E2F2诱导对非小细胞肺癌细胞增殖及其不良预后的相关性研究

发布时间：2018-05-14 10:45

本文选题：E2F2 + 非小细胞肺癌　；参考：《南昌大学》2016年博士论文

【摘要】：非小细胞肺癌是全球最常见的癌症之一,在美国和中国,肺癌都是致死率最高的癌症。E2F转录调节因子参与了大量的生命活动,包括细胞周期、细胞凋亡以及DNA损伤应答。本研究对E2F2是否参与非小细胞肺癌生长、增殖,及其与非小细胞肺癌预后的关系,HPV感染与NSCLC中E2F2、Survivin蛋白表达的相关性进行了探讨。方法:1、采集86例经病理或细胞学确诊的非小细胞肺癌患者的组织通过免疫组化的方式分析E2F2蛋白的表达水平,并统计分析E2F2与临床特征和不良预后的相关性。对其中8对非小细胞肺癌患者手术切除后肺癌组织及癌旁组织,通过运用Western Blot方法检测肺癌组织及癌旁组织研究E2F2在细胞中表达的量。2、构建E2F2 shRNA和杂序shRNA,以慢病毒转染H1299和A549非小细胞肺癌细胞系,用MTT、免疫组化和菌落形成实验来研究转染后E2F2表达的改变,以及E2F2对细胞增殖和生长的影响。3、选取86例非小细胞肺癌和26例良性病变肺组织,每例标本均分成两组:一组应用PCR的方式用人乳头状瘤病毒(HPV)的16E6和18E7这两种物质进行特异性的引物来探究上述组织中是否有对应的物质存在;另一组用免疫组化的方式检测E2F2和Survivin蛋白在良性组织中和非小细胞肺癌的表达情况,采用统计学的方法来分析其数据。结果:1.通过免疫组化的方式分析了86例非小细胞肺癌的结果表明,86例组织中,有54例E2F2的表达量相比癌旁正常的组织有显著性的提高。用western blot检测了对其中8对非小细胞肺癌患者的癌组织及其癌旁组织,结果表明在8组中,有5组的结果显示E2F2的表达量在癌组织中要显著的高于癌旁正常组织。,western blot结果在8对非小细胞肺癌癌组织及癌旁正常组织中,有5组的结果显示E2F2的表达量在癌组织中要显著的高于癌旁正常组织。2.进一步的研究表明E2F2的表达量与临床分期(P=0.039)和肿瘤大小(P=0.045)存在这一定的关系。此外,Kaplan-Meier分析表明高的高表达与总生存期(P=0.045)显著性相关,但是与无病生存期(P=0.288)不相关。3.结果显示抑制E2F2的表达能够降低非小细胞肺癌细胞的活性以及集落形成的能力。4.非小细胞肺癌组中HPV16和18型DNA检出率为43.00%(37/86),对于良性肺病组的7.69%(2/26)发生的明显的偏高,两者间存着显著性差异。5.E2F2蛋白在非小细胞肺癌组织中的表达率62.80%(54/86),对比肺良性病变组的23.08%(6/26),表达率要明显的偏高,两者间也存在着显著性差异。其中E2F2阳性表达中的高表达率为64.81%(35/54)。且Survivin蛋白在非小细胞肺癌中表达的阳性率为61.63%(53/86),Survivin蛋白的表达在肺良性病变组中没有检测到。6.在86例非小细胞肺癌中,E2F2和Survivin的阳性表达数分别为54和53例,其中两者均阳性表达的数目为33例,两者均为阴性表达的例数为17例,通过计数资料的相关分析得出在非小细胞肺癌中这两者的表达量呈正相关。7.E2F2和Survivin蛋白在非小细胞肺癌中HPV阳性组中的阳性表达率分别为86.50%(32/37)、78.40%(29/37),均高于HPV阴性组中的38.8%(19/49)、49.0%(24/49),两者间存在着显著性差异。结论:本实验显示E2F2高表达者有较低的总生存,提示E2F2可能是肺癌预后不良因素之一。单变量多变量分析结果显示E2F2高表达是总生存的一个相对独立的预后因素。抑制E2F2的表达能够降低非小细胞肺癌细胞的活性以及集落形成的能力,提示E2F2参与了非小细胞肺癌细胞增殖。非小细胞肺癌中HPV的检出率显著高于良性肺病变,HPV可能是非小细胞肺癌发生发展的重要病因之一。HPV可能是通过提高E2F2和Survivin的表达量来促使非小细胞肺癌的生长增殖。
[Abstract]:Non-small cell lung cancer (non-small cell lung cancer) is one of the most common cancers in the world. In both the United States and China, lung cancer is the most fatal cancer.E2F transcriptional regulator involved in a large number of life activities, including cell cycle, cell apoptosis and DNA damage response. This study is a study of whether E2F2 is involved in the growth, proliferation, and non small cell lung of non-small cell lung cancer. The relationship between cancer prognosis, HPV infection and the correlation of E2F2 and Survivin protein expression in NSCLC. Methods: 1, the expression of E2F2 protein was analyzed by immunohistochemical method in 86 cases of non small cell lung cancer diagnosed by pathological or cytological diagnosis, and the correlation between E2F2 and clinical characteristics and poor prognosis was statistically analyzed. 8 of the cases of non small cell lung cancer after resection of lung cancer tissue and para cancer tissue, by using Western Blot method to detect lung cancer tissue and para cancer tissue to study the expression of E2F2 in the cell.2, E2F2 shRNA and miscellaneous shRNA, lentivirus transfection H1299 and A549 non-small cell lung cancer cell lines, MTT, immunohistochemical and colony shape The changes in the expression of E2F2 after transfection, and the effect of E2F2 on cell proliferation and growth,.3, 86 cases of non small cell lung cancer and 26 benign lung tissues were selected, each of which was divided into two groups: a group of two specific primers used to explore the 16E6 and 18E7 of human papillomavirus (HPV) using PCR. Whether there was a corresponding substance in the tissue, another group was used to detect the expression of E2F2 and Survivin protein in benign and non small cell lung cancer by immunohistochemistry. Statistical analysis was used to analyze the data. Results: 1. the results of 86 cases of non small cell lung cancer were analyzed by immunohistochemical method, and 86 cases of non small cell lung cancer were analyzed. In 54 cases, the expression of E2F2 was significantly higher than that of the normal tissue adjacent to the cancer. Western blot was used to detect the cancer tissue and paracancerous tissue in 8 of the patients with non-small cell lung cancer. The results showed that in the 8 groups, the results of 5 groups showed that the expression of E2F2 was significantly higher than that in the normal tissue adjacent to the cancer tissue. The Western blot result. In 8 pairs of non-small cell lung cancer tissues and adjacent normal tissues, the results of 5 groups showed that the expression of E2F2 was significantly higher in the cancer tissue than in the normal tissue adjacent to the cancer. The further study of the.2. showed that the expression of E2F2 was related to the existence of the clinical stage (P=0.039) and the tumor size (P=0.045). Furthermore, the Kaplan-Meier analysis showed that the expression was high. The high expression was significantly related to the total survival time (P=0.045), but the unrelated.3. results with the disease-free survival (P=0.288) showed that inhibition of the expression of E2F2 could reduce the activity of non small cell lung cancer cells and the ability of colony formation. The HPV16 and type 18 DNA detection rate in non small cell lung cancer group was 43% (37/86), and 7.69% for the benign lung disease group. 2/26) was significantly higher, and there was a significant difference in the expression of.5.E2F2 protein in non small cell lung cancer tissue, 62.80% (54/86), compared with 23.08% (6/26) in the benign lung disease group, the expression rate was significantly higher, and there was a significant difference between the two. The high expression rate in the positive expression of E2F2 was 64.81% (35/54). And Survi The positive rate of VIN protein expression in non-small cell lung cancer was 61.63% (53/86). The expression of Survivin protein was not detected in 86 cases of non small cell lung cancer in the benign lung disease group. The positive expressions of E2F2 and Survivin were 54 and 53, respectively, of which both were positive in 33 cases, and the number of both negative expressions was 17. By the correlation analysis of counting data, the positive expression of both.7.E2F2 and Survivin protein in non small cell lung cancer was 86.50% (32/37) and 78.40% (29/37) in non small cell lung cancer, respectively, which were higher than 38.8% (19/49) and 49% (24/49) in the negative group of HPV (24/49), and there was a significant difference between them. Differences. Conclusion: this experiment shows that the high expression of E2F2 has a lower total survival, suggesting that E2F2 may be one of the adverse factors for the prognosis of lung cancer. The results of single variable multivariate analysis show that high expression of E2F2 is a relatively independent prognostic factor for total survival. Inhibition of the expression of E2F2 can reduce the activity of non small cell lung cancer cells and the formation of colony formation. The ability to suggest that E2F2 participates in non-small cell lung cancer cell proliferation. The detection rate of HPV in non-small cell lung cancer is significantly higher than that of benign lung disease. HPV may be one of the important causes of the development of non-small cell lung cancer (.HPV) may be to promote the growth and proliferation of non-small cell lung cancer by increasing the expression of E2F2 and Survivin.

【学位授予单位】：南昌大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R734.2

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