三氧化二砷对肝癌HepG2细胞形成血管生成拟态的影响及其机制

Influence of arsenic trioxide in vasculogenic mimicry of HepG2 cells and its mechanism

目的:探讨三氧化二砷(AS2O3)对肝癌HepG2细胞血管生成拟态(VM)形成的影响,初步阐明AS2O3对VM抑制的可能作用机制。方法:应用CCK-8法测定AS2O3对HepG2细胞作用72h的半数抑制浓度(IC50);以Matrigel胶体外三维培养HepG2细胞,将其分为空白对照组、1/2IC50AS2O3组和IC50AS2O3组,IPP软件计算VM的数量、长度和面积,Western blotting法检测VM相关蛋白VE-cadherin、基质金属蛋白酶2(MMP-2)、凋亡相关蛋白含半胱氨酸的天冬氨酸蛋白水解酶3(caspase-3)和增殖细胞核抗原(PCNA)的表达。结果:AS2O3作用HepG2细胞72h后IC50为10μmol·L-1。与对照组比较,1/2IC50AS2O3组和IC50 AS2O3组VM数量、长度和面积明显减少(P<0.01);与1/2IC50AS2O3组比较,IC50AS2O3组VM数量、长度和面积亦明显减少(P<0.05);与对照组比较,1/2IC50AS2O3组和IC50AS2O3组VE-cadherin和MMP-2蛋白表达水平降低(P<0.05),而caspase-3和PCNA蛋白表达水平无明显变化(P>0.05)。结论:AS2O3抑制HepG2细胞形成VM,其作用机制可能与抑制拟态通路相关蛋白VE-cadherin和MMP-2的表达有关。

Objective To explore the expression of caspase-3in skeletal muscle of the mice in the state of cancer,and to elucidate the relationship between Caspase-3and apoptosis,consumption of skeletal muscle protein in cancer cachexia.Methods 48male BALB/c mice were randomly divided into cancer cachexia group and control group（n=24）.The mice in cancer cachexia group were inoculated with mouse colon 26adenocarcinoma cells.The body weights of the mice in two groups were detected daily.Eight mice in each group were executed to test the weight of left gastrocnemius,fiber crosscut area,the expression levels of tumor necrosis factor-α（TNF-α）,interleukin-6（IL-6）,Caspase-3proteins and the apoptotic rate of gastrocnemius cells on day 8,14,and 20,respectively.Results The mice in cancer cachexia group appeared cachectic symptoms on day 14,the non-tumor body weight was decreased more than 20%of that in control group（P〈0.05）.Compared with control group at the same time,the mouse body weight,non-tumor body weight,the weight of left gastrocnemius and the fiber crosscut area of the mice in cancer cachexia group were obviously decreased with the prolongation of inoculation time（P〈0.05）,whereas the expression levels of TNF-α,IL-6,Caspase-3proteins and the apoptotic rate of muscle cells were obviously increased after tumor inoculation（P〈0.05）.The level of Caspase-3protein was negatively correlated with the weight of gastrocnemius and fiber crosscut area（r=-0.716,P〈0.05;r=-0.694,P〈0.05）,and the level of Caspase-3was positively correlated with the levels of TNF-αand IL-6（r=0.742,P〈0.05;r=0.675,P〉0.05）.Conclusion Caspase-3may be a key factor in the protein comsumption of skeletal muscle in cancer cachexia.