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目的研究桑叶液对与乳腺癌肿瘤细胞(MDA-MB-231)共培养的人脐静脉内皮细胞(HUVEC-2C)增殖、迁移、凋亡及其分子表达的影响。方法用Transwell建立与肿瘤细胞共培养HUVEC-2C模型,实验分为单独培养HUVEC-2C组,细胞共培养组和添加高、中、低剂量桑叶液实验组,利用CCK-8细胞活性检测试剂盒检测与肿瘤共培养的HUVEC-2C的增殖情况;用24孔transwell迁移小板培养观察HUVEC-2C与肿瘤共培养的情况下,HUVEC-2C的迁移情况,实验组分为共培养组和高剂量组;通过流式细胞仪检测与肿瘤细胞共培养的HUVEC-2C凋亡状况;酶联免疫吸附测定(ELISA)法检测共培养体系中血管内皮生长因子(VEGF)分泌影响,实验分为共培养组和高、中、低剂量实验组。结果桑叶液对HUVEC-2C的增殖具有抑制作用,并随着桑叶液浓度的增加而抑制能力增强,在桑叶液浓度为0.1、0.05、0.025g/m L时,与共培养组相比较均具有统计学意义(P<0.05),当桑叶液浓度为0.1g/m L时,其对HUVEC-2C增殖抑制率较为明显;通过观察Transwell小室,并与共培养组相比较,添加了桑叶液的HUVEC-2C的迁移受到了一定的制约;流式细胞仪(FCM)检测到桑叶液能够促进HUVEC-2C凋亡;且能够抑制共培养体系中的VEGF的表达。结论桑叶能够抑制与肿瘤细胞共培养的HUVEC-2C的增殖、迁移以及促进HUVEC-2C的凋亡,达到抑制肿瘤血管生成的目的,其机制可能与抑制VEGF的表达有关。
Objective To study the effects of mulberry leaf decoction on the proliferation, migration, apoptosis and molecular expression of human umbilical vein endothelial cells (HUVEC-2C) co-cultured with breast cancer cells (MDA-MB-231). Methods The HUVEC-2C model was co-cultured with tumor cells using Transwell. The experiment was divided into HUVEC-2C group, cell co-culture group and high, medium and low dose mulberry leaf liquid group. CCK-8 cell activity assay reagent The proliferation of HUVEC-2C co-cultured with the tumor was detected by flow cytometry. The migration of HUVEC-2C was observed in 24-well transwell platelet transfection with HUVEC-2C co-cultured with tumor. The experimental groups were co-culture group and high Dose group. The apoptosis of HUVEC-2C cells co-cultured with tumor cells was detected by flow cytometry. The secretion of vascular endothelial growth factor (VEGF) in co-culture system was detected by enzyme linked immunosorbent assay (ELISA) Training group and high, medium and low dose experimental group. Results Mulberry leaf liquid had inhibitory effect on the proliferation of HUVEC-2C, and its inhibitory ability increased with the increase of Mulberry leaf liquid concentration. Compared with the co-culture group, the Mulberry leaf liquid concentration was 0.1, 0.05 and 0.025g / m L (P <0.05). When the mulberry leaf liquid concentration was 0.1g / m L, the inhibition rate of HUVEC-2C proliferation was more obvious. By observing the Transwell chamber and comparing with the co-culture group, The migration of HUVEC-2C in leaf liquid was restricted. Flow cytometry (FCM) detected that mulberry leaf fluid could promote the apoptosis of HUVEC-2C and inhibited the expression of VEGF in co-culture system. Conclusion Mulberry leaves can inhibit the proliferation and migration of HUVEC-2C co-cultured with tumor cells and promote the apoptosis of HUVEC-2C to inhibit tumor angiogenesis. The mechanism may be related to the inhibition of VEGF expression.