地鳖虫醇提物对肿瘤细胞侵袭与凋亡的作用机制

Mechanism of ethanol extract of ground turtle worm on tumor cell invasion and apoptosis

目的探究地鳖虫醇提物(ESWE)对前列腺癌细胞的迁移、侵袭、凋亡的影响及其作用机制。方法体外培养前列腺癌细胞,通过划痕实验测定其对细胞运动能力的影响,transwell小室做细胞迁移侵袭能力测定,吖啶橙溴乙锭(AO/EB)染色观察其对前列腺癌细胞凋亡的影响,Western印迹(WB)检测整合素(ITG)β1/ITG连接激酶(ILK)/纤维型肌动蛋白(F-actin)通路相关基因的表达。结果通过毒性试验得出ESWE对前列腺癌细胞的IC50值为6.265 mg/ml;随ESWE浓度增加,细胞运动能力、迁移与侵袭能力逐渐减弱;ITGβ1、ILK mRNA表达量呈递减趋势(P<0.05);半胱氨酸天冬氨酸蛋白酶(caspase)3增加表达,促进凋亡。骨架蛋白F-actin重构受阻并未影响F-actin mRNA的表达。结论ESWE通过抑制ITGβ1的表达从而调节F-actin重构,抑制PC3细胞迁移与侵袭,并上调其下游caspase3蛋白表达促进细胞凋亡。

Objective To study the effect of ethanol extract of earthworm(ESWE)on the migration,invasion and apoptosis of prostate cancer cells and its mechanism.Methods Prostate cancer cells were cultured in vitro,and their effects on cell motility were measured by scratch test.Transwell chambers were used for cell migration and invasion ability.AO/EB staining was used to observe their effects on prostate cancer cell apoptosis.WB was used to detect ITGβ1/ILK/F-actin pathway related gene expression.Results According to the toxicity test,the IC50 value of ESWE on prostate cancer cells was 6.265 mg/ml;with the increase of ESWE concentration,cell motility,migration and invasion ability gradually weakened;Integrinβ1,ILK mRNA expression showed a decreasing trend;the expression of caspase3 was increased and apoptosis was promoted,there were all significant differences(P<0.05).The remodeling of skeleton protein F-actin did not affect the expression of actin mRNA.Conclusions ESWE regulates the remodeling of F-actin by inhibiting the expression of ITGβ1,inhibits the migration and invasion of PC3 cells,and up-regulates the expression of the downstream caspase3 protein to promote cell apoptosis.