2-甲氧基雌二醇对小鼠恶性黑色素瘤B16细胞增殖、侵袭和迁移能力的影响
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摘要
目的探讨2-甲氧基雌二醇(2-Methoxyestradiol,2-ME)对恶性黑色素瘤(malignant melanoma,MM)B16细胞增殖、侵袭及迁移的影响。方法将体外培养的小鼠MM B16细胞用不同浓度(10、20、40μmol/L)的2-ME处理。未经任何处理的小鼠MM B16细胞,记作B16-negative。倒置显微镜下观察细胞形态,平板克隆形成实验检测2-ME处理小鼠MM B16细胞2周后的克隆形成能力。Transwell方法观察药物在体外对细胞趋化侵袭能力的影响。观察药物在体外对细胞迁移能力的影响。细胞划痕实验观察经10μmol/L 2-ME处理MM B16细胞24h后划痕愈合程度。结果与对照组比较,不同浓度的2-ME处理MM B16细胞后克隆形成率差异有统计学意义(P<0.05)。与对照组比较,2-ME组细胞侵袭能力及迁移能力差异均有统计学意义(P<0.05)。划痕实验证实对照组的MM B16细胞经过24h后几乎迁移至所有的划痕区域,而实验组MM B16-10μmol/L的划痕区域仅有部分由MM B16细胞占有。结论 2-ME对小鼠MM B16细胞的增殖具有抑制作用,能够抑制MM B16细胞的克隆形成能力。2-ME可以抑制小鼠MM B16细胞的侵袭和迁移运动能力。
Objective To observe the effects of 2-methoxyestradiol(2-ME)on proliferation,invasion,migration of malignant melanoma(MM)B16cells.Methods MM B16 cells cultured in vitro were regarded as the control group.MM B16 cells were treated with different concentrations of 2-ME(10,20,40μmol/L).MM B16 cells were recorded in MM B16-negative cells without any treatment.Morphological changes of MM B16 cells were observed by inverted microscope after 48 h.The plate colony assay was used to testify the colony-formation ability of MM B16 cells treated with 2-methoxyestradiol for 2 weeks.The transwell assay was used to detect the capacity of invasion and migration of MM B16 cells treated with 2-methoxyestradiol in different concentrations.The wound healing assay was used to testify the capacity of migration of MM B16 cells treated with 2-ME at the concentrations of 10μmol/L for 24 h.Results Compared with the control group the colony-formation ability of MM B16 cells treated with 2-ME was significantly inhibited(P<0.05).The capacity of invasion and migration of MM B16 cells treated with 2-ME was inhibited compared with the control group(P<0.05).The cells in the control group almost migrated to all the scratches.Only some scratches areas were occupied by cells in the 10μmol/L experimental group.Conclusion 2-ME could inhibit the proliferation of MM B16 cell in a timedose-dependent manner.2-ME could inhibit the cloning ability of MM B16 cell.2-ME could inhibit the capacity of invasion and migration of MM B16 cell.
Objective To observe the effects of 2-methoxyestradiol(2-ME)on proliferation,invasion,migration of malignant melanoma(MM)B16cells.Methods MM B16 cells cultured in vitro were regarded as the control group.MM B16 cells were treated with different concentrations of 2-ME(10,20,40μmol/L).MM B16 cells were recorded in MM B16-negative cells without any treatment.Morphological changes of MM B16 cells were observed by inverted microscope after 48 h.The plate colony assay was used to testify the colony-formation ability of MM B16 cells treated with 2-methoxyestradiol for 2 weeks.The transwell assay was used to detect the capacity of invasion and migration of MM B16 cells treated with 2-methoxyestradiol in different concentrations.The wound healing assay was used to testify the capacity of migration of MM B16 cells treated with 2-ME at the concentrations of 10μmol/L for 24 h.Results Compared with the control group the colony-formation ability of MM B16 cells treated with 2-ME was significantly inhibited(P<0.05).The capacity of invasion and migration of MM B16 cells treated with 2-ME was inhibited compared with the control group(P<0.05).The cells in the control group almost migrated to all the scratches.Only some scratches areas were occupied by cells in the 10μmol/L experimental group.Conclusion 2-ME could inhibit the proliferation of MM B16 cell in a timedose-dependent manner.2-ME could inhibit the cloning ability of MM B16 cell.2-ME could inhibit the capacity of invasion and migration of MM B16 cell.
引文
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[13]胡彩霞,赵连梅,张国强,等.2-甲氧基雌二醇对恶性黑素瘤B16细胞株增殖与凋亡的影响[J].中华皮肤科杂志,2015,48(3):166-170.
[14]庞新,王鹏.低分子肝素对人乳腺癌细胞MCF-7侵袭抑制实验研究[J].河北医科大学学报,2013,34(6):657-660.
[2]Li X,Sun J,Hu S,et al.Icariin induced B16melanoma tumor cells apoptosis,suppressed tumor growth and metastasis[J].Iran J Public Health,2014,43(6):847-848.
[3]Ding L,Wang XQ,Zhang J,et al.Underlying mechanism of2-methoxyestradiol-induced apoptosis and growth arrest in SKOV3 human ovarian cancer cells[J].Eur Rev Med Pharmacol Sci,2015,19(11):2084-2090.
[4]高珊,徐勤.垂体转化肿瘤基因的生物学特性及其与恶性肿瘤发生发展的关系[J].临床荟萃,2012,27(17):1558-1561.
[5]DeSantis CE,Lin CC,Mariotto AB,et al.Cancer treatment and survivorship statistics,2014[J].CA Cancer J Clin,2014,64(4):252-271.
[6]Kim T,Amaria RN,Spencer C,et al.Combining targeted therapy and immune checkpoint inhibitors in the treatment of metastatic melanoma[J].Cancer Biol Med,2014,11(4):237-246.
[7]Hardy KM,Strizzi L,Margaryan NV,et al.Targeting nodal in conjunction with dacarbazine induces synergistic anticancer effects in metastatic melanoma[J].Mol Cancer Res,2015,13(4):670-680.
[8]Mourah S,LebbéC.Molecular alterations in melanoma and targeted therapies[J].Bull Cancer,2014,101(Suppl 2):S5-11.
[9]Moschos SJ,Pinnamaneni R.Targeted Therapies in Melanoma[J].Surg Oncol Clin N Am,2015,24(2):347-358.
[10]Song IS,Jeong YJ,Jeong SH,et al.Combination treatment with 2-methoxyestradiol overcomes bortezomib resistance of multiple myeloma cells[J].Exp Mol Med,2013,45(10):e50.
[11]Ma L,Li G,Zhu H,et al.2-Methoxyestradiol synergizes with sorafenib to suppress hepatocellular carcinoma by simultaneously dysregulating hypoxia-inducible factor-1and-2[J].Cancer Lett,2014,355(1):96-105.
[12]Duncan GS,Brenner D,Tusche MW,et al.2-Methoxyestradiol inhibits experimental autoimmune encephalomyelitis through suppression of immune cell activation[J].Proc Natl Acad Sci U S A,2012,109(51):21034-21039.
[13]胡彩霞,赵连梅,张国强,等.2-甲氧基雌二醇对恶性黑素瘤B16细胞株增殖与凋亡的影响[J].中华皮肤科杂志,2015,48(3):166-170.
[14]庞新,王鹏.低分子肝素对人乳腺癌细胞MCF-7侵袭抑制实验研究[J].河北医科大学学报,2013,34(6):657-660.