基于ciRS-7/miR-7/FAK通路研究掌叶半夏水提物对宫颈癌的抑制作用  

Inhibitory Effect of Pinellia Extract on Cervical Cancer Based on ciRS-7/miR-7/FAK Pathway

作　　者：王明喜[1] 张丽霞[1] 王长平 朱前勇 WANG Mingxi;ZHANG Lixia;WANG Changping;ZHU Qianyong(Anyang Hospital of traditional Chinese Medicine,Anyang Henan China 455000;Henan Provincial People′s Hospital,Zhengzhou Henan China 450000)

机构地区：[1]安阳市中医院,河南安阳455000 [2]河南省人民医院,河南郑州450000

出　　处：《中医学报》2022年第6期1278-1283,共6页Acta Chinese Medicine

基　　金：国家自然科学基金项目(8187081428)。

摘　　要：目的:探讨掌叶半夏水提物(pinellia extract,PE)通过调节小脑变性相关蛋白1反义转录物(circular RNA sponge for microRNA-7,ciRS-7)/microRNA-7(miR-7)/着斑激酶(focal adhesion kinase,FAK)通路对宫颈癌细胞的抑制作用。方法:体外培养人宫颈癌细胞(HeLa),分为HeLa细胞对照组和HeLa细胞PE干预组(0.15 g·L^(-1)),Transwell小室实验检测HeLa细胞侵袭及迁移能力。18只雌性BALB/c-nu裸小鼠随机分为正常对照组、皮下移植瘤模型组和PE干预组,每组6只。取对数生长期的人宫颈癌HeLa细胞,调整密度为1×10^(7) mL^(-1),除正常组外,其余组裸小鼠颈背部皮下注射接种人宫颈癌HeLa细胞0.2 mL建立移植瘤模型。当瘤体直径大约5 mm时,PE干预组裸鼠腹腔注射PE 10 mg·kg^(-1),其余组给予同等体积的生理盐水,每天1次,连续3周。检测小鼠的肿瘤体积,并计算抑瘤率;RT-qPCR法检测各组小鼠肿瘤组织中ciRS-7 mRNA及miR-7 mRNA的表达水平;Western Blot法检测各组小鼠肿瘤组织中FAK及p-FAK的蛋白表达水平。结果:与HeLa细胞对照组比较,HeLa细胞PE干预组HeLa细胞侵袭及迁移数目显著降低(P<0.01)。与正常对照组比较,皮下移植瘤模型组小鼠ciRS-7 mRNA、FAK蛋白表达水平显著升高(P<0.05),miR-7 mRNA、p-FAK蛋白表达水平显著降低(P<0.05);与皮下移植瘤模型组比较,PE干预组荷瘤裸鼠肿瘤体积明显变小(P<0.01),肿瘤质量明显降低(P<0.01),ciRS-7 mRNA、FAK蛋白表达水平显著降低(P<0.05),miR-7 mRNA、p-FAK蛋白表达水平显著升高(P<0.05)。结论:PE可抑制宫颈癌HeLa细胞的侵袭及迁移,并抑制宫颈癌肿瘤的生长,其机制可能与调节ciRS-7/miR-7/FAK信号通路有关。Objective:To investigate the inhibitory effect of Pinellia extract(PE)on cervical cancer cells by regulating the ciRS-7/miR-7/FAK pathway.Methods:Human cervical cancer cells(HeLa)were cultured in vitro and divided into HeLa cell control group and HeLa cell PE intervention group(0.15 mg·mL^(-1)).The invasion and migration ability of HeLa cells were detected by Transwell chamber invasion.18 female BALB/c-nu nude mice were randomly divided into normal control group,subcutaneous transplantation tumor model group,and PE intervention group(n=6).Human cervical cancer HeLa cells in the logarithmic growth phase were taken,and the concentration was adjusted to 1×10^(7) mL^(-1).Except for the normal group,nude mice in the other groups were inoculated with 0.2 mL of human cervical cancer HeLa cells subcutaneously on the back of the neck to establish a transplanted tumor model.When the tumor diameter was about 5 mm,the nude mice in the PE intervention group were intraperitoneally injected with PE 10 mg·kg^(-1),and the other groups were given the same volume of normal saline,once a day for 3 consecutive weeks.The tumor volume of the mice was detected,and the tumor inhibition rate was calculated.The expression level of ciRS-7/miR-7 in tumor tissues of mice in each group was detected by RT-qPCR.The expression levels of FAK and p-FAK protein in tumor tissues of each group were detected by Western blot.Results:Compared with the HeLa cell control group,the number of HeLa cell invasion and migration in the HeLa cell PE intervention group was significantly decreased(P<0.01).Compared with the normal control group,the expression levels of ciRS-7 mRNA and FAK protein in the subcutaneously transplanted tumor model group were significantly increased(P<0.05),while the expression levels of miR-7 mRNA and p-FAK protein were significantly decreased(P<0.05).Compared with the subcutaneously transplanted tumor model group,the tumor volume of the tumor-bearing nude mice in the PE intervention group was significantly smaller(P<0.01),the tumor m

关 键 词：宫颈癌 掌叶半夏水提物 ciRS-7/miR-7/FAK通路 小鼠 

分 类 号：R285.5[医药卫生—中药学]