發(fā)布時(shí)間：2018-12-10 21:55

【摘要】：目的:探討miR-128調(diào)控AK2蛋白的表達(dá)通過STAT3信號(hào)通路對(duì)宮頸癌細(xì)胞生物學(xué)行為的影響其機(jī)制。方法:q PCR和Western blot檢測(cè)宮頸癌組織和細(xì)胞中AK2和miR-128的表達(dá)情況;雙熒光素酶實(shí)驗(yàn)檢測(cè)miR-128和AK2之間的相互作用;CCK-8增殖試驗(yàn)檢測(cè)miR-128對(duì)宮頸癌細(xì)胞增值能力的影響;裸鼠體內(nèi)成瘤試驗(yàn)檢測(cè)miR-128對(duì)宮頸癌細(xì)胞成瘤能力的影響;Western blot檢測(cè)miR-128對(duì)STAT3信號(hào)通路蛋白水平的影響;Western blot檢測(cè)過表達(dá)AK2蛋白逆轉(zhuǎn)miR-128對(duì)p-STAT3的抑制水平。結(jié)果:在宮頸癌組織中AK2表達(dá)水平相比正常宮頸組織中高,miR-128在宮頸癌C33a細(xì)胞株中表達(dá)水平較低;雙熒光素酶試驗(yàn)證實(shí)miR-128可以直接靶向調(diào)控AK2的表達(dá);CCK-8增殖實(shí)驗(yàn)表明miR-128可以抑制宮頸癌細(xì)胞株的增殖能力;體內(nèi)成瘤實(shí)驗(yàn)表明miR-128的增高可以抑制宮頸癌細(xì)胞成瘤能力[體積(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實(shí)驗(yàn)表明miR-128可以抑制p-STAT的激活[(42.12±6.28)%vs(91.25±9.29)%,P0.05],而AK2的過表達(dá)又可以逆轉(zhuǎn)miR-128對(duì)p-STAT的抑制作用。結(jié)論:miR-128靶向調(diào)控AK2的表達(dá)進(jìn)而通過STAT3通路的激活調(diào)控宮頸癌細(xì)胞的生物學(xué)行為。
[Abstract]:Aim: to investigate the effect of miR-128 on the biological behavior of cervical cancer cells through STAT3 signaling pathway. Methods: q PCR and Western blot were used to detect the expression of AK2 and miR-128 in cervical cancer tissues and cells, and double luciferase assay was used to detect the interaction between miR-128 and AK2. CCK-8 proliferation test was used to detect the effect of miR-128 on the proliferation of cervical cancer cells and in vivo tumorigenesis test in nude mice to detect the effect of miR-128 on the tumorigenic ability of cervical cancer cells.; Western blot was used to detect the effect of miR-128 on the level of STAT3 signal pathway protein. Overexpression of AK2 protein was detected by Western blot to reverse the inhibition of p-STAT3 by miR-128. Results: the expression of AK2 in cervical carcinoma was higher than that in normal cervix, and the expression of miR-128 was lower in cervical cancer C33a cell line, and double luciferase assay showed that miR-128 could directly target the expression of AK2. CCK-8 proliferation assay showed that miR-128 could inhibit the proliferation of cervical cancer cell line. In vivo tumorigenesis experiment showed that the increase of miR-128 could inhibit the tumorigenic ability of cervical cancer cells [volume (3.05 鹵0.35) cm3vs (0.86 鹵0.11) cm3,P=0.031; (3.26 鹵0.39) g vs (0.89 鹵0.15) g P0. 016]; Western blot assay showed that miR-128 could inhibit the activation of p-STAT [(42.12 鹵6.28)% vs (91.25 鹵9.29)%, P0.05], while the overexpression of AK2 could reverse the inhibitory effect of miR-128 on p-STAT. Conclusion: miR-128 can regulate the expression of AK2 and regulate the biological behavior of cervical cancer cells through the activation of STAT3 pathway.
【作者單位】： 許昌學(xué)院;
【基金】：河南省醫(yī)學(xué)科技攻關(guān)計(jì)劃項(xiàng)目(201203068)
【分類號(hào)】：R737.33

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