【摘要】：目的即刻早期反應(yīng)基因(immediate early response gene X-1,IEX-1)是一種凋亡易感基因,在多種人類惡性腫瘤中表達失調(diào),但在宮頸癌組織中的表達相關(guān)研究較少,檢測宮頸癌組織IEX-1的表達,了解其在宮頸癌發(fā)生發(fā)展中的作用,分析其與人乳頭瘤病毒(human papillomavirus,HPV)感染的關(guān)系。方法收集2011-01-01-2015-10-31鄭州大學第一附屬醫(yī)院病理科存檔蠟塊標本132份,采用免疫組化法檢測IEX-1在宮頸癌、宮頸上皮內(nèi)瘤變(cervical intraepithelial neoplasia,CIN)及良性宮頸病變組織中的表達情況,分析IEX-1的表達與宮頸癌臨床病理關(guān)系。采用RT-qPCR法及蛋白質(zhì)印跡法檢測Siha(HPV16+)、HeLa(HPV18+)和C-33a(HPV-)3種宮頸癌細胞株中IEX-1mRNA及蛋白的表達,siRNA法靶向沉默E6后Siha細胞E6、IEX-1的mRNA及蛋白表達,分析與HPV感染的關(guān)系。結(jié)果 IEX-1在宮頸良性病變、CIN組織和宮頸癌組織中的表達分別為72.4%、58.7%和21.1%,差異有統(tǒng)計學意義,χ~2=21.345,P=0.001。宮頸癌組織IEX-1的表達與患者年齡、病理類型、HPV感染型別及FIGO分期無關(guān),均P0.05。IEX-1的表達與宮頸癌分化程度、淋巴結(jié)轉(zhuǎn)移情況及宮旁浸潤密切相關(guān),均P0.05。C-33a細胞中IEX-1mRNA表達量為21.652±9.020,明顯高于Siha細胞的1(t=7.34,P0.001)和HeLa細胞的1.742±0.854(t=6.21,P0.001);IEX-1蛋白表達量為18.017±1.351,明顯高于Siha細胞的1.519±0.391(t=8.16,P0.001)和HeLa細胞的1.957±0.363(t=7.19,P0.001)。干擾組E6mRNA水平為0.218±0.106,低于陰性對照組的1.031±0.072(t=9.21,P0.001)和空白對照組(t=8.46,P0.001);干擾組E6蛋白表達量為0.184±0.274,低于陰性對照組的0.692±0.133(t=8.71,P0.001)和空白對照組的0.783±0.097(t=8.06,P0.001)。干擾組IEX-1mRNA水平為20.083±0.672,低于陰性對照組的1.193±0.408(t=9.04,P0.001)和空白對照組,t=7.56,P0.001;干擾組IEX-1蛋白表達量為0.672±0.135,低于陰性對照組的0.137±0.071(t=7.92,P0.001)和空白對照組的0.081±0.009(t=6.47,P0.001)。結(jié)論IEX-1在宮頸癌中表達降低,其表達水平與宮頸癌惡性程度呈負相關(guān),可能參與了宮頸癌的發(fā)生與發(fā)展。IEX-1在宮頸癌細胞中的表達與HPV感染有關(guān),但與感染的HPV型別無關(guān)。
[Abstract]:Objective immediate early response gene (immediate early response gene X-1nIEX-1) is a kind of apoptotic susceptible gene, which is misexpressed in various human malignant tumors. However, there are few studies on the expression of IEX-1 in cervical cancer tissues, and the expression of IEX-1 in cervical cancer tissues is detected. To understand the role of human papillomavirus (human papillomavirus,HPV) in the occurrence and development of cervical cancer and to analyze its relationship with human papillomavirus (human papillomavirus,HPV) infection. Methods 132 paraffin pieces were collected from Department of Pathology, Zhengzhou University from January 2011 to January 2015-31. The expression of IEX-1 in cervical carcinoma, cervical intraepithelial neoplasia (cervical intraepithelial neoplasia,CIN) and benign cervical lesions was detected by immunohistochemical method. To analyze the relationship between the expression of IEX-1 and the clinicopathology of cervical cancer. The expression of IEX-1mRNA and protein in three cervical cancer cell lines Siha (HPV16) HeLa (HPV18) and C-33a (HPV-) were detected by RT-qPCR and Western blotting. Results the expression of IEX-1 in cervical benign lesions cin and cervical carcinoma tissues were 72.4% and 21.1%, respectively. The difference was statistically significant. The expression of IEX-1 in cervical carcinoma was not related to age, pathological type and FIGO stage. The expression of P0.05.IEX-1 was closely related to the degree of cervical cancer differentiation, lymph node metastasis and periuterine invasion. The expression of IEX-1mRNA in P0.05.C-33a cells was 21.652 鹵9.020, which was significantly higher than that in Siha cells and HeLa cells (1.742 鹵0.854 and 18.017 鹵1.351, respectively), which was significantly higher than that in Siha cells (1.519 鹵0.391) and HeLa cells (1.957 鹵0.363). The E6mRNA level of the interference group was 0.218 鹵0.106, which was lower than that of the negative control group (1.031 鹵0.072) and the blank control group (t = 8.46), the expression of E6 protein in the interference group was 0.184 鹵0.274, which was lower than that in the negative control group (0.692 鹵0.133) and the blank control group (0.783 鹵0.097). The level of IEX-1mRNA in the interference group was 20.083 鹵0.672, which was lower than that in the negative control group (1.193 鹵0.408) and the blank control group (7.56 P 0.001), and the expression of IEX-1 protein in the interference group was 0.672 鹵0.135, which was lower than that in the negative control group (0.137 鹵0.071) and the blank control group (0.081 鹵0.009). Conclusion the expression of IEX-1 in cervical cancer is decreased, and its expression level is negatively correlated with the degree of malignancy of cervical cancer. The expression of IEX-1 may be involved in the occurrence and development of cervical cancer. The expression of IEX-1 in cervical cancer cells is related to HPV infection, but not to the type of HPV infection.
【作者單位】： 鄭州大學第一附屬醫(yī)院婦科·河南省高等學校臨床醫(yī)學重點學科開放實驗室;
【基金】：河南省科技廳重點攻關(guān)項目(122102310631)
【分類號】：R737.33

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