BCL11A在宫颈癌组织中的表达及其在宫颈癌细胞中的作用及影响

发布时间：2018-05-16 18:36

本文选题：BCL11A + 淋巴结受累　；参考：《重庆医科大学》2017年硕士论文

【摘要】：宫颈癌是一种女性常见肿瘤,对女性身心健康造成了极大危害,近年来其发病率及死亡率在全球范围内呈上升趋势。虽然人类在宫颈癌HPV疫苗的研发中已有重大突破,但是全球范围内,宫颈癌依然是第四大癌症,其死亡率仅次于乳腺癌。B细胞淋巴瘤11A(B-cell CLL/lymphoma 11A,BCL11A),其同源基因Evi9(ecotropic viral integration site 9)最初发现于骨髓性白血病BXH2小鼠中,作为逆转录病毒的插入位点,并与BCL6蛋白相互作用[1]。BCL11A在非实体瘤如白血病、淋巴瘤等造血与淋巴组织肿瘤中发挥重要作用[2,6]。最近有研究发现,BCL11A在实体瘤如三阴性乳腺癌及非小细胞肺癌中也发挥着重要作用[3,4],但是BCL11A在宫颈癌中的作用尚未有报道,本研究将主要通过组织及细胞水平研究BCL11A在宫颈癌中的作用及对其机制进行初步探索。第一部分bcl11a在宫颈癌组织中的表达及其临床意义目的:研究bcl11a在宫颈癌组织及癌旁正常组织中的表达情况,并分析其差异表达与临床病理因素之间的关系及临床意义。方法:结合tcga数据库,利用生物信息学分析bcl11a在不同癌症组织及相应正常组织中的表达,并分析bcl11a的表达与宫颈癌患者预后的关系。收集62例宫颈癌组织及相对应的癌旁正常组织,制作成石蜡切片,并通过免疫组织化学法检测bcl11a蛋白在宫颈癌组织和癌旁正常组织中的表达,并结合患者资料,分析bcl11a的表达与临床病理因素之间的关系。结果:生物信息学分析得知bcl11a在宫颈癌及癌旁正常组织中存在差异;在收集的62例宫颈癌组织及癌旁正常组织中均检测到了bcl11a的表达,其中有50例宫颈癌组织检测到bcl11a的高表达(80.6%);有13例癌旁正常组织中检测到bcl11a的高表达(21%)。bcl11a蛋白在宫颈癌组织及癌旁正常组织中的表达存在差异(p0.01);bcl11a蛋白的表达与宫颈癌患者的年龄(p=0.436),肿瘤大小(p=0.734)以及淋巴结转移数目(p=0.439)等无关;与未出现淋巴结转移(n0)的宫颈癌患者相比,在出现淋巴结转移(n1-3)的宫颈癌患者中bcl11a蛋白的表达较高(p0.05);bcl11a表达较高的宫颈癌患者的总生存期要优于bcl11a表达较低的宫颈癌患者。在宫颈癌组织中,bcl11a的表达主要定位于细胞核,在宫颈癌正常组织中,bcl11a的表达主要定位于细胞质中。结论:bcl11a在宫颈癌组织中高表达,在癌旁正常组织中表达较低;bcl11a的表达与宫颈癌患者的淋巴结受累情况相关,即出现淋巴结转移的患者bcl11a的表达较高;在疾病发展的早期,bcl11a表达高的宫颈癌患者的总生存期要优于bcl11a表达低的宫颈癌患者,但并不能作为独立的宫颈癌患者总生存期的判定指标;在宫颈癌组织中bcl11a的表达主要定位于在细胞核中,而在癌旁正常组织中,bcl11a的表达主要定位于细胞质中。第二部分干扰BCL11A基因的表达对宫颈癌细胞相关生物学特性的影响目的:研究BCL11A基因对宫颈癌细胞周期、凋亡以及增殖能力的影响方法:设计并合成干扰BCL11A基因的si-RNA序列,筛选出干扰效果最佳序列;通过细胞转染靶向沉默BCL11A基因在宫颈癌C33A细胞中的表达,通过流式细胞技术检测沉默BCL11A后,C33A细胞周期及凋亡的变化情况;使用CCK-8试剂盒检测C33A细胞增殖能力的变化。结果:BCL11A在宫颈癌C33A细胞中表达较高,在Hela,Siha和Ca Ski细胞中表达较低;与siRNA2和siRNA3相比,siRNA1的干扰效率最好;与blank control组(6.56±0.59%)和NC组(7.85±0.47%)相比,siRNA-BCL11A组(28.45±4.07%)的细胞凋亡比率明显升高,且具有统计学意义(P0.05),但是三组的细胞周期变化及增殖能力无明显差异。结论:BCL11A在宫颈癌细胞中的表达水平可能与宫颈癌细胞的类型有关,在C33A细胞中沉默BCL11A基因的表达对C33A细胞的细胞周期及增殖能力没有影响,但是能够促进C33A细胞的凋亡。
[Abstract]:Cervical cancer is a common female tumor, which has caused great harm to the physical and mental health of women. In recent years, the incidence and mortality of the cervical cancer are rising worldwide. Although human beings have made great breakthroughs in the research and development of HPV vaccine for cervical cancer, cervical cancer is the fourth major cancer in the world, and the death rate is second to.B of breast cancer. Cell lymphoma 11A (B-cell CLL/lymphoma 11A, BCL11A), and its homologous gene Evi9 (ecotropic viral integration site 9) originally found in BXH2 mice with myeloid leukemia, as a retrovirus insertion site, and interacting with the BCL6 protein in non solid tumors such as leukemia, lymphoma and other hematopoietic and lymphoid tissue tumors. [2,6]. has recently shown that BCL11A also plays an important role in solid tumor such as three negative breast cancer and non-small cell lung cancer, [3,4], but the role of BCL11A in cervical cancer has not been reported. This study will mainly study the role and mechanism of BCL11A in cervical cancer by tissue and cell level. To explore the expression of Bcl11A in cervical cancer tissue and its clinical significance: To study the expression of Bcl11A in cervical cancer tissues and normal tissues adjacent to cancer, and to analyze the relationship and clinical significance between the differential expression and the clinicopathological factors. Methods: combining the TCGA data base and using bioinformatics to analyze Bcl11A in different carcinomas. The expression of Bcl11A and the prognosis of cervical cancer patients were analyzed. 62 cases of cervical cancer tissues and corresponding normal paracancerous tissues were collected, and paraffin sections were made, and the expression of Bcl11A protein in cervical cancer tissues and normal tissues was detected by immunohistochemistry and combined with the patients. Data, analysis of the relationship between the expression of Bcl11A and the clinicopathological factors. Results: bioinformatics analysis showed that Bcl11A was different in the normal tissues of cervical cancer and adjacent to cancer, and the expression of Bcl11A was detected in 62 cases of cervical cancer tissues and normal tissues adjacent to cancer, of which 50 cases of cervical cancer detected the high expression of Bcl11A (8 0.6%): the expression of high expression of Bcl11A (21%).Bcl11a protein in cervical cancer tissues and adjacent normal tissues was found in 13 normal para cancerous tissues (P0.01), and the expression of Bcl11A protein was not related to the age of cervical cancer (p=0.436), the size of the tumor (p=0.734) and the number of lymph node metastases (p=0.439), and no lymph nodes were found. The expression of Bcl11A protein in cervical cancer patients with metastatic (n1-3) metastasis (N0) was higher (P0.05) than that of cervical cancer patients with lymph node metastasis (P0.05); the total survival time of the patients with higher Bcl11A expression was better than that of the lower Bcl11A. In cervical cancer, the expression of Bcl11A was mainly located in the nucleus and in the normal cervical cancer. In the tissue, the expression of Bcl11A is mainly located in the cytoplasm. Conclusion: Bcl11A is highly expressed in cervical cancer tissues and low in normal tissues adjacent to cancer; the expression of Bcl11A is associated with lymph node involvement in patients with cervical cancer, that is, the expression of Bcl11A is higher in patients with lymph node metastasis, and in the early stage of the development of the disease, the high expression of the uterus in Bcl11A is expressed. The total survival time of cervical cancer patients is better than that of Bcl11A with low expression of cervical cancer, but it can not be used as a criterion for the determination of the total survival time of the patients with cervical cancer; the expression of Bcl11A in the cervical cancer tissues is mainly located in the nucleus, while in the normal tissues adjacent to the cancer, the expression of Bcl11A is mainly located in the cytoplasm. The second part interferes with BCL. The effect of the expression of 11A gene on the biological characteristics of cervical cancer cells: To study the effect of BCL11A gene on the cell cycle, apoptosis and proliferation of cervical cancer cells: to design and synthesize si-RNA sequences that interfere with BCL11A gene, to screen out the best sequence of interference effect, and to transfect target silent BCL11A gene to C33A in cervical cancer through cell transfection. The expression in the cell was detected by flow cytometry. The changes in the cycle and apoptosis of C33A cells after BCL11A were detected. The proliferation ability of C33A cells was detected by CCK-8 kit. Results: BCL11A was highly expressed in the C33A cells of cervical cancer and low in Hela, Siha and Ca Ski cells; the interference compared with siRNA2 and siRNA3. The efficiency was the best. Compared with group blank control (6.56 + 0.59%) and group NC (7.85 + 0.47%), the percentage of apoptosis in group siRNA-BCL11A (28.45 + 4.07%) was significantly higher and had statistical significance (P0.05), but there was no significant difference in the cell cycle and proliferation ability of the three groups. Conclusion: the expression level of BCL11A in cervical cancer cells may be associated with cervical cancer. The type of cell is related to the silence of the expression of BCL11A gene in C33A cells. It has no effect on the cell cycle and proliferation of C33A cells, but can promote the apoptosis of C33A cells.

【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.33

【参考文献】