目的 研究川芎嗪调节Yes相关蛋白(YAP)/转录辅激活因子(TAZ)信号通路对宫颈癌细胞上皮间质转化和放疗耐药性的影响.方法 将人宫颈癌细胞株C33A随机分为对照组(不做任何干预处理)、川芎嗪组(1.5 mg·mL-1川芎嗪)、川芎嗪+空载组(1.5 mg·mL-1 川芎嗪+转染空载质粒)、川芎嗪+YAP1过表达组(1.5 mg·mL-1川芎嗪+转染YAP1过表达质粒).构建人宫颈癌放射线抵抗细胞C33A-RR并随机分为放射抵抗对照组(不做任何干预处理)、放射组(6 Gy射线)、放射+川芎嗪组(6 Gy射线+1.5 mg·mL-1川芎嗪)、放射+川芎嗪+空载组(6 Gy射线+1.5 mg·mL-1川芎嗪+转染空载质粒)、放射+川芎嗪+YAP1过表达组(6 Gy射线+1.5 mg·mL-1川芎嗪+转染YAP1过表达质粒).用蛋白质印迹法检测蛋白相对表达水平;用细胞划痕和Transwell侵袭实验分别检测细胞迁移和侵袭情况;用细胞计数试剂盒8(CCK-8)检测细胞增殖情况.结果 对照组、川芎嗪组、川芎嗪+空载组和川芎嗪+YAP1过表达组的迁移率分别为(84.82±12.16)％、(20.67±4.48)％、(21.22±5.03)％和(76.74±0.15)％,E-cadherin 蛋白相对表达水平分别为 0.16±0.03、0.70±0.08、0.72±0.13和0.19±0.04,川穹嗪组上述指标与对照组比较,在统计学上差异均有统计学意义(均P＜0.05);川芎嗪+YAP1过表达组上述指标与川芎嗪组、川芎嗪+空载组比较,在统计学上差异均有统计学意义(均P＜0.05).放射抵抗对照组、放射组、放射+川芎嗪组、放射+川芎嗪+空载组和放射+川芎嗪+YAP1过表达组YAP1蛋白相对表达水平分别为0.79±0.14、0.88±0.16、0.21±0.03、0.22±0.04 和 0.82±0.16,细胞的存活率分别为(100.00±0.00)％、(95.78±20.12)％、(40.13±6.07)％、(42.21±6.45)％和(90.12±18.65)％.放射组上述指标与放射抵抗对照组比较,在统计学上差异均有统计学意义(均P＜0.05);放射+川芎嗪组、放射+川芎嗪+空载组上述指标与放射组、放射+川芎嗪+YAP1过表达组比较,在统计学上差异均有统计学意义(均P＜0.05).结论 川芎嗪可下调YAP/TAZ信号通路蛋白相对表达水平,从而抑制宫颈癌细胞上皮间质转化、迁移和侵袭,并降低其放疗耐药性,促进其凋亡.
Objective To study the impacts of tetramethylpyrazine(TMP)on the epithelial mesenchymal transformation and radiotherapy resistance of cervical cancer cells by regulating Yes-associated protein(YAP)/transcriptional coactivator(TAZ)signal pathway.Methods Human cervical cancer cell line C33A was randomly grouped into control group(without any intervention),TMP group(1.5 mg·mL-1TMP),TMP+NC group(TMP 1.5 mg·mL-1+transfection empty plasmid)and TMP+YAP1 group(TMP 1.5 mg·mL-1+transfection YAP1 overexpression plasmid).C33A-RR cells of human cervical cancer were constructed and randomly separated into control-RR group(without any intervention),radiation group(6 Gy radiation),RT group(6 Gy radiation+TMP 1.5 mg·mL-1),RTN group(6 Gy radiation+TMP 1.5 mg·mL-1+transfection empty plasmid)and RTY group(6 Gy radiation+TMP 1.5 mg·mL-1+transfection YAP1 overexpression plasmid).Western blot detected the relative expression levels of proteins;cell scratch and Transwell invasion experiments respectively examined cell migration and invasion;cell counting kit-8(CCK-8)and flow cytometry experiments respectively evaluated cell proliferation and apoptosis.Results The mobility of control group,TMP group,TMP+NC group and TMP+YAP1 group were(84.82±12.16)％,(20.67±4.48)％,(21.22±5.03)％and(76.74±0.15)％,respectively;E-cadherin protein expression levels were 0.16±0.03,0.70±0.08,0.72±0.13 and 0.19±0.04.Compared TMP group with control group of above indictors were statistically significant(all P＜0.05);compared TMP+YAP1 group with TMP group and TMP+NC group were statistically significant(all P＜0.05).YAP1 protein expression levels in control-RR group,radiation group,the RT group,RTN group and RTY group were 0.79±0.14,0.88±0.16,0.21±0.03,0.22±0.04 and 0.82±0.16,respectively;the survival rates were(100.00±0.00)％,(95.78±20.12)％,(40.13±6.07)％,(42.21±6.45)％and(90.12±18.65)％,respectively.The difference between radiation group and control-RR group were statistically significant(all P＜0.05);the differences between RT group,RTN group and radiation group,RTY group were statistically significant(all P＜0.05).Conclusion TMP can down regulate the expression of YAP/TAZ signal pathway protein,thereby inhibiting epithelial mesenchymal transformation,migration and invasion of cervical cancer cells,reducing their resistance to radiotherapy and promoting their apoptosis.
郭宗梁;张彩霞;张新悦
内蒙古医科大学附属医院妇产科,内蒙古自治区呼和浩特 010059
中医学
川芎嗪;Yes相关蛋白/转录辅激活因子;宫颈癌;上皮间质转化;放疗耐药性
tetramethylpyrazine;Yes-associated protein/transcriptional coactivator;cervical cancer;epithelial mesenchymal transformation;radiotherapy resistance
《中国临床药理学杂志》 2024 (008)
1165-1169 / 5
10.13699/j.cnki.1001-6821.2024.08.016
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