PG对宫颈癌HeLa细胞侵袭、转移和凋亡的影响

EFFECTS OF PG ON INVASION,METASTASIS AND APOPTOSIS OF CERVICAL CANCER HELA CELLS

目的探讨大黄素甲醚-8-O-β-葡萄糖苷(PG)对宫颈癌HeLa细胞侵袭、转移及凋亡的影响。方法采用一系列PG浓度(0、2.5、5、10、20、40、80、160、320μg/ml)作用HeLa细胞24 h并采用MTT法检查HeLa细胞活性情况,采用Transwell法与划痕试验、免疫荧光法、流式细胞法检测PG(20、40、80μg/ml)作用下HeLa细胞侵袭、转移、凋亡情况,采用Western blot检测细胞侵袭、转移及凋亡相关蛋白水平;构建体内HeLa细胞移植瘤裸鼠模型,PG(40 mg/kg)干预30 d,观察存活率、瘤组织凋亡率及其相关蛋白水平。结果在一定PG浓度范围内,HeLa细胞活力随着PG浓度增加而逐渐降低;PG(20、40、80μg/ml)作用下HeLa细胞的侵袭率、迁移率、Vimentin阳性细胞数目均显著低于0μg/ml PG处理(P<0.05),而细胞凋亡率显著高于0μg/ml PG处理(P<0.05);Western blot实验结果显示PG(20、40、80μg/ml)作用下HeLa细胞的MMP-9、N-cadherin、Bcl-2蛋白表达量低于0μg/ml PG处理(P<0.05),E-cadherin、Caspase-3、Caspase-9、Bax蛋白水平高于0μg/ml PG处理(P<0.05);PG(40mg/kg)处理组裸鼠的凋亡率、生存率、E-cadherin、Caspase-3显著高于对照组(P<0.05),裸鼠肿瘤组织质量、N-cadherin蛋白水平显著低于对照组(P<0.05)。结论 PG对HeLa细胞侵袭、转移具有抑制作用,对HeLa细胞凋亡具有促进作用,可能与调节侵袭转移、凋亡相关蛋白表达有关。

Objective To explore the effects of physcion 8-O-β-glucopyranoside(PG) on the invasion,metastasis and apoptosis of cervical cancer HeLa cells.Methods HeLa cells were treated with different concentration of PG(0,2.5,5,10,20,40,80,160,320 μg/ml) for 24 h.The viability of HeLa cell was detected by MTT.The invasion,metastasis and apoptosis of HeLa cells treated with PG(20,40,80 μg/ml) were detected by Transwell,scratch test,immunofluorescence and flow cytometry.The levels of cell invasion,metastasis and apoptosis-related proteins were detected by Western blot.The nude mouse models with transplanted tumors were constructed,which were intervened with PG(40 mg/kg) for 30 d.The survival rate,apoptosis rate of tumor tissues and levels of related proteins were observed.Results Within certain range of PG concentration,viability of HeLa cells was gradually decreased with the increase of PG concentration.The invasion rate,migration rate and number of Vimentin-positive HeLa cells treated with PG(20,40,80 μg/ml) were significantly lower than those treated with 0 μg/ml PG(P<0.05),while apoptosis rate was significantly higher than that treatedwith 0 μg/ml PG(P<0.05).Western blot results showed that expression levels of MMP-9,N-cadherin and Bcl-2 proteins in HeLa cells treated with PG(20,40,80 μg/ml) were lower than those treated with 0 μg/ml PG(P<0.05),while levels of E-cadherin,Caspase-3,Caspase-9 and Bax proteins were higher than those treated with 0 μg/ml PG(P<0.05).The apoptosis rate,survival rate,E-cadherin and Caspase-3 in PG(40 mg/kg) treatment group were significantly higher than those in control group(P<0.05),while mass of tumor tissue and level of N-cadherin protein was significantly lower than that in control group(P<0.05).Conclusion PG can inhibit the invasion and metastasis of HeLa cells,and promote their apoptosis,which may be related to regulating the expression of invasion,metastasis and apoptosis-related proteins.