斑蝥酸钠对肝癌细胞侵袭、迁移影响的实验研究

Effects of Sodium Cantharidate on Invasion and Migration of Hepatocellular Carcinoma Cells

目的:探讨斑蝥酸钠对人肝癌SMMC-7721和HepG2细胞侵袭、迁移的影响。方法:斑蝥酸钠4、8、16、32、64μmol/L处理SMMC-7721和HepG2细胞24、48、72 h,采用CCK-8法检测SMMC-7721和HepG2细胞存活率。通过CCK-8试验确定斑蝥酸钠给药浓度为4、8、16μmol/L,并设置空白对照组,集落克隆法检测斑蝥酸钠对SMMC-7721和HepG2细胞的增殖抑制作用;倒置显微镜观察SMMC-7721和HepG2细胞形态学变化;划痕实验和Transwell小室法检测SMMC-7721和HepG2细胞的侵袭和迁移能力;Western Blot检测SMMC-7721和HepG2细胞中上皮标志蛋白E-钙黏蛋白(E-cadherin)、间质标志蛋白波形蛋白(Vimentin)、神经性钙黏蛋白(N-cadherin)的表达。结果:相较于空白对照组,斑蝥酸钠4、8、16μmol/L组细胞增殖、侵袭、迁移的抑制率明显升高,集落克隆显著减少,划痕损伤愈合的趋势明显减少,细胞皱缩发亮,数目减少失去了正常形态,E-cadherin蛋白表达上调,N-cadherin和Vimentin蛋白表达明显下调(P<0.05)。结论:斑蝥酸钠具有抑制肝癌细胞SMMC-7721和HepG2侵袭和迁移的作用。

Objective:To investigate the effects of sodium cantharidate on the invasion and migration of human hepatocellular carcinoma cell lines SMMC-7721 and HepG2.Methods:Sodium cantharidate(4,8,16,32,64μmol/L)were treated on SMMC-7721 and HepG2 cells for 24,48 and 72 h,and CCK-8 assay was used to determine the viability of SMMC-7721 and HepG2 cells.The concentrations of sodium cantharidate 4,8,16μmol/L were determined by CCK-8 assay,and blank control group was set.The proliferation inhibition of SMMC-7721 and HepG2 cells treated with sodium cantharidate was determined by colony cloning assay.The morphological changes of SMMC-7721 and HepG2 cells were observed by inverted microscope.The invasion and migration abilities of SMMC-7721 and HepG2 cells were detected by scratch assay and Transwell chamber assay.The expressions of epithelial marker protein E-cadherin,mesenchymal marker protein vimentin and neural cadherin in SMMC-7721 and HepG2 cells were determined by Western blot.Results:Compared with the blank control,sodium cantharidate groups(4,8,16μmol/L)significantly increased the inhibition rate of cell proliferation,invasion and migration,it also significantly decreased the colony clones and the tendency of scratch injury to heal,while the cells were observed shrunk and brighter,the number of cells was reduced,and morphological changes were observed.The protein expression level of E-cadherin was increased,while the protein levels of N-cadherin and vimentin were decreased significantly.Conclusion:Sodium cantharidate inhibited the invasion and migration of hepatoma cells SMMC-7721 and HepG2.