根皮素调节YAP/TAZ信号通路对HPV阳性宫颈癌细胞恶性进展的影响

Effect of phloretin on the malignant progression of HPV positive cervical cancer cells by regulating the YAP/TAZ signaling pathway

目的探讨根皮素调节含有PDZ结合位点转录共激活因(YAP/TAZ)信号通路对HPV阳性宫颈癌细胞恶性进展的影响。方法体外培养HPV16阳性宫颈癌SiHa细胞并分为根皮素低(40μg/mL)、中(80μg/mL)、高剂量组(160μg/mL)、激活剂组(160μg/mL根皮素+YAP激活剂(PY-60)10μmol/L)、抑制剂组(160μg/mL根皮素+YAP抑制剂(VTPF)5μmol/L)和对照组。CCK-8实验检测细胞增殖;流式细胞术检测细胞凋亡;划痕试验检测细胞迁移;Transwell检测细胞侵袭;Western blot检测细胞YAP/TAZ信号通路及凋亡蛋白表达。结果相较于对照组,根皮素低、中、高剂量组A450值、细胞侵袭数目、划痕愈合率及YAP、TAZ、B淋巴细胞瘤-2(Bcl-2)表达下降,凋亡率和Bcl-2相关X蛋白(Bax)表达增高,且各剂量组间差异显著(P<0.05)。相较于根皮素高剂量组,激活剂组A450值、细胞侵袭数目、划痕愈合率及YAP、TAZ、Bcl-2表达增加,凋亡率和Bax表达下降(P<0.05),抑制剂组各指标水平相反。结论根皮素可抑制HPV阳性宫颈癌细胞迁移、侵袭和增殖,其机制可能与下调YAP/TAZ信号通路有关。

Objective To investigate the impact of phloretin on the malignant progression of HPV positive cervical cancer cells by regulating the Yes-associated protein/transcriptional coactivator with PDZ-binding motif(YAP/TAZ)signaling pathway.Methods HPV16 positive cervical cancer SiHa cells were cultured in vitro and separated intoa low-dose phloretin group(40μg/mL),a medium-dose phloretin group(80μg/mL),a high-dose phloretin group(160μg/mL),an activator group[160μg/mL phloretin+YAP activator(PY-60)10μmol/L],and an inhibitor group[160μg/mL Phloretin+YAP inhibitor(VTPF)5μmol/L]and a control group.CCK-8 experiment was applied to detect cell proliferation.Flow cytometry was applied to detect cell apoptosis.Scratch test was applied to detect cell migration.Invasion was detected by Transwell.The expression of YAP/TAZ signaling pathway and apoptotic proteins in cells were detected by Western blot.Results Compared with the control group,the A_(450)value,number of cell invasions,scratch healing rate,the expression of YAP,TAZ,and B cell lymphocyte tumor 2(Bcl-2)were increased in the low-dose,medium-dose,and high-dose phloretin groups,the cell apoptosis rate and the expression of Bcl-2 associated X protein(Bax)were increased,and there were significant differences among each dose group(P<0.05).Compared with the high-dose phloretin group,the A_(450)value,number of cell invasions,scratch healing rate,the expression of YAP,TAZ,and Bcl-2 in the activator group were obviously increased,the cell apoptosis rate and the expression of Bax were reduced(P<0.05),the level of the above indicators in the inhibitor group was opposite.Conclusion Phloretin can inhibit the migration,invasion andproliferation of HPV positive cervical cancer cells,and its mechanism may be related to the down-regulation of YAP/TAZ signaling pathway.