RNA结合蛋白QKI-5、SLC16A3和Ezrin在浆液性卵巢癌中的表达特点及功能机制研究

发布时间：2018-04-03 21:16

本文选题：RNA结合蛋白　切入点：卵巢癌　出处：《第三军医大学》2017年硕士论文

【摘要】：研究背景:女性生殖系统恶性肿瘤中,上皮性卵巢癌发病率位居第三。由于卵巢癌缺乏明显的早期临床表现且临床上无特异敏感的早期诊断指标,研究表明临床上超过75%的卵巢癌患者在初诊时已达晚期,而卵巢癌晚期患者往往具有易复发性和药物耐受性,因此根据临床统计结果显示卵巢癌患者的五年生存率仅25%-50%。RNA结合蛋白是近年来研究发现的具有重要生物学功能的分子家族,其能通过调节基因的转录后表达水平影响细胞的生物学特性,参与人体许多生理病理进程,其中肿瘤的生物学进程是当今社会的研究重点和科研难点。QKI编码蛋白属于RNA结合蛋白,其能通过与m RNA上的QKI反应原件(QREs)结合从而影响m RNA的稳定及表达,调控RNA的转录和分布。研究表明,QKI-5的缺失或部分缺失能影响中枢神经系统的发育,且在肺癌,结直肠癌,口腔癌,胃癌,乳腺癌等癌症中QKI-5都存在明显的表达下调,相关体内体外实验结果也证实,QKI-5的表达下调与肿瘤的发生发展有重要的意义。Ezrin和SLC16A3分别作为重要的膜-细胞骨架蛋白和膜转运蛋白也在肿瘤的生物学进程中扮演重要的角色,研究表明SLC16A3和Ezrin在非小细胞肺癌(NSCLC)、导管内乳头状乳腺癌、前列腺癌和骨肉瘤等恶性肿瘤中存在明显的差异表达,Ezrin能通过改变细胞结构稳定性、调节细胞信号传导通路等途径参与肿瘤的发生发展而SLC16A3则通过调节细胞内的乳酸浓度影响肿瘤细胞的代谢水平最终影响其存活状态。然而,在卵巢癌中QKI-5、SLC16A3和Ezrin的相关性研究却很少,且缺乏相应的机制研究。研究方法:1、采用组织芯片技术检测QKI-5、Ezrin和SLC16A3在134例浆液性卵巢癌组织和18例正常的卵巢搔刮上皮细胞中的差异表达情况,并通过统计学方法分析各个基因表达趋势与临床病理的相关性。2、采用Q-PCR技术检测40例新鲜卵巢癌组织样品和27例正常卵巢上皮细胞中QKI-5、Ezrin和SLC16A3在m RNA水平的差异表达。。3、构建QKI-5和SLC16A3的过表达及敲低质粒,采用增殖、侵袭、迁移、周期以及凋亡实验检测其对A2780和SKOV3细胞功能的调节作用。通过药物敏感实验检测SLC16A3对卵巢癌细胞SKOV3和A2780铂类化疗药物耐药性的影响。4、通过裸鼠皮下成瘤实验和NOD/SCID鼠尾静脉转移实验分别检测QKI-5对卵巢癌细胞成瘤性以及血运转移能力的影响。研究结果:1、在134例卵巢癌病例调查中发现QKI-5和Ezrin的表达明显低于正常对照组,QKI-5和EZRIN的表达下调与卵巢癌患者的分期、大网膜转移情况、生存时间有重要的相关性(p0.05)。而SLC16A3在卵巢癌中表达明显上调,且其表达上调的患者具有更强的化疗药物耐受性(p0.05)。2、Q-PCR结果显示:与正常卵巢上皮相比,QKI-5和Ezrin在卵巢癌组织中呈下调表达,而SLC16A3则明显表达上调,且差异具有统计学意义。3、细胞实验结果显示:在SKOV3和A2780细胞中QKI-5能增加细胞的增殖、迁移和侵袭能力,并能增加细胞G1期比例;动物实验结果显示:上调表达QKI-5能降低卵巢癌细胞系的成瘤能力并能减弱其血行转移能力。4、SLC16A3能增强卵巢癌细胞的增殖能力,并且能加强癌细胞穿过组织间隙和血管壁的能力,上调细胞S期比例,SLC16A3能通过增强SKOV3和A2780对铂类药物的耐药性,促进肿瘤的生长和抗凋亡能力。结论:1、QKI-5、Ezrin和SLC16A3在卵巢癌中存在异常表达,且具有显著的临床意义。2、QKI-5和SLC16A3在卵巢癌的生物学进程中分别发挥着肿瘤抑制和促进肿瘤发生发展的作用。3、QKI-5、SLC16A3和Ezrin有望成为卵巢癌早期诊断指标,并为卵巢癌患者预后判断及临床治疗提供新思路。
[Abstract]:Background: malignant tumors of the female reproductive system, in third epithelial ovarian cancer. The incidence of ovarian cancer due to lack of obvious early clinical manifestation and clinical no specific and sensitive indexes of early diagnosis, clinical research that more than 75% of patients with ovarian cancer at diagnosis has reached advanced, and advanced ovarian cancer patients often have to relapse and drug tolerance, so according to clinical statistics showed that ovarian cancer patients five years survival rate of 25%-50%.RNA binding protein molecules have important biological functions in the family found in recent years, the biological effect of cell level by transcriptional regulation of genes, participates in many physiological and pathological processes, including the biological process of tumor is the research focus and research difficulties in today's society belongs to the.QKI encoding protein RNA binding protein, the M and RNA QKI The original reaction (QREs) combined with m RNA and thus affecting the stable expression, transcription and distribution regulation of RNA. The results show that QKI-5 loss or partial loss can affect the development of the central nervous system, and in lung cancer, colorectal cancer, oral cancer, gastric cancer, breast cancer and other cancers in QKI-5 are significantly down regulated. Correlation in vivo and in vitro experimental results also confirmed that the occurrence and development of down-regulation of QKI-5 expression and tumor have important significance of.Ezrin and SLC16A3 were used as the important membrane cytoskeleton proteins and membrane transporters play an important role in the biological processes of tumor, it is shown that the SLC16A3 and Ezrin in non-small cell lung cancer (NSCLC), catheter papillary breast cancer, the expression differences of prostate cancer and osteosarcoma malignant tumors, Ezrin can change the cell structure stability, regulation of cell signal transduction pathway and other pathways involved in tumor The occurrence and development of SLC16A3 by regulating the intracellular lactate concentration affect the tumor cell metabolism and ultimately affect its survival state. However, QKI-5 in ovarian cancer. The correlation between SLC16A3 and Ezrin are few, and the lack of mechanism study. Methods: 1, using tissue chip technology to detect QKI-5 and Ezrin. SLC16A3 scraping differences in epithelial cells in 134 cases of serous ovarian carcinoma tissues and 18 cases of normal ovarian expression of each gene scratched, and the expression of.2 and clinicopathological correlation trend through statistical analysis, using Q-PCR technology to detect QKI-5 in 40 cases of fresh ovarian cancer tissue samples and 27 cases of normal ovarian epithelial cells, the expression of..3 the difference of Ezrin and SLC16A3 in M RNA, overexpression and knockdown of plasmid, construction of QKI-5 and SLC16A3 by proliferation, invasion, migration, cell cycle and apoptosis assay of A2780 and SKOV3 Regulation of cell function. Through drug sensitivity test to detect the influence of SLC16A3 on ovarian cancer cells SKOV3 and A2780 platinum based chemotherapy drug resistance.4, tumor formation experiment and NOD/SCID rat tail vein transfer experiment were used to detect the effect of QKI-5 on the ovarian carcinoma cell tumorigenicity and metastasis through the blood circulation of nude mice. Results: 1. In the investigation of 134 cases of ovarian cancer was found in the expression of QKI-5 and Ezrin were significantly lower than that of the normal control group, decreased with the staging of ovarian cancer patients with expression of QKI-5 and EZRIN, greater omentum metastasis, there is significant correlation between survival time (P0.05). SLC16A3 expression was up-regulated in ovarian cancer, and the expression of drug resistance the patient has a stronger upregulation of.2 Q-PCR (P0.05), the results showed that: compared with the normal ovarian epithelium, QKI-5 and Ezrin was down regulated in ovarian cancer tissues, whereas SLC16A3 expression increased significantly, and the difference Wasstatisticallysignificant.3, experimental results show that in SKOV3 cells and A2780 cells in QKI-5 can increase cell proliferation, migration and invasion, and can increase the proportion of cells in G1 phase; animal experimental results showed that the expression of QKI-5 can reduce the tumorigenic ability of ovarian cancer cell lines and can reduce the blood transfer ability of.4 that SLC16A3 can enhance the proliferation of ovarian cancer cells, and can enhance the ability of cancer cells through the interstitial and vascular wall, increase the proportion of cells in S phase, SLC16A3 SKOV3 and A2780 can enhance the resistance to platinum drugs, promoting tumor growth and anti apoptosis ability. Conclusion: 1, QKI-5, Ezrin and SLC16A3 in there is abnormal expression in ovarian cancer, and has significant clinical significance of.2, QKI-5 and SLC16A3 respectively play a role in tumor suppression and.3, the occurrence and development of tumor QKI-5 in the biological process of ovarian cancer, SLC16A3 and Ezrin It is expected to be an early diagnostic index for ovarian cancer and provides a new way of thinking for prognosis and clinical treatment of ovarian cancer patients.

【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.31

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