Different responses of cell cycle between rat vascular smooth muscle cells and vascular endothelial cells to paclitaxel

Abstract

Although previous reports showed drug-eluting stent (DES) could effectively inhibit neointima formation, in-stent restenosis (ISR) remains an important obstacle. The purpose of this study was to investigate different effects of paclitaxel on proliferation and cell cycle regulators between vascular smooth muscle cells (VSMCs) and vascular endothelial cells (VECs) of rats in vitro. The cultured VSMCs and VECs of rats from the same tissues were examined by using immunohistochemistry, flow cytometry and Western blotting in control and paclitaxel-treated groups. The results showed paclitaxel could effectively inhibit proliferation of VSMCs and VECs. However, as compared with VECs, proliferation of VSMCs in paclitaxel-treated group decreased less rapidly. The percentage of cells in G₀-G₁ and G₂-M phases was reduced, and that in S phase increased after treatment for 72 h. The expression of cyclin D1 and B1, p27 and PCNA in VSMCs of paclitaxel-treated group was up-regulated, but that of p21 down-regulated as compared with VECs. It is concluded that there are significant differences in the expression of cell cycle regulators and proliferation rate between paclitaxel-treated VSMCs and paclitaxel-treated VECs, suggesting that the G₁-S checkpoint regulated by paclitaxel may play a critical role in the development of complications of DES, which provides new strategies for treatments of ISR.

Keywords

vascular smooth muscle cells / vascular endothelial cells / paclitaxel / drug-eluting stent

References

[1] LohJP, WaksmanR. Paclitaxel drug-coated balloons: a review of current status and emerging applications in native coronary artery de novo lesions. JACC Cardiovasc Interv, 2012, 5(10): 1001-1012

[2] KoYG, KimJS, KimBK, et al.. Efficacy of drug-eluting stents for treating in-stent restenosis of drug-eluting stents (from the Korean DES ISR multicenter registry study [KISS]). Am J Cardiol, 2012, 109(5): 607-613

[3] KibosA, CampeanuA, TintoiuI, et al.. Pathophysiology of coronary artery in-stent restenosis. Acute Card Care, 2007, 9(2): 111-119

[4] QiuJ, GaoHQ, LiBY, et al.. Proteomics investigation of protein expression changes in ouabain induced apoptosis in human umbilical vein endothelial cells. J Cell Biochem, 2008, 104(3): 1054-1064

[5] InoueT, NodeK. Molecular basis of restenosis and novel issues of drug-eluting stents. Circ J, 2009, 73(4): 615-621

[6] Ribamar CostaJ, SousaAG, MoreiraA, et al.. Comparison of the very long term (>1 year) outcomes of drug-eluting stents for the treatment of bare-metal and drug-eluting stent restenosis. Euro Intervention, 2009, 5(4): 448-453

[7] LeeSE, JohnSH, LimJH, et al.. Very late stent thrombosis associated with multiple stent fractures and peri-stent aneurysm formation after sirolimus-eluting stent implantation. Circ J, 2008, 72(7): 1201-1204

[8] LukSC, SiuSW, LaiCK, et al.. Cell cycle arrest by a natural product via G2/M checkpoint. Int J Med Sci, 2005, 5(5): 64-69

[9] BoyeE, SkjolbergHC, GrallertB. Checkpoint regulation of DNA replication. Methods Mol Biol, 2009, 521: 55-70

[10] CollinsI, GarrettMD. Targeting the cell division cycle in cancer: CDK and cell cycle checkpoint kinase inhibitors. Curr Opin Pharmacol, 2005, 5(4): 366-373

[11] KobayashiM, InoueK, WarabiE, et al.. A simple method of isolating mouse aortic endothelial cells. J Atheroscler Thromb, 2005, 12(3): 138-142

[12] WangDG, LiuZY, BiYS, et al.. Culture of smooth muscle cells from guinea pig mesenteric lymphatic vessels. Lymphology, 2007, 40(1): 14-18

[13] MunariLM, PortaM. Cerebrovascular disease: pathophysiology and clinical aspects. Rays, 1990, 15(2): 153-164

[14] BreenDM, ChanKK, DhaliwallJK, et al.. Insulin increases reendothelialization and inhibits cell migration and neointimal growth after arterial injury. Arterioscler Thromb Vasc Biol, 2009, 29(7): 1060-1066

[15] SunZS, ZhouSH, GuanX. Impact of blood circulation on reendothelialization, restenosis and atrovastatin’s restenosis prevention effects. Int J Cardiol, 2008, 128(2): 261-8

[16] El-KarehAW, LabesRE, SecombTW. Cell cycle checkpoint models for cellular pharmacology of paclitaxel and platinum drugs. AAPS J, 2008, 10(1): 15-34

[17] SrokaIM, HeissEH, HavlicekL, et al.. A novel roscovitine derivative potently induces G1-phase arrest in platelet-derived growth factor-BB-activated vascular smooth muscle cells. Mol Pharmacol, 2010, 77(2): 255-261

[18] ThipparaboinaR, KhanW, DombAJ. Eluting combination drugs from stents. Int J Pharm, 2013

[19] MasakiT, KamerathCD, KimSJ, et al.. In vitro pharmacological inhibition of human vascular smooth muscle cell proliferation for the prevention of hemodialysis vascular access stenosis. Blood Purif, 2004, 22(3): 307-312

[20] XiaoZ, XueJ, SowinTJ, et al.. Differential roles of checkpoint kinase 1, checkpoint kinase 2, and mitogen-activated protein kinase-activated protein kinase 2 in mediating DNA damage-induced cell cycle arrest: implications for cancer therapy. Mol Cancer Ther, 2006, 5(8): 1935-1943

[21] KlopfleischR, GruberAD. Differential expression of cell cycle regulators p21, p27 and p53 in metastasizing canine mammary adenocarcinomas versus normal mammary glands. Res Vet Sci, 2009, 87(1): 91-6

[22] HiraiH, RousselMF, KatoJY, et al.. Novel INK4 proteins, p19 and p18, are specific inhibitors of the cyclin D-dependent kinases CDK4 and CDK6. Mol Cell Biol, 1995, 15(5): 2672-2681

[23] ButlerJE, SpearBT. Normal intestinal epithelial cell differentiation in the absence of p21 and p27: new insights from old knock-out mice. Cancer Biol Ther, 2008, 7(6): 880-881

[24] FascianoS, PatelRC, HandyI, et al.. Regulation of vascular smooth muscle proliferation by heparin: inhibition of cyclin-dependent kinase 2 activity by p27(kip1). J Biol Chem, 2005, 280(16): 15682-15689

[25] MasamhaCP, BenbrookDM. Cyclin D1 degradation is sufficient to induce G1 cell cycle arrest despite constitutive expression of cyclin E2 in ovarian cancer cells. Cancer Res, 2009, 69(16): 6565-6572

[26] ElmoreLW, DiX, DumurC, et al.. Evasion of a single-step, chemotherapy-induced senescence in breast cancer cells: implications for treatment response. Clin Cancer Res, 2005, 11(7): 2637-2643

[27] ZhaoYM, ZhouQ, XuY, et al.. Antiproliferative effect of rapamycin on human T-cell leukemia cell line Jurkat by cell cycle arrest and telomerase inhibition. Acta Pharmacol Sin, 2008, 29(4): 481-488

[28] TirochK, KochW, MehilliJ, et al.. P27 and P53 gene polymorphisms and restenosis following coronary implantation of drug-eluting stents. Cardiology, 2009, 112(4): 263-269

[29] FuchsAT, KuehnlA, PelisekJ, et al.. Inhibition of restenosis formation without compromising reendothelialization as a potential solution to thrombosis following angioplasty?. Endothelium, 2008, 15(1): 85-92