216-233.
[9] 常思佳,邹晓英,庄姮,等.激活Notch信号通路可延迟人牙髓细胞的老化表现[J].北京大学学报:医学版,2014,46(1):5-11.
[10] Bai G, Cheung I, Shulha H P, et al. Epigenetic dysregulation of hairy and enhancer of split 4(HES4) is associated with striatal degeneration in postmortem Huntington brains[J]. Hum Mol Genet, 2014,24(5):1441-1456.
[11] Wickramasinghe C M, Domaschenz R, Amagase Y, et al. HES6 enhances the motility of alveolar rhabdomyosarcoma cells[J]. Exp Cell Res, 2013,319(1):103-112.
[12] Abel E V, Kim E J, Wu J J, et al. The Notch pathway is important in maintaining the cancer stem cell population in pancreatic cancer[J]. PLoS One, 2014,9(3):e91983.
[13] 朱雷,盛伟伟,董明,等.Notch1在胰腺癌中的表达及临床病理学意义[J].中国医科大学学报,2014,43(4):320-323.
[14] Wharton K A, Johansen K M, Xu T, et al. Nucleotide sequence from the neurogenic locus notch implies a gene product that shares homology with proteins containing EGF-like repeats[J]. Cell, 1985,43(3 Pt 2):567-581.
[15] Mazur P K, Einwächter H, Lee M, et al. Notch2 is required for progression of pancreatic intraepithelial neoplasia and development of pancreatic ductal adenocarcinoma[J]. Proc Natl Acad Sci, 2010,107:13438-13443.
[16] Yuan Z, Friedmann D R, VanderWielen B D, et al. Characterization of CSL(CBF-1, Su(H), Lag-1) mutants reveals differences in signaling mediated by Notch1 and Notch2[J]. J Biol Chem, 2012,287(42):34904-34916.
[17] Yoshiura S, Ohtsuka T, Takenaka Y, et al. Ultradian oscillations of Stat, Smad, and Hes1 expression in response to serum?[J]. Proc Natl Acad Sci, 2007,104(27):11292-11297.