Objective: To analyze and confirm whether scutebarbatine(SBT) has a certain inhibitory effect on osteosarcoma cells, and to preliminarily analyze its molecular mechanism in inhibiting osteosarcoma cells. Methods: Cell proliferation was detected by methyl thiazolyl tetrazolium(MTT) assay and colony formation assay. Apoptosis, migration and invasion ability of the cells were respectively determined by Hoechst 33258 staining, scratch test and by cell invasion and migration assays. Western blot was used to analyze the protein levels of apoptosis related molecules, including BAD, BCL2 and cleaved caspase 3. The luciferase activity of TopLuc was detected by luciferase reporter gene assay. Western blot was used to detect the expression of β-Catenin, proto-oncogene(C-myc), G1/s-specific cyclin D1, glycogen synthase kinase-3β(GSK3β) and the phosphorylation of serine 9(Ser9) of GSK3 β. Results: SBT could inhibit the proliferation, migration and invasion of osteosarcoma cells, promote apoptosis of osteosarcoma cells and inhibit BCL protein level, but it upregulated BAD and cleaved caspase 3 protein levels. SBT inhibited the activity of TopLuc luciferase and Ser9, down-regulated the protein levels of β-catenin and its target molecules, C-myc and cyclin D1, which ultimately led to Wnt/β-catenin signal inactivated. Conclusion: SBT can inhibit the proliferation, migration and invasion of human osteosarcoma cells, and promote the apoptosis of osteosarcoma cells, which may be related to the inhibition of Wnt/β-Catenin signaling pathway by SBT. |
[1] MEAZZA C, SCANAGATTA P.Metastatic osteosarcoma:a challenging multidisciplinary treatment[J]. Expert Rev Anticancer Ther, 2016, 16(5):543-556.
[2] SCHUETZE S M.Chemotherapy in the management of osteosarcoma and Ewing's sarcoma[J]. J Natl Compr Canc Netw, 2007, 5(4):449-455.
[3] FERRARI S, SERRA M.An update on chemotherapy for osteosarcoma[J]. Expert Opin Pharmacother, 2015, 16(18):2727-2736.
[4] YAMAMOTO N, TSUCHIYA H.Chemotherapy for osteosarcoma-where does it come from? What is it? Where is it going?[J]. Expert Opin Pharmacother, 2013, 14(16):2183-2193.
[5] RASTOGI S, AGGARWAL A, TIWARI A, et al. Chemotherapy in nonmetastatic osteosarcoma:recent advances and implications for developing countries[J]. J Glob Oncol, 2018, 4:1-5.
[6] BISHOP M W, JANEWAY K A, GORLICK R.Future directions in the treatment of osteosarcoma[J]. Curr Opin Pediatr, 2016, 28(1):26-33.
[7] LEE S R, KIM M S, KIM S, et al. Constituents from scutellariabarbata inhibiting nitric oxide production in LPS-stimulated microglial cells[J]. Chem Biodivers, 2017, 14(11):e1700231.
[8] 史长灿, 赵瑞芝, 卢传坚.半枝莲中总生物碱的提取及抑菌作用的初步研究[J]. 中成药, 2013, 35(6):1315-1319.
[9] YANG X K, XU M Y, XU G S, et al. In vitro and in vivo antitumor activity of scutebarbatine on human lung carcinoma A549 cell lines[J]. Molecules, 2014, 19(7):8740-8751.
[10] LI Y Y, TANG X L, JIANG T, et al. Bioassay-guided isolation of neo-clerodanediterpenoids from Scutellariabarbata[J]. J Asian Nat Prod Res, 2013, 15(9):941-949.
[11] 杨沙, 段灿灿, 晏仁义, 等.基于网络药理学的半枝莲抗肿瘤活性成分及整合作用机制研究[J]. 中草药, 2018, 49(15):3471-3482.
[12] JULIEN O, WELLS J A.Caspases and their substrates[J]. Cell Death Differ, 2017, 24(8):1380-1389.
[13] YOULE R J, STRASSER A.The BCL-2 protein family:opposing activities that mediate cell death[J]. Nat Rev Mol Cell Biol, 2008, 9(1):47-59.
[14] MARTINS-NEVES S R, CORVER W E, PAIVA-OLIVEIRA D I, et al. Osteosarcoma stem cells have active Wnt/β-catenin and overexpress SOX2 and KLF4[J]. J Cell Physiol, 2016, 231(4):876-886.
[15] TIAN J, HE H, LEI G.Wnt/β-catenin pathway in bone cancers[J]. Tumour Biol, 2014, 35(10):9439-9445.
[16] XIE D, ZHENG G Z, XIE P, et al. Antitumor activity of resveratrol against human osteosarcoma cells:a key role of Cx43 and Wnt/β-catenin signaling pathway[J]. Oncotarget, 2017, 8(67):111419-111432.
[17] TAN T, CHEN J, HU Y, et al. Dihydrotanshinone I inhibits the growth of osteosarcoma through the Wnt/β-catenin signaling pathway[J]. Onco Targets Ther, 2019, 12:5111-5122.
[18] ZHANG Z F, WANG Y J, FAN S H, et al. MicroRNA-182 downregulatesWnt/β-catenin signaling, inhibits proliferation, and promotes apoptosis in human osteosarcoma cells by targeting HOXA9[J]. Oncotarget, 2017, 8(60):101345-101361.
[19] YUSKAITIS C J, BEUREL E.Glycogen synthase kinase-3(GSK3):inflammation, diseases, and therapeutics[J]. Neurochem Res, 2007, 32(4-5):577-595. |