Mechanism of Notch1‑saRNA‑1480 reversing androgen sensitivity in human metastatic castration‑resistant prostate cancer

Ma,Libin; Jiang,Kang; Jiang,Peiwu; He,Han; Chen,Kean; Shao,Jia; Deng,Gang

doi:10.3892/ijmm.2020.4597

Mechanism of Notch1‑saRNA‑1480 reversing androgen sensitivity in human metastatic castration‑resistant prostate cancer

Authors:
- Libin Ma
- Kang Jiang
- Peiwu Jiang
- Han He
- Kean Chen
- Jia Shao
- Gang Deng
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Affiliations: Department of Nephrology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310016, P.R. China, Department of Urology, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, P.R. China, Surgical Department Ⅰ, Hangzhou Hospital of Traditional Chinese Medicine, Hangzhou, Zhejiang 310007, P.R. China
Published online on: May 8, 2020 https://doi.org/10.3892/ijmm.2020.4597
Pages: 265-279
Copyright: © Ma et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to explore the mechanism by which Notch1‑small activating (sa)RNA restored androgen sensitivity in human metastatic castration‑resistant prostate cancer (CRPC). After transfection of Notch1‑saRNA‑1480 in PC3 cells, the expression of Notch1 and androgen receptor (AR) was investigated by reverse transcription quantitative PCR (RT‑qPCR) and western blotting. Furthermore, the protein expression level of vascular endothelial growth factor (VEGF) was measured. Then, flow cytometry was used to analyze the cell cycle and apoptosis after transfection. Moreover, the migration and invasion ability of PC3 cells were assessed by transwell assays. Then, angiogenesis experiments were conducted to analyze the abilities of PC3 cells to form blood vessels. Furthermore, in vivo experiments detected the antitumor activity of Notch1‑saRNA‑1480. The mRNA and protein expression levels of Notch1 were significantly increased after transfection, while the expression levels of AR and VEGF were decreased. After transfection, the cell cycle was arrested at the G0/G1 checkpoint. Notch1‑saRNA‑1480 significantly increased the proportion of apoptotic cells after transfection. In addition, transwell assay results showed that PC3 cell migration and invasion were inhibited. The total vessel length was significantly decreased based on angiogenesis experiments, which indicated that PC3 cell angiogenesis was inhibited. In vivo experiments showed that Notch1‑saRNA‑1480 could inhibit tumor growth and volume. The protein expression of Notch1, AR, VEGF receptor 2 (VEGFR2) and VEGF in tumor tissues was consistent with in vitro levels. Notch1‑saRNA‑1480 could significantly inhibit the proliferation of PC3 cells in vitro and the growth of tumors in vivo, which is associated with the inhibition of the AR and VEGF pathways.

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1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2017. CA Cancer J Clin. 67:7–30. 2017. View Article : Google Scholar : PubMed/NCBI
2	Filson CP, Marks LS and Litwin MS: Expectant management for men with early stage prostate cancer. CA Cancer J Clin. 65:265–282. 2015. View Article : Google Scholar : PubMed/NCBI
3	Chen WY, Tsai YC, Yeh HL, Suau F, Jiang KC, Shao AN, Huang J and Liu YN: Loss of SPDEF and gain of TGFBI activity after androgen deprivation therapy promote EMT and bone metastasis of prostate cancer. Sci Signal. 10:eaam68262017. View Article : Google Scholar : PubMed/NCBI
4	Stoyanova T, Riedinger M, Lin S, Faltermeier CM, Smith BA, Zhang KX, Going CC, Goldstein AS, Lee JK, Drake JM, et al: Activation of Notch1 synergizes with multiple pathways in promoting castration-resistant prostate cancer. Proc Natl Acad Sci USA. 113:E6457–E6466. 2016. View Article : Google Scholar : PubMed/NCBI
5	Carvalho FL, Simons BW, Eberhart CG and Berman DM: Notch signaling in prostate cancer: A moving target. Prostate. 74:933–945. 2014. View Article : Google Scholar : PubMed/NCBI
6	Deng G, Ma L, Meng Q, Ju X, Jiang K, Jiang P and Yu Z: Notch signaling in the prostate: Critical roles during development and in the hallmarks of prostate cancer biology. J Cancer Res Clin Oncol. 142:531–547. 2016. View Article : Google Scholar
7	Nabbi A, McClurg UL, Thalappilly S, Almami A, Mobahat M, Bismar TA, Binda O and Riabowol KT: ING3 promotes prostate cancer growth by activating the androgen receptor. BMC Med. 15:1032017. View Article : Google Scholar : PubMed/NCBI
8	Miyamoto H, Meming EM and Chang C: Androgen deprivation therapy for prostate cancer: Current status and future prospects. Prostate. 61:332–353. 2004. View Article : Google Scholar : PubMed/NCBI
9	Stanbrough M, Bubley GJ, Ross K, Golub TR, Rubin MA, Penning TM, Febbo PG and Balk SP: Increased expression of genes converting adrenal androgens to testosterone in androgen-independent prostate cancer. Cancer Res. 66:2815–2825. 2006. View Article : Google Scholar : PubMed/NCBI
10	Wang Y, Wu X, Ou L, Yang X, Wang X, Tang M, Chen E and Luo C: PLCε knockdown inhibits prostate cancer cell proliferation via suppression of Notch signalling and nuclear translocation of the androgen receptor. Cancer Lett. 362:61–69. 2015. View Article : Google Scholar : PubMed/NCBI
11	Belandia B, Powell SM, García-Pedrero JM, Walker MM, Bevan CL and Parker MG: Hey1, a mediator of notch signaling, is an androgen receptor corepressor. Mol Cell Biol. 25:1425–1436. 2005. View Article : Google Scholar : PubMed/NCBI
12	Sadick H, Naim R, Gössler U, Hörmann K and Riedel F: Angiogenesis in hereditary hemorrhagic telangiectasia: VEGF165 plasma concentration in correlation to the VEGF expression and microvessel density. Int J Mol Med. 15:15–19. 2005.
13	Shou J, Ross S, Koeppen H, de Sauvage FJ and Gao WQ: Dynamics of notch expression during murine prostate development and tumorigenesis. Cancer Res. 61:7291–7297. 2001.PubMed/NCBI
14	D'Amore PA and Ng YS: Won't you be my neighbor? Local induction of arteriogenesis. Cell. 110:289–292. 2002. View Article : Google Scholar : PubMed/NCBI
15	Siekmann AF, Covassin L and Lawson ND: Modulation of VEGF signalling output by the Notch pathway. Bioessays. 30:303–313. 2008. View Article : Google Scholar : PubMed/NCBI
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
17	Roberts TC, Hart JR, Kaikkonen MU, Weinberg MS, Vogt PK and Morris KV: Quantification of nascent transcription by bromouridine immunocapture nuclear run-on RT-qPCR. Nat Protoc. 10:1198–1211. 2015. View Article : Google Scholar : PubMed/NCBI
18	Alimirah F1, Chen J, Basrawala Z, Xin H and Choubey D: DU-145 and PC-3 human prostate cancer cell lines express androgen receptor: Implications for the androgen receptor functions and regulation. FEBS Lett. 580:2294–2300. 2006. View Article : Google Scholar : PubMed/NCBI
19	Davis R, Jia D, Cinar B, Sikka SC, Moparty K, Zhau HE, Chung LW, Agrawal KC and Abdel-Mageed AB: Functional androgen receptor confers sensitization of androgen-independent prostate cancer cells to anticancer therapy via caspase activation. Biochem Biophys Res Commun. 309:937–945. 2003. View Article : Google Scholar : PubMed/NCBI
20	Deng G, Ju X, Meng Q, Yu ZJ and Ma LB: Emodin inhibits the proliferation of PC3 prostate cancer cells in vitro via the Notch signaling pathway. Mol Med Rep. 12:4427–4433. 2015. View Article : Google Scholar : PubMed/NCBI
21	Zhang L, Sha J, Yang G, Huang X, Bo J and Huang Y: Activation of Notch pathway is linked with epithelial-mesenchymal transition in prostate cancer cells. Cell Cycle. 16:999–1007. 2017. View Article : Google Scholar : PubMed/NCBI
22	Yang JH, Wylie-Sears J and Bischoff J: Opposing actions of Notch1 and VEGF in post-natal cardiac valve endothelial cells. Biochem Biophys Res Commun. 374:512–516. 2008. View Article : Google Scholar : PubMed/NCBI
23	Bao B, Wang Z, Ali S, Kong D, Li Y, Ahmad A, Banerjee S, Azmi AS, Miele L and Sarkar FH: Notch-1 induces epithelial-mesenchymal transition consistent with cancer stem cell phenotype in pancreatic cancer cells. Cancer Lett. 307:26–36. 2011. View Article : Google Scholar : PubMed/NCBI
24	Mao Q, Zheng X, Yang K, Qin J, Bai Y, Jia X, Li Y and Xie L: Suppression of migration and invasion of PC3 prostate cancer cell line via activating E-cadherin expression by small activating RNA. Cancer Invest. 28:1013–1018. 2010. View Article : Google Scholar : PubMed/NCBI
25	Yang K, Zheng XY, Qin J, Wang YB, Bai Y, Mao QQ, Wan Q, Wu ZM and Xie LP: Up-regulation of p21WAF1/Cip1 by saRNA induces G1-phase arrest and apoptosis in T24 human bladder cancer cells. Cancer Lett. 265:206–214. 2008. View Article : Google Scholar : PubMed/NCBI
26	Liu ZJ, Shirakawa T, Li Y, Soma A, Oka M, Dotto GP, Fairman RM, Velazquez OC and Herlyn M: Regulation of Notch1 and Dll4 by vascular endothelial growth factor in arterial endothelial cells: Implications for modulating arteriogenesis and angiogenesis. Mol Cell Biol. 23:14–25. 2003. View Article : Google Scholar :
27	Larrivée B, Prahst C, Gordon E, del Toro R, Mathivet T, Duarte A, Simons M and Eichmann A: ALK1 signaling inhibits angiogenesis by cooperating with the Notch pathway. Dev Cell. 22:489–500. 2012. View Article : Google Scholar : PubMed/NCBI
28	Yang J, Wang C, Zhang Z, Chen X, Jia Y, Wang B and Kong T: Curcumin inhibits the survival and metastasis of prostate cancer cells via the Notch-1 signaling pathway. APMIS. 125:134–140. 2017. View Article : Google Scholar : PubMed/NCBI
29	Sahara M, Hansson EM, Wernet O, Lui KO, Später D and Chien KR: Manipulation of a VEGF-Notch signaling circuit drives formation of functional vascular endothelial progenitors from human pluripotent stem cells. Cell Res. 25:1482015. View Article : Google Scholar : PubMed/NCBI
30	Zang M, Hu L, Zhang B, Zhu Z, Li J, Zhu Z, Yan M and Liu B: Luteolin suppresses angiogenesis and vasculogenic mimicry formation through inhibiting Notch1-VEGF signaling in gastric cancer. Biochem Biophys Res Commun. 490:913–919. 2017. View Article : Google Scholar : PubMed/NCBI
31	Esquela-Kerscher A and Slack FJ: Oncomirs-microRNAs with a role in cancer. Nat Rev Cancer. 6:259–269. 2006. View Article : Google Scholar : PubMed/NCBI
32	Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K and Tuschl T: Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature. 411:494–498. 2001. View Article : Google Scholar : PubMed/NCBI
33	Chen Z, Place RF, Jia ZJ, Pookot D, Dahiya R and Li LC: Antitumor effect of dsRNA-induced p21(WAF1/CIP1) gene activation in human bladder cancer cells. Mol Cancer Ther. 7:698–703. 2008. View Article : Google Scholar : PubMed/NCBI