茶溴香酰胺脂质体对黑色素瘤细胞生长和迁移的抑制作用
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摘要
考察茶溴香酰胺脂质体(TBrC-L)对黑色素瘤B16细胞生长和迁移的抑制作用及其分子机制,为研发抗癌新药提供科学依据.采用MTT法和细胞划痕术探究TBrC-L对黑色素瘤B16细胞生长和迁移的影响; Hoechst 33258染色法观察药物作用下细胞凋亡形态的变化; Western Blotting检测TBrC-L对B16细胞生长、迁移和凋亡相关蛋白表达的影响.结果表明:TBrC-L能对B16细胞的生长和迁移产生显著性抑制作用,并且诱导其凋亡; TBrC-L可下调c-Met、HGF、Bcl-2、NF-κB和VEGFR2的表达,上调Caspase-3、E-cadherin、Bax、p53和Cytochrome c的表达,抑制HGF/Met/VEGFR2-NF-κB通路可能是TBrC-L抑制黑色素瘤B16细胞生长和迁移作用的重要分子机制.本研究结果提示,TBrC-L具有进一步开发为抗黑色素瘤药物的潜力.
The purpose of this study is to investigate the inhibitory effect of theanine derivative TBrC-prepared liposomes( TBrC-L) on the growth and migration of melanoma B16 cells. The effects of different concentrations of TBrC-L on the growth and migration of melanoma B16 cells are investigated by using MTT and cell scratch tests respectively. Hoechst 33258 staining is used to observe the change of cell morphology under the action of drugs.Western blotting is used to analyze the related protein expressions in the melanoma cells. TBrC-L significantly inhibits the growth and migration of the melanoma B16 cells and induces the apoptosis in the melanoma cells. TBrCL significantly reduces the protein expressions of c-Met,HGF,Bcl-2,NF-κB and VEGFR2,while it remarkably enhances the protein expressions of caspase-3,E-cadherin,Bax,PARP,p53 and cytochrome c in B16 cells. The molecular mechanism of TBrC-L action may be associated with the inhibition of HGF/Met/VEGFR2-NF-κB signaling pathway. The results of this study indicate that TBrC-L has the potential for further development of anti-melanoma drugs.
The purpose of this study is to investigate the inhibitory effect of theanine derivative TBrC-prepared liposomes( TBrC-L) on the growth and migration of melanoma B16 cells. The effects of different concentrations of TBrC-L on the growth and migration of melanoma B16 cells are investigated by using MTT and cell scratch tests respectively. Hoechst 33258 staining is used to observe the change of cell morphology under the action of drugs.Western blotting is used to analyze the related protein expressions in the melanoma cells. TBrC-L significantly inhibits the growth and migration of the melanoma B16 cells and induces the apoptosis in the melanoma cells. TBrCL significantly reduces the protein expressions of c-Met,HGF,Bcl-2,NF-κB and VEGFR2,while it remarkably enhances the protein expressions of caspase-3,E-cadherin,Bax,PARP,p53 and cytochrome c in B16 cells. The molecular mechanism of TBrC-L action may be associated with the inhibition of HGF/Met/VEGFR2-NF-κB signaling pathway. The results of this study indicate that TBrC-L has the potential for further development of anti-melanoma drugs.
引文
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[2] IVES N,STOWE R,LORIGAN P,et al. Chemotherapy compared with biochemotherapy for the treatment of metastatic melanoma:A meta-analysis of 18 trials involving 2,621 Patients[J]. Journal of Clinical Oncology,2007,25(34):5426-5434.
[3] ASCIERTO P A,AGARWALA S S,CILIBERTO G,et al. Future perspectives in melanoma research"Melanoma Bridge",Napoli,November 30th-3rd December 2016[J]. Journal of Translational Medicine,2017,15(1):226-266.
[4] FRIEDMAN M,MACKEY B E,KIM H C,et al. Structure-activity relationships of tea compounds against human cancer cells[J]. Journal of Agricultural and Food Chemistry,2007,55(2):243-253.
[5]闫春燕,刘辉,朱荣芹,等.茶氨酸衍生物茶溴香酰胺脂质体对人肺癌细胞生长和迁移的抑制作用[J].安徽农业大学学报,2018,45(1):15-24.
[6]郑晓慧,朱荣芹,刘真真,等.茶氟香酰胺对人肝癌细胞生长和迁移的抑制作用[J].烟台大学学报(自然科学与工程版),2018,31(2):136-144.
[7] YU P F,LIU Q,LIU K,et al. Matrine suppresses breast cancer cell proliferation and invasion via VEGF-Akt-NF-κB signaling[J]. Cytotechnology,2009,59(3):219-229.
[8] LUAN J L,DUAN H Y,LIU Q,et al. Inhibitory effects of norcantharidin against human lung cancer cell growth and migration[J]. Cytotechnology,2010,62(4):349-355.
[9] HUH C,FACTOR V M,SANCHEZ A,et al. Hepatocyte growth factor/c-met signaling pathway is required for efficient liver regeneration and repair[J]. Proceedings of the National Academy of Sciences of the United States of America,2004,101(13):4477-4482.
[10] ZHANG J Y,GREEN C L,TAO S,et al. NF-κB Rel A opposes epidermal proliferation driven by TNFR1 and JNK[J].Genes&Development,2004,18(1):17.
[11] JOVE R. STATs in cancer inflammation and immunity:a leading role for STAT3[J]. Nature Reviews Cancer,2009,9(11):798-809.
[12] PARK S,LEE M,SON B,et al. TBK1-targeted suppression of TRIF-dependent signaling pathway of Toll-like receptors by 6-shogaol,an active component of ginger[J]. Bioscience,Biotechnology,and Biochemistry,2009,73(7):1474-1478.
[13] DE GIORGI F,LARTIGUE L,BAUER M K,et al.The permeability transition pore signals apoptosis by directing Bax translocation and multimerization[J]. The FASEB Journal,2002,16(6):607-609.
[14] SHALTIEL I A,APRELIA M,SAURIN A T,et al.Distinct phosphatases antagonize the p53 response in different phases of the cell cycle[J]. Proceedings of the National Academy of Sciences of the United States of America,2014,111(20):7313-7318.
[15]李文娟,潘庆杰,李美玉. p53基因及其功能研究进展[J].生物技术通讯,2014,25(2):282-285.
[16] KANG Q,ZOU H,YANG X L,et al. Characterization and prognostic significance of mortalin,Bcl-2 and Bax in intrahepatic cholangiocarcinoma[J]. Oncology Letters,2017,15(2):2161-2168.
[17] LIANG S H,SUN K,WANG Y,et al. Role of Cyt-C/caspases-9,3,Bax/Bcl-2 and the FAS death receptor pathway in apoptosis induced by zinc oxide nanoparticles in human aortic endothelial cells and the protective effect by alpha-lipoic acid[J]. Chemico-Biological Interactions,2016,258:40-51.
[18] HUANG W F,YANG Y C,LIU H,et al. Changes of Caspase-3 and Caspase-9 activity during the simvastatin-induced apoptosis in K562 cells[J]. Cancer Res Prevent Treatment,2007,34(1):39-41.
[19] ZHANG Y,JAIN R K,ZHU M,et al. Recent progress and advances in HGF/MET-Targeted therapeutic agents for cancer treatment[J]. Biomedicines,2015,3(1):149-181.
[20] ZENG Q H,CHEN S Q,YOU Z B,et al. Hepatocyte growth factor inhibits anoikis in head and neck squamous cell carcinoma cells by activation of ERK and Akt signaling independent of NF-κB[J]. Journal of Biological Chemistry,2002,277(28):25203-25208.
[21] GHERARDI E,BIRCHMEIER W,BIRCHMEIER C,et al. Targeting MET in cancer:rationale and progress[J]. Nature Reviews Cancer,2012,12(2):89-103.
[22] LIU B D,SUN L J,LIU Q,et al. A Cytoplasmic NF-κB interacting long noncoding RNA blocks IκB phosphorylation and suppresses breast cancer metastasis[J]. Cancer Cell,2015,27(3):370-381.
[23] LU Y J,XU Q,ZUO Y Z,et al. Isoprenaline/β2-AR activates Plexin-A1/VEGFR2 signals via VEGF secretion in gastric cancer cells to promote tumor angiogenesis[J]. BMC Cancer,2017,17(1):875.
[24] WHEELER K C,JENA M K,PRADHAN B S,et al.VEGF may contribute to macrophage recruitment and M2 polarization in the decidua[J]. PLOS ONE,2018,13(1):1-18.
[25] BROUXHON S M,KYRKANIDESY S,TENG X,et al.Soluble E-cadherin:a critical oncogene modulating receptor tyrosine kinases,MAPK and PI3K/Akt/m TOR signaling[J].Oncogene,2014,33(2):225-235.
[26] PETROVA Y I,SCHECTERSON L,GUMBINER B M,et al. Roles for E-cadherin cell surface regulation in cancer[J].Molecular Biology of the Cell,2016,27(21):3233-3244.
[27] BROUXHON S M,KYRKANIDES S,TENG X,et al.Soluble E-cadherin:a critical oncogene modulating receptor tyrosine,MAPK and PI3K/AKT/m TOR signaling[J]. Oncogene,2014,33(2):225-235.