miR-128调控AK2蛋白的表达通过STAT3信号通路对宫颈癌细胞生物学行为的影响  

Effect of miR-128 and AK2 on biological behavior of cervical cancer cells by STAT3 signaling pathway

作　　者：杨林青[1] 孙颖川 YANG Lin-Qing;SUN Ying-Chuan(The University of Xuchang ,Xuchang 461000, China)

机构地区：[1]许昌学院,许昌461000

出　　处：《中国免疫学杂志》2017年第12期1783-1788,共6页Chinese Journal of Immunology

基　　金：河南省医学科技攻关计划项目(201203068)

摘　　要：目的:探讨miR-128调控AK2蛋白的表达通过STAT3信号通路对宫颈癌细胞生物学行为的影响其机制。方法:q PCR和Western blot检测宫颈癌组织和细胞中AK2和miR-128的表达情况;双荧光素酶实验检测miR-128和AK2之间的相互作用;CCK-8增殖试验检测miR-128对宫颈癌细胞增值能力的影响;裸鼠体内成瘤试验检测miR-128对宫颈癌细胞成瘤能力的影响;Western blot检测miR-128对STAT3信号通路蛋白水平的影响;Western blot检测过表达AK2蛋白逆转miR-128对p-STAT3的抑制水平。结果:在宫颈癌组织中AK2表达水平相比正常宫颈组织中高,miR-128在宫颈癌C33a细胞株中表达水平较低;双荧光素酶试验证实miR-128可以直接靶向调控AK2的表达;CCK-8增殖实验表明miR-128可以抑制宫颈癌细胞株的增殖能力;体内成瘤实验表明miR-128的增高可以抑制宫颈癌细胞成瘤能力[体积(3.05±0.35)cm3vs(0.86±0.11)cm3,P=0.031;(3.26±0.39)g vs(0.89±0.15)g,P=0.016];Western blot实验表明miR-128可以抑制p-STAT的激活[(42.12±6.28)%vs(91.25±9.29)%,P<0.05],而AK2的过表达又可以逆转miR-128对p-STAT的抑制作用。结论:miR-128靶向调控AK2的表达进而通过STAT3通路的激活调控宫颈癌细胞的生物学行为。Objective：To investigate the mechanism of miR-128 on the expression of AK2 protein through the STAT3 signaling pathway on the biological behavior of cervical cancer cells .Methods： The expression of AK2 and miR-128 in cervical cancer tissues and cells was detected by qPCR and Western blot .Double luciferase assay was used to detect the interaction between miR-128 and AK2.CCK-8 proliferation assay was used to detect the effect of miR-128 on the enhancement of cervical cancer cells .The effect of miR-128 on the tumorigenesis of cervical cancer cells was exa mined by tumor formation test in nude mice .Western blot was used to detect the effect of miR-128 on STAT3 signal pathway protein level .Western blot was used to detect the inhibitory effect of miR-128 on p-STAT3 by overexpressing AK2 protein.Results：The expression level of AK2 in cervical cancer tissues was higher than that in normal cervical tissues,and the expression level of miR-128 in cervical cancer C33a cells was lower.Double luciferase assay confirmed that miR-128 could directly target the expression of AK 2.CCK-8 proliferation test showed that miR-128 could inhibit the proliferation of cervical cancer cell lines .In vivo tumorigenesis test showed that the increase of miR-128 could inhibit the tumorigenesis ability of cervical cancer cells[volume（3.05±0.35）cm3 vs （0.86±0.11）cm3,P=0.031;weight（3.26±0.39）g vs （0.89±0.15）g,P=0.016 ];Western blot showed that miR-128 could inhibit the activation of p-STAT [ （ 42.12 ±6.28 ）% vs （ 91.25 ±9.29 ）%, P〈0.05 ] ,while the overexpression of AK 2 could reverse the inhibitory effect of miR-128 on p-STAT.Conclusion： miR-128 is used to regulate the expression of AK 2 and regulate the biological behavior of cervical cancer cells through the activation of STAT 3 pathway.

关 键 词：miR-128 宫颈癌 AK2 CCK-8 

分 类 号：R737.33[医药卫生—肿瘤]