Background: Arterial wall is exposed to three main hemodynamic forces in vivo: shear stress, caused by the dragging frictional force of blood flow; transmural pressure, caused by the hydrostatic forces of blood within the blood vessel; and cyclic strain, defined as the repetitive deformation of the cells as the arterial wall rhythmically distends and relaxes with the cardiac cycle [1]. It had been proved that pathophysiological mechanical states of vascular cells, i.e. endothelial cells (ECs) and vascular smooth muscle cells (VSMCs), contribute to vascular disorder resulting from atherosclerosis, hypertension, bypass graft occlusion, et al [1, 2]. Nucleus is the largest and stiffest organelle in the cell, where is the site of transcriptional regulation, and contains the genome [3]. Nuclear matrix acts as the skeleton structure of the nucleus, but its role in mechano-sensing and gene-expression-regulating is still unclear. Methods and Results: The co-culture parallel-plate flow chamber system was used to mimics the shear stress, and FX-4000T Strain Unit to mimics the cyclic strain, respectively. We demonstrated the effects of three kinds of nuclear matrix proteins, i.e. Nesprin2, SUN1 and LaminA, in proliferation and apoptosis of vascular cells modulated by shear stress and cyclic strain. Our results indicated that both shear stress and cyclic strain application significantly modulated the expressions of Nesprin2, SUN1 and LaminA, but in different ways. Physiological shear stress (15 dyne/cm2) and pathological shear stress (5 dyne/cm2) revealed similar repressed effect on expressions of nuclear matrix in comparison with the static control. Physiological cyclic strain (5% elongation) increased, but pathological (15% elongation) decreased the expressions of nuclear matix proteins. By using target siRNA and overexpression plasmid transfection in ECs and VSMCs respectively, it revealed that nuclear matrix proteins participated in regulation of apoptosis and proliferation. Furthermore, the activations of transcription factors were detected by Protein/DNA array after both suppressing and increasing nuclear matrix proteins expressions. TFIID, regulated by Nesprin2, Stat-1, 3, 5 and 6, regulated by LaminA, were remarkably changed in vascular cells transfected with target siRNA and overexpression plasmid respectively. Conclusion: Our study revealed that nuclear matrix proteins might be the novel mechanical-sensitive molecules in vascular cells, which and may play a crucial role in cell functions.