Downregulation of LPXN expression by siRNA decreases the malignant proliferation and transmembrane invasion of SHI‑1 cells

Zhu,Guo‑Hua; Dai,Hai‑Ping; Shen,Qun; Zhang,Qi

doi:10.3892/ol.2018.9605

Downregulation of LPXN expression by siRNA decreases the malignant proliferation and transmembrane invasion of SHI‑1 cells

Authors:
- Guo‑Hua Zhu
- Hai‑Ping Dai
- Qun Shen
- Qi Zhang
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Affiliations: First Clinical College, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, P.R. China, Leukemia Research Unit, Jiangsu Institute of Hematology, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China
Published online on: October 22, 2018 https://doi.org/10.3892/ol.2018.9605
Pages: 135-140
Copyright: © Zhu 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 investigate the effects of decreasing leupaxin (LPXN) expression on the proliferation and invasion of human acute monocytic leukemia SHI‑1 cells. The transfection efficiency of fluorescein amidite (FAM)‑small interfering RNA (siRNA) was determined using flow cytometry, and the protein expression levels of LPXN, phosphorylated (p)‑c‑Jun N‑terminal kinase (JNK), p‑p38 mitogen‑activated protein kinase (p38 MAPK) and p‑extracellular‑signal‑regulated kinase (ERK) were detected by western blot analysis. Proliferation was determined using the cell counting kit‑8 reagent and cellular transmembrane invasion ability was determined using a Transwell chamber system. The gelatinase levels of matrix metalloproteinase (MMP)‑2 and MMP‑9 in the cell culture supernatant were also analyzed by gelatin zymography. In SHI‑1 cells, the optimal transfection conditions of siRNA were a cell density of 4x105 cells/ml and a ratio of siRNA/Lipofectamine® 2000 of 200 pmol/1 µl. The highest transfection efficiency of FAM‑siRNA was 74.5%. In the present study, L2‑siRNA was selected to effectively decrease the expression of LPXN. Following downregulation of LPXN expression by L2‑siRNA, proliferation inhibition rates increased to 27.043±2.051 and cell transmembrane invasion rates decreased to 25.270±2.145 (P<0.05). The results of the western blot analysis and the gelatin zymography indicated that downregulation of LPXN expression increased the expression of p‑p38 MAPK and p‑JNK, and attenuated the secretion levels of MMP‑2 and MMP‑9. However, downregulation of LPXN expression had no effect on p‑ERK expression in SHI‑1 cells. The results of the present study indicated that downregulation of LPXN expression decreased the malignant proliferation and transmembrane invasion of SHI‑1 cells by activating JNK and p38 MAPK, and inhibiting MMP‑2 and MMP‑9 secretion.

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1	Lipsky BP, Beals CR and Staunton DE: Leupaxin is a novel LIM domain protein that forms a complex with PYK2. J Biol Chem. 273:11709–22713. 1998. View Article : Google Scholar : PubMed/NCBI
2	Chen PW and Kroog GS: Leupaxin is similar to paxillin in focal adhesion targeting and tyrosine phosphorylation but has distinct roles in cell adhesion and spreading. Cell Adh Migr. 4:527–540. 2010. View Article : Google Scholar : PubMed/NCBI
3	Deakin NO, Pignatelli J and Turner CE: Diverse roles for the paxillin family of proteins in cancer. Genes Cancer. 3:362–370. 2015. View Article : Google Scholar
4	Kaulfuss S, von Hardenberg S, Schweyer S, Herr AM, Laccone F, Wolf S and Burfeind P: Leupaxin acts as a mediator in prostate carcinoma progression through deregulation of p120catenin expression. Oncogene. 28:3971–3982. 2009. View Article : Google Scholar : PubMed/NCBI
5	Watanabe N, Amano N, Ishizuka H and Mashima K: Leupaxin binds to PEST domain tyrosine phosphatase PEP. Mol Cell Biochem. 269:13–17. 2005. View Article : Google Scholar : PubMed/NCBI
6	Sundberg-Smith LJ, DiMichele LA, Sayers RL, Mack CP and Taylor JM: The LIM protein leupaxin is enriched in smooth muscle and functions as an serum response factor cofactor to induce smooth muscle cell gene transcription. Circ Res. 102:1502–1511. 2008. View Article : Google Scholar : PubMed/NCBI
7	Petti F, Thelemann A, Kahler J, McCormack S, Castaldo L, Hunt T, Nuwaysir L, Zeiske L, Haack H, Sullivan L, et al: Temporal quantitation of mutant kit tyrosine kinase signaling attenuated by a novel thiophene kinase inhibitor OSI-930. Mol Cancer Ther. 4:1186–1197. 2005. View Article : Google Scholar : PubMed/NCBI
8	Tanaka T, Moriwaki K, Murata S and Miyasaka M: LIM domain-containing adaptor, leupaxin, localizes in focal adhesion and suppresses the integrin-induced tyrosine phosphorylation of paxillin. Cancer Sci. 101:363–368. 2010. View Article : Google Scholar : PubMed/NCBI
9	Dai HP, Xue YQ, Zhou JW, Li AP, Wu YF, Pan JL, Wang Y and Zhang J: LPXN, a member of the paxillin superfamily, is fused to RUNX1 in an acute myeloid leukemia patient with a t(11;21)(q12;q22) translocation. Genes Chromosomes Cancer. 48:1027–1036. 2009. View Article : Google Scholar : PubMed/NCBI
10	Abe A, Yamamoto Y, Iba S, Kanie T, Okamoto A, Tokuda M, Inaguma Y, Yanada M, Morishima S, Mizuta S, et al: ETV6-LPXN fusion transcript generated by t(11;12) (q12.1;p13) in a patient with relapsing acute myeloid leukemia with NUP98-HOXA9. Genes Chromosomes Cancer. 55:242–250. 2016. View Article : Google Scholar : PubMed/NCBI
11	Wang CL, Chen ZX, Li ZJ and Cen J: The essential roles of matrix metalloproteinase-2, membrane type 1 metalloproteinase and tissue inhibitor of metalloproteinase-2 in the invasive capacity of acute monocytic leukemia SHI-1 cells. Leuk Res. 34:1083–1090. 2010. View Article : Google Scholar : PubMed/NCBI
12	Kaulfuss S, Grzmil M, Hemmerlein B, Thelen P, Schweyer S, Neesen J, Bubendorf L, Glass AG, Jarry H, Auber B, et al: Leupaxin, a novel coactivator of the androgen receptor, is expressed in prostate cancer and plays a role in adhesion and invasion of prostate carcinoma cells. Mol Endocrinol. 22:1606–1621. 2008. View Article : Google Scholar : PubMed/NCBI
13	Zhu GH, Dai HP, Shen Q, Ji O, Zhang Q and Zhai YL: Curcumin induces apoptosis and suppresses invasion through MAPK and MMP signaling in human monocytic leukemia SHI-1 cells. Pharm Biol. 54:1303–1311. 2016.PubMed/NCBI
14	Schaller MD: Paxillin: A focal adhesion-associated adapter protein. Oncogene. 20:6459–6472. 2001. View Article : Google Scholar : PubMed/NCBI
15	van Nimwegen MJ and van de Water B: Focal adhesion kinase: A potential target in cancer therapy. Biochem Pharmacol. 73:597–609. 2007. View Article : Google Scholar : PubMed/NCBI
16	Calderwood DA, Shattil SJ and Ginsberg MH: Integrins and actin filaments: Reciprocal regulation of cell adhesion and signaling. J Biol Chem. 275:22607–22610. 2000. View Article : Google Scholar : PubMed/NCBI
17	Omari S: Focal adhesion kinase: A key mediator of cancer pathogenesis. HealthMED. 5:807–818. 2011.
18	Sahu SN, Nunez S, Bai G and Gupta A: Interaction of Pyk2 and PTP-PEST with leupaxin in prostate cancer cells. Am J Physiol Cell Physiol. 292:C2288–C2296. 2007. View Article : Google Scholar : PubMed/NCBI
19	Shi J, Wu WJ, Hu G, Yu X, Yu GS, Lu H, Yang ML, Liu B and Wu ZX: Regulation of β-catenin transcription activity by leupaxin in hepatocellular carcinoma. Tumour Biol. 37:2313–2320. 2016. View Article : Google Scholar : PubMed/NCBI
20	Sahu SN, Khadeer MA, Robertson BW, Núñez SM, Bai G and Gupta A: Association of leupaxin with Src in osteoclasts. Am J Physiol Cell Physiol. 292:C581–C590. 2007. View Article : Google Scholar : PubMed/NCBI
21	Kaulfuss S, Herr AM, Büchner A, Hemmerlein B, Günthert AR and Burfeind P: Leupaxin is expressed in mammary carcinoma and acts as a transcriptional activator of the estrogen receptor α. Int J Oncol. 47:106–114. 2015. View Article : Google Scholar : PubMed/NCBI
22	Vanarotti MS, Finkelstein DB, Guibao CD, Nourse A, Miller DJ and Zheng JJ: Structural basis for the interaction between Pyk2-FAT domain and leupaxin LD repeats. Biochemistry. 55:1332–1345. 2016. View Article : Google Scholar : PubMed/NCBI
23	Dierks S, von Hardenberg S, Schmidt T, Bremmer F, Burfeind P and Kaulfuss S: Leupaxin stimulates adhesion and migration of prostate cancer cells through modulation of the phosphorylation status of the actin-binding protein caldesmon. Oncotarget. 6:13591–13606. 2015. View Article : Google Scholar : PubMed/NCBI
24	Chen DL, Wang ZQ, Ren C, Zeng ZL, Wang DS, Luo HY, Wang F, Qiu MZ, Bai L, Zhang DS, et al: Abnormal expression of paxillin correlates with tumor progression and poor survival in patients with gastric cancer. J Transl Med. 11:2772013. View Article : Google Scholar : PubMed/NCBI
25	Chew V and Lam KP: Leupaxin negatively regulates B cell receptor signaling. J Biol Chem. 282:27181–27191. 2007. View Article : Google Scholar : PubMed/NCBI