Down-regulation of Notch1 inhibits proliferation and induces apoptosis of fibroblasts from pathological scars

Abstract

Abstract: Objective To investigate the effect of down-regulation of Notch1 expression mediated by lentivirus on the activation of caspase-9 and mitochondrial membrane potential of fibroblast from pathological scars(PSF). Methods PSFs were isolated and cultured and then infected by Notch1 siRNA lentivirus and negative control lentivirus respectively.The level of Notch1 mRNA and protein was detected by real-time PCR and Western blot,respectively.MTT assay was used to detect cell proliferation; PI monochrome staining was used to detect cell cycle changes. Annexin V-FITC/PI assay was used to detect apoptotic changes; Western blot was used to detect the level of cyclin D1, CDK4, c-caspase-3, c-caspase-9 and cytochrome C protein levels in cytoplasm and mitochondria; JC-1 method was used to detect the mitochondrial membrane potential. Results The expression of Notch1 in PSFs transfected with Notch1 siRNA lentivirus was significantly lower than that in the negative control lentiviruses and the non-transfected PSFs(P＜0.05).The proliferation of PSFs decreased after Notch1 was down-regulated (P＜0.05);the percen-tage of G₀/G₁ phase increased(P＜0.05); the apoptotic rate increased(P＜0.05); the expression of c-caspase-3 and c-caspase-9 protein increased(P＜0.05); the mitochondrial membrane potential of the cells decreased significantly(P＜0.05); the level of cyclin D1 and CDK4 protein in cells decreased(P＜0.05); cytochrome C protein increased in cytoplasm and decreased in mitochondria(P＜0.05). Conclusions Down-regulation of Notch1 inhibits proliferation and induces apoptosis of PSF.

Key words: Notch1, fibroblast from pathological scars, mitochondrial membrane potential, caspase

CLC Number:

R619.6

SU Jiang-wei, YU Hui, DU Kun, WANG Jie, LIU Lin, CAI Jian. Down-regulation of Notch1 inhibits proliferation and induces apoptosis of fibroblasts from pathological scars[J]. Basic & Clinical Medicine, 2020, 40(3): 328-333.

References

[1]Liu DQ, Li XJ, Weng XJ. Effect of BTXA on inhibiting hypertrophic scar formation in a rabbit ear model[J]. Aesthetic Plas Surg, 2017, 41:1-8.
[2]Tang ZM, Zhai XX, Ding JC. Expression of mTOR/70S6K signaling pathway in pathological scar fibroblasts and the effects of resveratrol intervention[J]. Mol Med Rep, 2017, 15:2546-2550.
[3]Collesi C, Felician G, Secco I, et al. Reversible Notch1 acetylation tunes proliferative signaling in cardiomyocytes[J]. Cardiovasc Res, 2018, 114:103-122.
[4]Zhang HD, Jiang LH, Sun DW, et al. miR-30a inhibits the biological function of breast cancer cells by targeting Notch1[J]. Int J Mol Med, 2017, 40:1235-1242.
[5]Hrstka SC, Li X, Nelson TJ. NOTCH1-dependent nitric oxide signaling deficiency in hypoplastic left heart syndrome revealed through patient-specific phenotypes detected in bioengineered cardiogenesis[J]. Stem Cells, 2017, 35:1106-1119.
[6]Hsu YH, Lin YF, Chen CH, et al. Far infrared promotes wound healing through activation of Notch1 signaling[J]. J Mol Med, 2017, 95:1-11.
[7]Park JC, Chang IB, Ahn JH, et al. Nerve growth factor stimulates glioblastoma proliferation through notch1 receptor signaling[J]. J orean Neurosurg Soc, 2018, 61:441-449.
[8]Tang ZM, Zhai XX, Ding JC. Expression of mTOR/70S6K signaling pathway in pathological scar fibroblasts and the effects of resveratrol intervention[J]. Mol Med Rep, 2017, 15:2546-2550.
[9]Blain SW. Targeting p27 tyrosine phosphorylation as a modality to inhibit CDK4 and CDK2 and cause cell cycle arrest in breast cancer cells[J]. Oncoscience, 2018, 5:144-145.
[10]Pierozan P, Karlsson O. PFOS induces proliferation, cell-cycle progression, and malignant phenotype in human breast epithelial cells[J]. Arch Toxicol, 2018, 92:705-716.
[11]Gang D, Zheng X, Jiang P, et al. Notch1 suppresses prostate cancer cell invasion via the metastasis-associated 1-KiSS-1 metastasis-suppressor pathway[J]. Oncol Lett, 2017, 14:4477-4482.
[12]Lu CJ, He YF, Yuan WZ, et al. Dihydromyricetin-mediated inhibition of the Notch1 pathway induces apoptosis in QGY7701 and HepG2 hepatoma cells[J]. World J Gastroenterol, 2017, 23:6242-6251.
[13]Liu T, Wu X, Li Y, et al. RBFOX3 regulates the chemosensitivity of cancer cells to 5-fluorouracil via the pi3k/akt, emt and cytochrome-c/caspase pathways[J]. Cell Physiol Biochem, 2018, 46:1365-1380.
[14]Jiang S, Wang Q, Feng M, et al. C2-ceramide enhances sorafenib-induced caspase-dependent apoptosis via PI3K/AKT/mTOR and Erk signaling pathways in HCC cells[J]. Appl Microbiol Biotechnol, 2017, 101:1535-1546.
[15]Cheng J, Wu Q, Lv R, et al. MicroRNA-449a inhibition protects h9c2 cells against hypoxia/reoxygenation-induced injury by targeting the notch-1 signaling pathway[J]. Cell Physiol Biochem, 2018, 46:2587-2600.