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

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:qPCR 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)cm3 vs (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。

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

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 AK2.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 cellsvolume(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 AK2 could reverse the inhibitory effect of miR-128 on p-STAT.Conclusion: miR-128 is used to regulate the expression of AK2 and regulate the biological behavior of cervical cancer cells through the activation of STAT3 pathway.