摘要
目的探讨过表达miR-411对宫颈癌细胞增殖、侵袭和凋亡的影响及机制。方法以正常宫颈上皮永生化细胞End1/E6E7为对照细胞,qRT-PCR检测宫颈癌Caski、SiHa和C-33A细胞miR-411表达。MTT法、Transwell小室及流式细胞术检测转染miR-411 mimics 48 h的SiHa细胞增殖、侵袭能力和凋亡率。通过双荧光素酶报告基因实验及miR-411 mimics/inhibitor对PIK3R3表达影响验证miR-411和PIK3R3靶向关系。将PIK3R3 siRNA及miR-411 mimics+PIK3R3过表达载体转染至SiHa细胞,转染48 h,检测细胞的增殖、侵袭能力和凋亡率。Western blotting检测转染miR-411 mimics 48 h的SiHa细胞PI3K、p-AKT、PCNA、E-cadherin和cleaved caspase3表达。结果miR-411在宫颈癌细胞表达明显降低,差异有统计学意义(P<0.05)。过表达miR-411可明显抑制SiHa细胞增殖和侵袭能力,促进细胞凋亡(P<0.05)。miR-411可靶向负调控PIK3R3表达。抑制PIK3R3表达可明显降低SiHa细胞增殖和侵袭能力,促进细胞凋亡(P<0.05)。过表达PIK3R3可逆转miR-411对SiHa细胞增殖、侵袭和凋亡的影响。过表达miR-411可明显抑制PI3K、p-AKT和PCNA表达,促进E-cadherin和cleaved caspase3表达(P<0.05)。结论miR-411可通过靶向PIK3R3抑制PI3K/AKT信号通路,从而影响宫颈癌细胞增殖、侵袭和凋亡。
Objective To investigate the effects of miR-411 overexpression on the proliferation,invasion and apoptosis of cervical cancer cells.Methods Taking End1/E6E7 as control group,the expression levels of miR-411 in CaSki,SiHa and C-33A cells were detected by qRT-PCR.MTT assay,Transwell chamber and flow cytometry were used to detect the proliferation,invasion and apoptosis rate of SiHa cells which were transfected with miR-411 mimics for 48h.The targeted relationship between miR-411 and PIK3R3 was verified by double luciferase reporter gene assay and the effect of miR-411 mimics/inhibitor on PIK3R3 expression.PIK3R3 siRNA and miR-411 mimics+PIK3R3 overexpressed vector were transfected into SiHa cells.The cells were transfected for 48 hours,the proliferation,invasion and apoptosis rate of the cells were detected.Western blotting was used to detect the expression of PI3K,p-AKT,PCNA,E-cadhenin and cleaved caspase-3 in SiHa cells transfected with miR-411 mimics for 48h.Results The expression levels of miR-411 in cervical cancer cells were significantly decreased(P<0.05).MiR-411 overexpression could significantly inhibit the proliferation and invasion of SiHa cells and promote apoptosis(P<0.05).MiR-411 could negatively targetedly regulate PIK3R3 expression.The inhibition of PIK3R3 expression could significantly reduce the proliferation and invasion of SiHa cells and promote apoptosis(P<0.05).Overexpression of PIK3R3 could reverse the effects of miR-411 on proliferation,invasion and apoptosis of SiHa cells.Moreover overexpression of miR-411 could significantly inhibit the expression of PI3K,p-AKT and PCNA,and promoted the expressions of E-cadherin and cleaved caspase-3(P<0.05).Conclusion MiR-411 can affect the proliferation,invasion and apoptosis of cervical cancer cells by targetedly regulating PIK3R3 to inhibit PI3K/AKT signaling pathway.
作者
唐亮
阴敏
张海
肖燕
马莹莹
TANG Liang;YIN Min;ZHANG Hai(Department of Gynecology,Panzhihua Hospital for Maternal and Child Health,Sichuan,Panzhihua 617000,China)
出处
《河北医药》
CAS
2022年第8期1130-1135,共6页
Hebei Medical Journal
基金
四川省卫生计划项目(编号:16PJ237)。
