發(fā)布時(shí)間：2018-11-08 13:58

【摘要】：本實(shí)驗(yàn)旨在探討EZH2基因在宮頸癌中表達(dá)情況、對(duì)宮頸癌細(xì)胞增殖能力的影響及其機(jī)制。通過(guò)免疫組織化學(xué)法檢測(cè)20例正常宮頸、20例CIN及60例宮頸鱗癌組織中EZH2蛋白表達(dá);采用RT-PCR法檢測(cè)4個(gè)宮頸癌細(xì)胞株中EZH2 m RNA的表達(dá)。通過(guò)Lipofectamin2000介導(dǎo)EZH2 si RNA瞬時(shí)轉(zhuǎn)染C33A細(xì)胞株,Western blotting法檢測(cè)si RNA的干擾效率。MTT、流式細(xì)胞儀檢測(cè)EZH2沉默后細(xì)胞增殖及細(xì)胞周期變化;Western blotting檢測(cè)p21表達(dá)變化。結(jié)果顯示,正常宮頸組織、CIN、宮頸鱗癌中EZH2陽(yáng)性表達(dá)率分別為15%、30%、76.7%,呈逐級(jí)升高趨勢(shì),后者與前兩者見(jiàn)差異有統(tǒng)計(jì)學(xué)意義(p0.01);EZH2的表達(dá)同宮頸鱗癌細(xì)胞分化程度、間質(zhì)浸潤(rùn)深度及淋巴結(jié)轉(zhuǎn)移顯著相關(guān)(p0.05)。EZH2 m RNA在宮頸癌細(xì)胞株He La、Si Ha、C33A及Caski中均表達(dá),其中C33A細(xì)胞中表達(dá)最高。si RNA沉默EZH2基因明顯抑制宮頸癌C33A細(xì)胞增殖,阻滯細(xì)胞周期于G1期;Western blotting檢測(cè)p21表達(dá)上調(diào)。由此得出結(jié)論,EZH2基因在宮頸癌中高表達(dá);沉默EZH2基因可通過(guò)上調(diào)p21蛋白的表達(dá)阻滯細(xì)胞周期于G1期、抑制宮頸癌細(xì)胞的增殖。
[Abstract]:The aim of this study was to investigate the expression of EZH2 gene in cervical cancer and its mechanism. The expression of EZH2 protein in 20 cases of normal cervix, 20 cases of CIN and 60 cases of cervical squamous cell carcinoma were detected by immunohistochemical method, and the expression of EZH2 m RNA in 4 cervical cancer cell lines was detected by RT-PCR method. The interference efficiency of si RNA was detected by Lipofectamin2000 mediated EZH2 si RNA transient transfection of C33A cell line, Western blotting, and the expression of p21 was detected by MTT, flow cytometry after EZH2 silencing and; Western blotting expression. The results showed that the positive expression rate of EZH2 in normal cervical tissues and CIN, cervical squamous cell carcinoma was 1530 and 76.7respectively, which showed a gradual increase trend, and the difference between the latter and the former was statistically significant (p0.01). The expression of EZH2 was significantly correlated with the differentiation of cervical squamous cell carcinoma, the depth of interstitial infiltration and lymph node metastasis (p0.05). EZH2 m RNA was expressed in both He La,Si Ha,C33A and Caski cells. The highest expression of. Si RNA silencing EZH2 gene in C33A cells significantly inhibited the proliferation of cervical cancer C33A cells and blocked the cell cycle in G1 phase. The expression of p21 was up-regulated by Western blotting. It is concluded that EZH2 gene is highly expressed in cervical cancer and silencing EZH2 gene can inhibit the proliferation of cervical cancer cells by up-regulating the expression of p21 protein to block cell cycle in G1 phase.
【作者單位】： 西安交通大學(xué)第一附屬醫(yī)院婦產(chǎn)科;
【基金】：陜西省國(guó)際科技合作與交流計(jì)劃項(xiàng)目基金(2013KW-27-01) 中央高�；究蒲袠I(yè)務(wù)費(fèi)專項(xiàng)資金(XJJ2015-094)共同資助
【分類號(hào)】：R737.33

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本文編號(hào)：2318694