MicroRNA‑223 overexpression suppresses protein kinase C &epsilon; expression in human leukemia stem cell‑like KG‑1a cells

Osiriphan,Mallika; Insukhin,Charapat; Anuchapreeda,Songyot; Khamphikham,Pinyaphat; Duangmano,Suwit

doi:10.3892/mco.2024.2746

July-2024 Volume 21 Issue 1

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July-2024 Volume 21 Issue 1

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Article Open Access

MicroRNA‑223 overexpression suppresses protein kinase C ε expression in human leukemia stem cell‑like KG‑1a cells

Authors:
- Mallika Osiriphan
- Charapat Insukhin
- Songyot Anuchapreeda
- Pinyaphat Khamphikham
- Suwit Duangmano
View Affiliations / Copyright

Affiliations: Division of Clinical Microscopy, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Mueang Chiang Mai, Chiang Mai 50200, Thailand

Copyright: © Osiriphan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Article Number: 48
|
Published online on: May 28, 2024

https://doi.org/10.3892/mco.2024.2746
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Abstract

MicroRNA‑223 (miR‑223) is dysregulated in various cancer types, including acute myeloid leukemia (AML). Despite this, there has been a lack of studies exploring the role of miR‑223 in leukemic stem cells, particularly those involved in drug resistance, a major cause of chemotherapy failure in AML. The present study aimed to elucidate the impact of miR‑223 on drug resistance in the leukemic stem‑cell line, KG‑1a. Two AML cell lines, KG‑1 and KG‑1a, differing in the proportion of CD34⁺CD38^‑ cells, were assessed for doxorubicin (DOX) sensitivity using the Cell Counting Kit‑8 assay. The expression levels of miR‑223 and protein kinase C ε (PKCε) were evaluated via reverse transcription‑quantitative PCR and western blot analysis. The association between miR‑223 and its target, PKCε, was confirmed by luciferase activity assay. The effects of miR‑223 overexpression and PKCε inhibition were also evaluated in KG‑1a cells using miR‑223 mimic and small interfering (si)RNA transfection, respectively. Daunorubicin was then used to assess drug sensitivity in the siRNA‑transfected KG‑1a cells. Compared with KG‑1 cells, KG‑1a cells displayed greater resistance to DOX, and had increased PKCε levels and decreased miR‑223 expression. Overexpression of miR‑223 led to PKCε protein downregulation in KG‑1a cells, which was further confirmed by a luciferase assay demonstrating miR‑223 targeting of PKCε. However, despite these effects, miR‑223 overexpression and PKCε inhibition did not change drug sensitivity in KG‑1a cells compared with negative control cells. In summary, the present study demonstrated that miR‑223 could target and silence PKCε expression in KG‑1a cells; however, the chemoresistance of KG‑1a cells to anthracycline drugs may not be directly associated with the low expression of miR‑223.

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1	Deschler B and Lübbert M: Acute myeloid leukemia: Epidemiology and etiology. Cancer. 107:2099–2107. 2006.PubMed/NCBI View Article : Google Scholar
2	Pelcovits A and Niroula R: Acute myeloid leukemia: A review. R I Med J (2013). 103:38–40. 2020.PubMed/NCBI
3	Vakiti A and Mewawalla P: Acute myeloid leukemia. In: StatPearls [Internet], StatPearls Publishing, Treasure Island, FL, 2022.
4	Zhang J, Gu Y and Chen B: Mechanisms of drug resistance in acute myeloid leukemia. Onco Targets Ther. 12:1937–1945. 2019.PubMed/NCBI View Article : Google Scholar
5	Marchand T and Pinho S: Leukemic stem cells: From leukemic niche biology to treatment opportunities. Front Immunol. 12(775128)2021.PubMed/NCBI View Article : Google Scholar
6	Hanekamp D, Cloos J and Schuurhuis GJ: Leukemic stem cells: Identification and clinical application. Int J Hematol. 105:549–557. 2017.PubMed/NCBI View Article : Google Scholar
7	Jordan CT: The leukemic stem cell. Best Pract Res Clin Haematol. 20:13–18. 2007.PubMed/NCBI View Article : Google Scholar
8	Wang X, Huang S and Chen JL: Understanding of leukemic stem cells and their clinical implications. Mol Cancer. 16(2)2017.PubMed/NCBI View Article : Google Scholar
9	Phi LTH, Sari IN, Yang YG, Lee SH, Jun N, Kim KS, Lee YK and Kwon HY: Cancer stem cells (CSCs) in drug resistance and their therapeutic implications in cancer treatment. Stem Cells Int. 2018(5416923)2018.PubMed/NCBI View Article : Google Scholar
10	Aziz F, Chakraborty A, Khan I and Monts J: Relevance of miR-223 as potential diagnostic and prognostic markers in cancer. Biology (Basel). 11(249)2022.PubMed/NCBI View Article : Google Scholar
11	Gentner B, Pochert N, Rouhi A, Boccalatte F, Plati T, Berg T, Sun SM, Mah SM, Mirkovic-Hösle M, Ruschmann J, et al: MicroRNA-223 dose levels fine tune proliferation and differentiation in human cord blood progenitors and acute myeloid leukemia. Exp Hematol. 43:858–868.e7. 2015.PubMed/NCBI View Article : Google Scholar
12	Yu G, Yin Z, He H, Zheng Z, Chai Y, Xuan L, Lin R, Wang Q, Li J and Xu D: Low serum miR-223 expression predicts poor outcome in patients with acute myeloid leukemia. J Clin Lab Anal. 34(e23096)2020.PubMed/NCBI View Article : Google Scholar
13	Xiao Y, Su C and Deng T: miR-223 decreases cell proliferation and enhances cell apoptosis in acute myeloid leukemia via targeting FBXW7. Oncol Lett. 12:3531–3536. 2016.PubMed/NCBI View Article : Google Scholar
14	Ding L, Wang H, Lang W and Xiao L: Protein kinase C-epsilon promotes survival of lung cancer cells by suppressing apoptosis through dysregulation of the mitochondrial caspase pathway. J Biol Chem. 277:35305–35313. 2002.PubMed/NCBI View Article : Google Scholar
15	Kinehara M, Kawamura S, Tateyama D, Suga M, Matsumura H, Mimura S, Hirayama N, Hirata M, Uchio-Yamada K, Kohara A, et al: Protein kinase C regulates human pluripotent stem cell self-renewal. PLoS One. 8(e54122)2013.PubMed/NCBI View Article : Google Scholar
16	Flescher E and Rotem R: Protein kinase C epsilon mediates the induction of P-glycoprotein in LNCaP prostate carcinoma cells. Cell Signal. 14:37–43. 2002.PubMed/NCBI View Article : Google Scholar
17	Huang B, Cao K, Li X, Guo S, Mao X, Wang Z, Zhuang J, Pan J, Mo C, Chen J and Qiu S: The expression and role of protein kinase C (PKC) epsilon in clear cell renal cell carcinoma. J Exp Clin Cancer Res. 30(88)2011.PubMed/NCBI View Article : Google Scholar
18	Wang H, Zhan M, Xu SW, Chen W, Long MM, Shi YH, Liu Q, Mohan M and Wang J: miR-218-5p restores sensitivity to gemcitabine through PRKCE/MDR1 axis in gallbladder cancer. Cell Death Dis. 8(e2770)2017.PubMed/NCBI View Article : Google Scholar
19	Zhang GF, Wu JC, Wang HY, Jiang WD and Qiu L: Overexpression of microRNA-205-5p exerts suppressive effects on stem cell drug resistance in gallbladder cancer by down-regulating PRKCE. Biosci Rep. 40(BSR20194509)2020.PubMed/NCBI View Article : Google Scholar
20	Nicholson R, Menezes AC, Azevedo A, Leckenby A, Davies S, Seedhouse C, Gilkes A, Knapper S, Tonks A and Darley RL: Protein kinase C epsilon overexpression is associated with poor patient outcomes in aml and promotes daunorubicin resistance through p-glycoprotein-mediated drug efflux. Front Oncol. 12(840046)2022.PubMed/NCBI View Article : Google Scholar
21	Skopek R, Palusińska M, Kaczor-Keller K, Pingwara R, Papierniak-Wyglądała A, Schenk T, Lewicki S, Zelent A and Szymański Ł: Choosing the right cell line for acute myeloid leukemia (AML) research. Int J Mol Sci. 24(5377)2023.PubMed/NCBI View Article : Google Scholar
22	Koeffler HP, Billing R, Lusis AJ, Sparkes R and Golde DW: An undifferentiated variant derived from the human acute myelogenous leukemia cell line (KG-1). Blood. 56:265–273. 1980.PubMed/NCBI
23	Varkonyi-Gasic E and Hellens RP: Quantitative stem-loop RT-PCR for detection of microRNAs. In: RNAi and plant gene function analysis: Methods and protocols. Kodama H and Komamine A (eds). Humana Press, Totowa, NJ, pp145-157, 2011.
24	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.PubMed/NCBI View Article : Google Scholar
25	Lopes Gde L Jr, de Souza JA and Barrios C: Access to cancer medications in low- and middle-income countries. Nat Rev Clin Oncol. 10:314–322. 2013.PubMed/NCBI View Article : Google Scholar
26	Cherny N, Sullivan R, Torode J, Saar M and Eniu A: ESMO European consortium study on the availability, out-of-pocket costs and accessibility of antineoplastic medicines in Europe. Ann Oncol. 27:1423–1443. 2016.PubMed/NCBI View Article : Google Scholar
27	Jain M and Mukherjee K: Economic burden of breast cancer to the households in Punjab, India. Int J Med Public Health. 6(13)2016.
28	Ruff P, Al-Sukhun S, Blanchard C and Shulman LN: Access to cancer therapeutics in low- and middle-income countries. Am Soc Clin Oncol Educ Book. 35:58–65. 2016.PubMed/NCBI View Article : Google Scholar
29	Kaiser AH, Rotigliano N, Flessa S, Ekman B and Sundewall J: Extending universal health coverage to informal workers: A systematic review of health financing schemes in low- and middle-income countries in Southeast Asia. PLoS One. 18(e0288269)2023.PubMed/NCBI View Article : Google Scholar
30	Chueahongthong F, Tima S, Chiampanichayakul S, Berkland C and Anuchapreeda S: Co-treatments of edible curcumin from turmeric rhizomes and chemotherapeutic drugs on cytotoxicity and FLT3 protein expression in leukemic stem cells. Molecules. 26(5785)2021.PubMed/NCBI View Article : Google Scholar
31	Panyajai P, Amnajphook N, Keawsangthongcharoen S, Chiampanichayakul S, Tima S and Anuchapreeda S: Study of leukemic stem cell population (CD34+/CD38-) and WT1 protein expression in human leukemic cell lines. J Assoc Med Sci. 51:38–44. 2018.
32	Niu J, Peng D and Liu L: Drug resistance mechanisms of acute myeloid leukemia stem cells. Front Oncol. 12(896426)2022.PubMed/NCBI View Article : Google Scholar
33	Zhang N, Su Y and Xu L: Targeting PKCε by miR-143 regulates cell apoptosis in lung cancer. FEBS Lett. 587:3661–3667. 2013.PubMed/NCBI View Article : Google Scholar
34	Oliveto S, Mancino M, Manfrini N and Biffo S: Role of microRNAs in translation regulation and cancer. World J Biol Chem. 8(45)2017.PubMed/NCBI View Article : Google Scholar
35	O'Brien J, Hayder H, Zayed Y and Peng C: Overview of MicroRNA biogenesis, mechanisms of actions, and circulation. Front Endocrinol (Lausanne). 9(402)2018.PubMed/NCBI View Article : Google Scholar
36	Jin HY, Gonzalez-Martin A, Miletic AV, Lai M, Knight S, Sabouri-Ghomi M, Head SR, Macauley MS, Rickert RC and Xiao C: Transfection of microRNA mimics should be used with caution. Front Genet. 6(340)2015.PubMed/NCBI View Article : Google Scholar
37	Inoue K, Jinnin M, Yamane K, Makino T, Kajihara I, Makino K, Honda N, Nakayama W, Fukushima S and Ihn H: Down-regulation of miR-223 contributes to the formation of Gottron's papules in dermatomyositis via the induction of PKCε. Eur J Dermatol. 23:160–167. 2013.PubMed/NCBI View Article : Google Scholar
38	Khan K, Zafar S, Badshah Y, Ashraf NM, Rafiq M, Danish L, Shabbir M, Trembley JH, Afsar T, Almajwal A and Razak S: Cross talk of tumor protein D52 (TPD52) with KLF9, PKCε, and MicroRNA 223 in ovarian cancer. J Ovarian Res. 16(202)2023.PubMed/NCBI View Article : Google Scholar
39	Ding J, Zhao Z, Song J, Luo B and Huang L: MiR-223 promotes the doxorubicin resistance of colorectal cancer cells via regulating epithelial-mesenchymal transition by targeting FBXW7. Acta Biochim Biophys Sin (Shanghai). 50:597–604. 2018.PubMed/NCBI View Article : Google Scholar
40	Gobbi G, Di Marcantonio D, Micheloni C, Carubbi C, Galli D, Vaccarezza M, Bucci G, Vitale M and Mirandola P: TRAIL up-regulation must be accompanied by a reciprocal PKCε down-regulation during differentiation of colonic epithelial cell: Implications for colorectal cancer cell differentiation. J Cell Physiol. 227:630–638. 2012.PubMed/NCBI View Article : Google Scholar
41	Nicholson RL, Knapper S, Tonks A and Darley RL: PKC-Epsilon overexpression is associated with poor outcomes in AML and promotes chemoresistance and hematopoietic stem cell quiescence. Blood. 134 (Suppl 1)(S2704)2019.