丙泊酚通过miR-802/PTCH1/SHH通路抑制宫颈癌细胞迁移和侵袭的研究  

Inhibition of Propofol on Migration and Invasion of Cervical Cancer Cells Through miR-802/PTCH1/SHH Pathway

作　　者：罗江辉[1] 毛远舟[1] 张可贤[1] 邹江[1] LUO Jianghui;MAO Yuanzhou;ZHANG Kexian;ZOU Jiang(Dept.of Anesthesiology,Sichuan Cancer Hospital,Chengdu 610041,China)

机构地区：[1]四川省肿瘤医院麻醉科,成都610041

出　　处：《中国医院用药评价与分析》2022年第10期1164-1171,共8页Evaluation and Analysis of Drug-use in Hospitals of China

基　　金：四川省卫生健康委员会科研课题(No.19PJ081)。

摘　　要：目的:探讨丙泊酚通过miR-802/PTCH1/SHH通路对宫颈癌细胞迁移和侵袭的影响。方法:使用不同浓度丙泊酚处理人宫颈癌细胞HeLa为丙泊酚组,使用二甲基亚砜处理细胞为对照组。采用噻唑蓝实验和Transwell实验分别检测HeLa细胞活力、侵袭和迁移能力;通过实时荧光定量聚合酶链反应(RT-qPCR)检测miR-802和PTCH1 mRNA的表达水平。通过生物信息学分析以及双荧光素酶报告基因系统验证了miR-802和PTCH1之间的靶关系。分别将NC mimic、miR-802 mimic转染至HeLa细胞中,命名为NC mimic组、miR-802 mimic组。将NC inhibitor、miR-802 inhibitor、Vector和pcDNA-PTCH1转染至10μg/mL丙泊酚处理的HeLa细胞中,分别命名为丙泊酚+NC inhibitor组、丙泊酚+miR-802 inhibitor组、丙泊酚+Vector组和丙泊酚+pcDNA-PTCH1组,检测细胞活力、侵袭和迁移能力。采用蛋白质印迹法检测PTCH1和Gli1蛋白表达水平。结果:与对照组比较,丙泊酚组HeLa细胞活力、侵袭和迁移能力均显著降低,差异均有统计学意义(P<0.05)。与NC mimic组比较,miR-802 mimic组HeLa细胞中的miR-802表达水平显著升高,细胞活力降低,侵袭和迁移能力降低,差异均有统计学意义(P<0.05)。与对照组比较,丙泊酚+NC inhibitor组HeLa细胞中miR-802表达水平显著升高;与丙泊酚+NC inhibitor组比较,丙泊酚+miR-802 inhibitor组HeLa细胞中miR-802表达水平显著降低,细胞活力显著增强,侵袭和迁移能力显著增强,上述差异均有统计学意义(P<0.05)。PTCH1被鉴定为miR-802的靶基因。与NC inhibitor组比较,miR-802 inhibitor组细胞中PTCH1 mRNA和蛋白水平显著升高,Gli1蛋白表达水平升高,差异均有统计学意义(P<0.05)。过表达PTCH1逆转了miR-802和丙泊酚诱导的对HeLa细胞活力、迁移和侵袭能力的抑制。结论:丙泊酚通过调节miR-802/PTCH1/SHH通路来抑制宫颈癌细胞的迁移和侵袭。OBJECTIVE: To probe into the effects of propofol on migration and invasion of cervical cancer cells through miR-802/PTCH1/SHH pathway. METHODS: Human cervical cancer cells HeLa cells were treated with different concentrations of propofol as the propofol group and dimethyl sulfoxide as the control group. MTT and Transwell were used to detect HeLa cell viability, invasion and migration ability. Real-time fluorescent quantitative polymerase chain reaction(RT-qPCR) was used to detect the expression level of miR-802 and PTCH1 mRNA. The target relationship between miR-802 and PTCH1 was verified by bioinformatics analysis and dual luciferase reporter gene system. NC mimic and miR-802 mimic were transfected into HeLa cells and named as NC mimic group and miR-802 mimic group. NC inhibitor, miR-802 inhibitor, Vector, and pcDNA-PTCH1 were transfected into HeLa cells treated with 10 μg/mL propofol and respectively named as propofol+NC inhibitor group, propofol+miR-802 inhibitor group, propofol+Vector group and propofol+pcDNA-PTCH1 group, cell viability, invasion and migration were detected. Western blotting was used to detect the protein expression level of PTCH1 and Gli1. RESULTS: Compared with the control group, the invasion and migration of HeLa cells in the propofol group decreased significantly, with statistically significant difference(P<0.05). Compared with the NC mimic group, the expression of miR-802 in HeLa cells in miR-802 mimic group increased significantly, and the cell viability, invasion and migration ability decreased, the differences were statistically significant(P<0.05). Compared with the control group, the expression of miR-802 in HeLa cells in the propofol+NC inhibitor group increased significantly;compared with the propofol+NC inhibitor group, the expression of miR-802 in HeLa cells in the propofol+miR-802 inhibitor group decreased significantly, the cell viability, invasion and migration ability increased, the differences were statistically significant(P<0.05). PTCH1 was identified as the target gene

关 键 词：丙泊酚 宫颈癌 miR-802 PTCH1 SHH 

分 类 号：R971.2[医药卫生—药品] R979.1[医药卫生—药学]