宫颈癌组织IEX-1表达及其与HPV感染相关性研究

发布时间：2018-10-05 15:31

【摘要】：目的即刻早期反应基因(immediate early response gene X-1,IEX-1)是一种凋亡易感基因,在多种人类恶性肿瘤中表达失调,但在宫颈癌组织中的表达相关研究较少,检测宫颈癌组织IEX-1的表达,了解其在宫颈癌发生发展中的作用,分析其与人乳头瘤病毒(human papillomavirus,HPV)感染的关系。方法收集2011-01-01-2015-10-31郑州大学第一附属医院病理科存档蜡块标本132份,采用免疫组化法检测IEX-1在宫颈癌、宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)及良性宫颈病变组织中的表达情况,分析IEX-1的表达与宫颈癌临床病理关系。采用RT-qPCR法及蛋白质印迹法检测Siha(HPV16+)、HeLa(HPV18+)和C-33a(HPV-)3种宫颈癌细胞株中IEX-1mRNA及蛋白的表达,siRNA法靶向沉默E6后Siha细胞E6、IEX-1的mRNA及蛋白表达,分析与HPV感染的关系。结果 IEX-1在宫颈良性病变、CIN组织和宫颈癌组织中的表达分别为72.4%、58.7%和21.1%,差异有统计学意义,χ~2=21.345,P=0.001。宫颈癌组织IEX-1的表达与患者年龄、病理类型、HPV感染型别及FIGO分期无关,均P0.05。IEX-1的表达与宫颈癌分化程度、淋巴结转移情况及宫旁浸润密切相关,均P0.05。C-33a细胞中IEX-1mRNA表达量为21.652±9.020,明显高于Siha细胞的1(t=7.34,P0.001)和HeLa细胞的1.742±0.854(t=6.21,P0.001);IEX-1蛋白表达量为18.017±1.351,明显高于Siha细胞的1.519±0.391(t=8.16,P0.001)和HeLa细胞的1.957±0.363(t=7.19,P0.001)。干扰组E6mRNA水平为0.218±0.106,低于阴性对照组的1.031±0.072(t=9.21,P0.001)和空白对照组(t=8.46,P0.001);干扰组E6蛋白表达量为0.184±0.274,低于阴性对照组的0.692±0.133(t=8.71,P0.001)和空白对照组的0.783±0.097(t=8.06,P0.001)。干扰组IEX-1mRNA水平为20.083±0.672,低于阴性对照组的1.193±0.408(t=9.04,P0.001)和空白对照组,t=7.56,P0.001;干扰组IEX-1蛋白表达量为0.672±0.135,低于阴性对照组的0.137±0.071(t=7.92,P0.001)和空白对照组的0.081±0.009(t=6.47,P0.001)。结论IEX-1在宫颈癌中表达降低,其表达水平与宫颈癌恶性程度呈负相关,可能参与了宫颈癌的发生与发展。IEX-1在宫颈癌细胞中的表达与HPV感染有关,但与感染的HPV型别无关。
[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 卤9.020, which was significantly higher than that in Siha cells and HeLa cells (1.742 卤0.854 and 18.017 卤1.351, respectively), which was significantly higher than that in Siha cells (1.519 卤0.391) and HeLa cells (1.957 卤0.363). The E6mRNA level of the interference group was 0.218 卤0.106, which was lower than that of the negative control group (1.031 卤0.072) and the blank control group (t = 8.46), the expression of E6 protein in the interference group was 0.184 卤0.274, which was lower than that in the negative control group (0.692 卤0.133) and the blank control group (0.783 卤0.097). The level of IEX-1mRNA in the interference group was 20.083 卤0.672, which was lower than that in the negative control group (1.193 卤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 卤0.135, which was lower than that in the negative control group (0.137 卤0.071) and the blank control group (0.081 卤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.
【作者单位】： 郑州大学第一附属医院妇科·河南省高等学校临床医学重点学科开放实验室;
【基金】：河南省科技厅重点攻关项目(122102310631)
【分类号】：R737.33

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