黄连素对体外培养肝癌细胞生物学行为的影响
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摘要
目的研究黄连素对体外培养肝癌HepG2细胞增殖、凋亡、迁移和侵袭的影响并探讨分子机制。方法不同浓度黄连素体外作用肝癌HepG2细胞后,MTT法检测增殖,流式细胞法检测凋亡,划痕法及Transwell法检测细胞迁移及侵袭,免疫印迹法检测MMP-9及VEGF蛋白表达。结果 12.5～200μmol/L黄连素均可对肝癌HepG2细胞产生明显的抑制增殖和诱导凋亡作用(P<0.05,P<0.01),浓度越高作用时间越长效果越好;50～200μmol/L黄连素还可抑制HepG2细胞迁移及侵袭并下调MMP-9和VEGF蛋白表达。结论黄连素可通过下调MMP-9以及VEGF蛋白表达对体外培养的肝癌HepG2细胞增殖、凋亡、迁移和侵袭等生物学行为产生影响。
Objective To study the effects of Berberine on biological behaviour of Liver Carcinoma Cell Line HepG2 cultured in vitro and explore its mechanism.Methods The growth and proliferation of HepG2 cell was detected by MTT assay. Cell apoptosis was detected by the flow cytometry. Cell invasive and migration ability was measured with Transwell assay and scratch adhesion test, respectively. The expression of MMP-9 and VEGF Protein was detected by Western blot.Results 12.5～200μmol/L of Berberine could inhibit cell proliferation and induce cell apoptosis in a time and concentration dependent manner(P<0.05,P<0.01), 50～200μmol/L of Berberine could inhibit HepG2 cell invasive and migration and markedly suppress MMP-9 and VEGF Protein.level. Conclusion Berberine could influence the biological behaviour of Liver Carcinoma Cell Line HepG2 cultured in vitro partly by down-regulating the level of MMP-9 and VEGF protein.
Objective To study the effects of Berberine on biological behaviour of Liver Carcinoma Cell Line HepG2 cultured in vitro and explore its mechanism.Methods The growth and proliferation of HepG2 cell was detected by MTT assay. Cell apoptosis was detected by the flow cytometry. Cell invasive and migration ability was measured with Transwell assay and scratch adhesion test, respectively. The expression of MMP-9 and VEGF Protein was detected by Western blot.Results 12.5～200μmol/L of Berberine could inhibit cell proliferation and induce cell apoptosis in a time and concentration dependent manner(P<0.05,P<0.01), 50～200μmol/L of Berberine could inhibit HepG2 cell invasive and migration and markedly suppress MMP-9 and VEGF Protein.level. Conclusion Berberine could influence the biological behaviour of Liver Carcinoma Cell Line HepG2 cultured in vitro partly by down-regulating the level of MMP-9 and VEGF protein.
引文
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[12] 孟燕,魏娟,曹风军.ERK通路激动剂EGF及抑制剂U0126对HepG2肝癌细胞迁移和侵袭能力的影响[J].解剖学研究,2016,38(6):452.
[13] 向龙超,李洪亮,曹风军,等.HMGB - 1 /RAGE 在肝癌增殖及转移中的作用研究进展[J].湖北医药学院学报,2016,35(1):101.
[14] 王娟,谢波,刘英,等.黄连素通过抑制PLCγ1 活性阻止肝癌细胞的侵袭[J].湖北医药学院学报,2015,34(1):23.
[15] Dung T D,Feng C C,Kuo W W,et al.Suppression of plasminogen activators and the MMP-2/-9 pathway by a Zanthoxylum avicennae extract to inhibit the HA22T human hepatocellular carcinoma cell migration and invasion effects in vitro and in vivo via phosphatase 2A activation[J].Biosci Biotechnol Biochem,2013,77(5):1814.
[2] WANG Ning,TAN Hor-yue.Berberine and Coptidis Rhizoma as potential anticancer agents:Recent updates and future perspectives[J].Journal of Ethnopharmacology,2015,176:35.
[3] 刘刚.黄连素抗癌特性的研究进展[J].广东医学院学报,2012,30( 2) :210.
[4] Lee SJ,Noh HJ,Sung EG,et al.Berberine sensitizes TRAIL-induced apoptosis through proteasome-mediated down regulation of c-FLIP and Mcl-1proteins[J].Int J Oncol,2011,38( 2) :485.
[5] 张春洁,金平,李娜.黄连素抑制人卵巢癌SKOV3 细胞增殖并诱导凋亡[J].基础医学与临床,2013,33 ( 2) :225.
[6] 雷博婷,马超英.黄连素抗肝癌的研究进展[J].中华中医药学刊,2014,32(2):254.
[7] 郑慧颖,吴献群,林丽琴,等.黄连素治疗妇科疾病的研究进展[J].时珍国医国药,2015,26(11):2751.
[8] 林庆新.黄连素诱导人肝癌HepG2细胞凋亡及其机制研究[J].中药材,2015,38(7):1499.
[9] 陈建卫,高娟,朱忠辉,等.黄连素对肝癌细胞系HepG2细胞体外迁移影响及其机制研究[J].陕西医学杂志,2018,47(6):686.
[10] 马玉娇,杨天枫,张彦民,等.黄连素联合塔斯品碱衍生物对肝癌细胞迁移及凋亡的影响[J].西北大学学报,2016,46(5):702.
[11] 尹作静,曹志伟,闫鑫淼,等.黄连素和吴茱萸碱协同抗癌的miRNA 网络机制研究[J].中国药理学通报,2017,33( 6) :772.
[12] 孟燕,魏娟,曹风军.ERK通路激动剂EGF及抑制剂U0126对HepG2肝癌细胞迁移和侵袭能力的影响[J].解剖学研究,2016,38(6):452.
[13] 向龙超,李洪亮,曹风军,等.HMGB - 1 /RAGE 在肝癌增殖及转移中的作用研究进展[J].湖北医药学院学报,2016,35(1):101.
[14] 王娟,谢波,刘英,等.黄连素通过抑制PLCγ1 活性阻止肝癌细胞的侵袭[J].湖北医药学院学报,2015,34(1):23.
[15] Dung T D,Feng C C,Kuo W W,et al.Suppression of plasminogen activators and the MMP-2/-9 pathway by a Zanthoxylum avicennae extract to inhibit the HA22T human hepatocellular carcinoma cell migration and invasion effects in vitro and in vivo via phosphatase 2A activation[J].Biosci Biotechnol Biochem,2013,77(5):1814.