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14.07.2022 | Research

Improved delivery of Mcl-1 and survivin siRNA combination in breast cancer cells with additive siRNA complexes

verfasst von: Tinnabhop Santadkha, Wanwisa Skolpap, Remant K.C., Aysha Ansari, Cezary Kucharski, Teo Atz Dick, Hasan Uludağ

Erschienen in: Investigational New Drugs | Ausgabe 5/2022

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Summary

This study aimed at investigating the influence of commercial transfection reagents (Prime-Fect, Leu-Fect A, and Leu-Fect C) complexed with different siRNAs (CDC20, HSP90, Mcl-1 and Survivin) in MDA-MB-436 breast cancer cells and the impact of incorporating an anionic additive, Trans-Booster, into siRNA formulations for improving in vitro gene silencing and delivery efficiency. Gene silencing was quantitatively analyzed by real-time RT-PCR while cell proliferation and siRNA uptake were evaluated by the MTT assay and flow cytometry, respectively. Amongst the investigated siRNAs and transfection reagents, Mcl-1/Prime-Fect complexes showed the highest inhibition of cell viability and the most effective siRNA delivery. The effect of various formulations on transfection efficiency showed that the additive with 1:1 ratio with siRNA was optimal achieving the lowest cell viability compared to untreated cells and negative control siRNA treatment (p < 0.05). Furthermore, the combination of Mcl-1 and survivin siRNA suppressed the growth of MDA-MB-436 cells more effectively than treatment with the single siRNAs and resulted in cell viability as low as ~ 20% (vs. non-treated cells). This aligned well with the induction of apoptosis as analyzed by flow cytometry, which revealed higher apoptotic cells with the combination treatment group. We conclude that commercial transfection reagents formulated with Mcl-1/Survivin siRNA combination could serve as a potent anti-proliferation agent in the treatment of breast cancers.

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Sung H, Ferlay J, Siegel RL et al (2021) Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin 71:209–249. https://doi.org/10.3322/caac.21660CrossRefPubMed

Cronin PA, Gemignani ML (2018) 14 - Breast Diseases. In: DiSaia PJ, Creasman WT, Mannel RS, McMeekin DS, Mutch DG (eds) Clinical Gynecologic Oncology (Ninth Edition). Elsevier, pp 320-352.e326CrossRef

Waks AG, Winer EP (2019) Breast Cancer Treatment: A Review. JAMA 321:288–300. https://doi.org/10.1001/jama.2018.19323CrossRefPubMed

Ngamcherdtrakul W, Yantasee W (2019) siRNA therapeutics for breast cancer: recent efforts in targeting metastasis, drug resistance, and immune evasion. Transl Res 214:105–120. https://doi.org/10.1016/j.trsl.2019.08.005CrossRefPubMedPubMedCentral

Hu B, Zhong L, Weng Y et al (2020) Therapeutic siRNA: state of the art. Signal Transduct Target Ther 5:101. https://doi.org/10.1038/s41392-020-0207-xCrossRefPubMedPubMedCentral

Santhekadur PK, Kumar DP (2020) RISC assembly and post-transcriptional gene regulation in Hepatocellular Carcinoma. Genes Dis 7:199–204. https://doi.org/10.1016/j.gendis.2019.09.009CrossRefPubMed

Wang H, Zhang S, Lv J et al (2021) Design of polymers for siRNA delivery: Recent progress and challenges. VIEW 2:20200026. https://doi.org/10.1002/VIW.20200026CrossRef

Mohseni M, Kucharski C, KC RB (2021) Therapeutic delivery of siRNA with polymeric carriers to down-regulate STAT5A expression in high-risk B-cell acute lymphoblastic leukemia (B-ALL). PLoS ONE 16:e0251719. https://doi.org/10.1371/journal.pone.0251719CrossRefPubMedPubMedCentral

Zakeri A, MaJ K, Beheshtkhoo N et al (2018) Polyethylenimine-based nanocarriers in co-delivery of drug and gene: a developing horizon. Nano Rev Exp 9:1488497CrossRef

10.

Bahadur KR, Uludağ H (2016) PEI and its derivatives for gene therapy. In: Polymers and nanomaterials for gene therapy. Elsevier, p 29–54

11.

Kurosaki T, Kitahara T, Fumoto S et al (2009) Ternary complexes of pDNA, polyethylenimine, and gamma-polyglutamic acid for gene delivery systems. Biomaterials 30:2846–2853. https://doi.org/10.1016/j.biomaterials.2009.01.055CrossRefPubMed

12.

Roesler S, Koch FP, Schmehl T et al (2011) Amphiphilic, low molecular weight poly(ethylene imine) derivatives with enhanced stability for efficient pulmonary gene delivery. J Gene Med 13:123–133. https://doi.org/10.1002/jgm.1538CrossRefPubMed

13.

Serrano-Sevilla I, Artiga Á, Mitchell SG et al (2019) Natural Polysaccharides for siRNA Delivery: Nanocarriers Based on Chitosan, Hyaluronic Acid, and Their Derivatives. Molecules 24:2570CrossRef

14.

Liao Z-X, Ho Y-C, Chen H-L et al (2010) Enhancement of efficiencies of the cellular uptake and gene silencing of chitosan/siRNA complexes via the inclusion of a negatively charged poly(γ-glutamic acid). Biomaterials 31:8780–8788. https://doi.org/10.1016/j.biomaterials.2010.07.086CrossRefPubMed

15.

Parmar MB, KC RB, Lobenberg R (2018) Additive Polyplexes to Undertake siRNA Therapy against CDC20 and Survivin in Breast Cancer Cells. Biomacromol 19:4193–4206. https://doi.org/10.1021/acs.biomac.8b00918CrossRef

16.

Zhu M, Zhang Y-M (2017) Function of myeloid cell leukaemia-1 and its regulative relations with hepatocellular carcinoma. Hepatoma Res 3:129–140. https://doi.org/10.20517/2394-5079.2017.14

17.

Campbell KJ, Dhayade S, Ferrari N et al (2018) MCL-1 is a prognostic indicator and drug target in breast cancer. Cell Death and Dis 9:19. https://doi.org/10.1038/s41419-017-0035-2CrossRef

18.

Garg SM, Falamarzian A, Vakili MR et al (2016) Polymeric micelles for MCL-1 gene silencing in breast tumors following systemic administration. Nanomedicine (Lond) 11:2319–2339. https://doi.org/10.2217/nnm-2016-0178CrossRef

19.

Thapa B, Remant KC, Uludağ H (2019) siRNA Library Screening to Identify Complementary Therapeutic Pairs in Triple-Negative Breast Cancer Cells. Methods Mol Biol 1974:1–19. https://doi.org/10.1007/978-1-4939-9220-1_1CrossRefPubMed

20.

Plianwong S, Thapa B, KC RB et al (2020) Enabling Combinatorial siRNA Delivery against Apoptosis-Related Proteins with Linoleic Acid and α-Linoleic Acid Substituted Low Molecular Weight Polyethylenimines. Pharm Res 37:46. https://doi.org/10.1007/s11095-020-2770-9CrossRefPubMed

21.

Yao T, Asayama Y (2017) Animal-cell culture media: History, characteristics, and current issues. Reprod Med Biol 16:99–117. https://doi.org/10.1002/rmb2.12024CrossRefPubMedPubMedCentral

22.

Hickerson RP, Vlassov AV, Wang Q et al (2008) Stability study of unmodified siRNA and relevance to clinical use. Oligonucleotides 18:345–354. https://doi.org/10.1089/oli.2008.0149CrossRefPubMedPubMedCentral

23.

Parmar MB, Sundaram DN, KC RB (2018) Combinational siRNA delivery using hyaluronic acid modified amphiphilic polyplexes against cell cycle and phosphatase proteins to inhibit growth and migration of triple-negative breast cancer cells. Acta Biomater 66:294–309. https://doi.org/10.1016/j.actbio.2017.11.036CrossRefPubMed

24.

Rio DC, Ares M Jr, Hannon GJ et al (2010) Purification of RNA using TRIzol (TRI reagent). Cold Spring Harb Protoc 2010:pdb.prot5439. https://doi.org/10.1101/pdb.prot5439

25.

Parmar MB, Aliabadi HM, Mahdipoor P et al (2015) Targeting Cell Cycle Proteins in Breast Cancer Cells with siRNA by Using Lipid-Substituted Polyethylenimines. Front Bioeng Biotechnol 3. https://doi.org/10.3389/fbioe.2015.00014

26.

Slastnikova TA, Ulasov AV, Rosenkranz AA et al (2018) Targeted Intracellular Delivery of Antibodies: The State of the Art. Front Pharmacol 9. https://doi.org/10.3389/fphar.2018.01208

27.

Song H, Hart SL, Du Z (2021) Assembly strategy of liposome and polymer systems for siRNA delivery. Int J Pharm 592:120033. https://doi.org/10.1016/j.ijpharm.2020.120033CrossRefPubMed

28.

Qin J-J, Yan L, Zhang J et al (2019) STAT3 as a potential therapeutic target in triple negative breast cancer: a systematic review. J Exp Clin Cancer Res 38:195. https://doi.org/10.1186/s13046-019-1206-zCrossRefPubMedPubMedCentral

29.

Zhou Z, Zhang M, Liu Y et al (2018) Reversible Covalent Cross-Linked Polycations with Enhanced Stability and ATP-Responsive Behavior for Improved siRNA Delivery. Biomacromol 19:3776–3787. https://doi.org/10.1021/acs.biomac.8b00922CrossRef

30.

Li H, Liu L, Chang H et al (2018) Downregulation of MCL-1 and upregulation of PUMA using mTOR inhibitors enhance antitumor efficacy of BH3 mimetics in triple-negative breast cancer. Cell Death and Dis 9:137. https://doi.org/10.1038/s41419-017-0169-2CrossRef

31.

Prabhakar N, Zhang J, Desai D et al (2016) Stimuli-responsive hybrid nanocarriers developed by controllable integration of hyperbranched PEI with mesoporous silica nanoparticles for sustained intracellular siRNA delivery. Int J Nanomedicine 11:6591–6608. https://doi.org/10.2147/ijn.S120611CrossRefPubMedPubMedCentral

32.

Karra H, Repo H, Ahonen I et al (2014) Cdc20 and securin overexpression predict short-term breast cancer survival. Br J Cancer 110:2905–2913CrossRef

33.

Halacli SO, Halacli B, Altundag K (2013) The significance of heat shock proteins in breast cancer therapy. Med Oncol 30:575CrossRef

34.

Curtis NL, Ruda GF, Brennan P et al (2020) Deregulation of Chromosome Segregation and Cancer. Annu Rev Cancer Biol 4:257–278. https://doi.org/10.1146/annurev-cancerbio-030419-033541CrossRef

35.

Ali MMU, Roe SM, Vaughan CK et al (2006) Crystal structure of an Hsp90–nucleotide–p23/Sba1 closed chaperone complex. Nature 440:1013–1017. https://doi.org/10.1038/nature04716CrossRefPubMedPubMedCentral

36.

Michels J, Johnson PWM, Packham G (2005) Mcl-1. Int J Biochem Cell Biol 37:267–271. https://doi.org/10.1016/j.biocel.2004.04.007CrossRefPubMed

37.

Wang T, Larcher LM, Ma L et al (2018) Systematic Screening of Commonly Used Commercial Transfection Reagents towards Efficient Transfection of Single-Stranded Oligonucleotides. Molecules (Basel, Switzerland) 23:2564. https://doi.org/10.3390/molecules23102564CrossRef

38.

Zaffaroni N, Daidone MG (2002) Survivin expression and resistance to anticancer treatments: perspectives for new therapeutic interventions. Drug Resist Updat 5:65–72. https://doi.org/10.1016/s1368-7646(02)00049-3CrossRefPubMed

39.

Jha K, Shukla M, Pandey M (2012) Survivin expression and targeting in breast cancer. Surg Oncol 21:125–131. https://doi.org/10.1016/j.suronc.2011.01.001CrossRefPubMed

40.

Norouzi P, Motasadizadeh H, Atyabi F et al (2021) Combination Therapy of Breast Cancer by Codelivery of Doxorubicin and Survivin siRNA Using Polyethylenimine Modified Silk Fibroin Nanoparticles. ACS Biomater Sci Eng 7:1074–1087. https://doi.org/10.1021/acsbiomaterials.0c01511CrossRefPubMed

41.

Pengnam S, Plianwong S, Patrojanasophon P et al (2021) Synergistic Effect of Doxorubicin and siRNA-Mediated Silencing of Mcl-1 Using Cationic Niosomes against 3D MCF-7 Spheroids. Pharmaceutics 13. https://doi.org/10.3390/pharmaceutics13040550

42.

Thapa B, Kc R, Liu X et al (2022) TRAIL Therapy for Breast Cancer Treatment by Employing Lipopolymer mRNA Delivery. GEN Biotechnology 1:101–112. https://doi.org/10.1089/genbio.2021.0007CrossRef

43.

Aliabadi HM, Maranchuk R, Kucharski C et al (2013) Effective response of doxorubicin-sensitive and -resistant breast cancer cells to combinational siRNA therapy. J Controll Release 172:219–228. https://doi.org/10.1016/j.jconrel.2013.08.012CrossRef

44.

Ueda A, Oikawa K, Fujita K et al (2019) Therapeutic potential of PLK1 inhibition in triple-negative breast cancer. Lab Invest 99:1275–1286. https://doi.org/10.1038/s41374-019-0247-4CrossRefPubMed

45.

Domínguez-Gómez G, Díaz-Chávez J, Chávez-Blanco A et al (2015) Nicotinamide sensitizes human breast cancer cells to the cytotoxic effects of radiation and cisplatin. Oncol Rep 33:721–728. https://doi.org/10.3892/or.2014.3661CrossRefPubMed

46.

Zong B, Sun L, Peng Y et al (2021) HORMAD1 promotes docetaxel resistance in triple negative breast cancer by enhancing DNA damage tolerance. Oncol Rep 46. https://doi.org/10.3892/or.2021.8089

47.

Janghorban M, Farrell AS, Allen-Petersen BL et al (2014) Targeting c-MYC by antagonizing PP2A inhibitors in breast cancer. Proc Natl Acad Sci USA 111:9157–9162. https://doi.org/10.1073/pnas.1317630111CrossRefPubMedPubMedCentral

Titel: Improved delivery of Mcl-1 and survivin siRNA combination in breast cancer cells with additive siRNA complexes
verfasst von: Tinnabhop Santadkha
Wanwisa Skolpap
Remant K.C.
Aysha Ansari
Cezary Kucharski
Teo Atz Dick
Hasan Uludağ
Publikationsdatum: 14.07.2022
Verlag: Springer US
Erschienen in: Investigational New Drugs / Ausgabe 5/2022
Print ISSN: 0167-6997
Elektronische ISSN: 1573-0646
DOI: https://doi.org/10.1007/s10637-022-01282-y

Springer Medizin

Improved delivery of Mcl-1 and survivin siRNA combination in breast cancer cells with additive siRNA complexes

Summary

Neu im Fachgebiet Onkologie

Labor, CT-Anthropometrie zeigen Risiko für Pankreaskrebs

Viel pflanzliche Nahrung, seltener Prostata-Ca.-Progression

Alter verschlechtert Prognose bei Endometriumkarzinom

Darf man die Behandlung eines Neonazis ablehnen?

Update Onkologie

Springer Medizin

Summary

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