MALAT-1表达对骨肉瘤生长的影响及临床特点相关性研究

发布时间：2018-02-01 02:58

本文关键词： 骨肉瘤 lncRNA MALAT-1 免疫组化新辅助化疗　出处：《第二军医大学》2017年硕士论文　论文类型：学位论文

【摘要】：如今,恶性肿瘤已成为威胁人类生命健康的重要元凶。而好发于青少年的骨肉瘤,由于其恶性程度高,早期转移率高,致死致残率高,给社会和家庭都造成了沉重的心理压力和经济负担。随着化疗方案的不断改进完善,以及人工关节材料技术的不断进步,骨肉瘤的治疗手段取得了比较快的发展,患者的保肢率、5年生存率不断提高。但是,目前骨肉瘤的治疗方案仍存在不少难以克服的局限性。对于存在化疗药物抵抗、出现复发或转移等情况的骨肉瘤患者,生存率仍然较低。最近的5~10年,骨肉瘤的治疗已遇到了“瓶颈”。寻找肿瘤的靶点,探索分子靶向治疗的方向,将成为骨肉瘤治疗新的突破口。目前,长链非编码RNA(long non-coding RNA,lncRNA)在肿瘤中的作用正在被逐渐地探索和发现。许多研究者在不同肿瘤中发现了许多lncRNA存在异常表达,并且这些lncRNA的表达程度与肿瘤的分期、远期预后等密切相关。在肿瘤中改变一些lncRNA的表达,可以改变肿瘤的侵袭性、迁移性和转移性。这些研究结果提示lncRNA可能成为肿瘤治疗的新靶点。MALAT-1(metastasis-associated lung adenocarcinoma transcript 1,肺腺癌转移相关转录本-1)目前已被证实在多种肿瘤的增殖、侵袭、转移中发挥作用。但是目前关于MALAT-1在骨肉瘤中表达情况的研究较少,且都存在各自的局限性。本研究旨在进一步探讨MALAT-1在骨肉瘤中的表达及其作用,为骨肉瘤的病情评估寻找新指标,以及为将来可能出现的基因治疗和分子靶向治疗提供依据和参考。一方面,我们通过裸鼠植瘤试验,观察MALAT-1表达对骨肉瘤生长的影响。另一方面,我们收集了科室收治的骨肉瘤患者的肿瘤标本和癌旁正常组织标本。通过对比同一患者的骨肉瘤组织和癌旁正常组织中MALAT-1的表达情况,并与该患者的临床资料相结合,探索MALAT-1在骨肉瘤中的表达情况及其与临床特点之间的关系。第一部分:MALAT-1表达对裸鼠体内骨肉瘤生长的影响。在第一部分的研究中,我们使用慢病毒转染的方法,培养稳转录的MALAT-1稳定低表达的骨肉瘤H2OS细胞系。将12只裸鼠平均分为实验组和对照组,实验组裸鼠背部皮下植入MALAT-1稳定低表达的骨肉瘤H2OS细胞系,对照组裸鼠背部皮下植入空载慢病毒处理的、MALAT-1表达基本正常的骨肉瘤H2OS细胞系。植瘤后,观察两组裸鼠移植瘤的生长情况。处死裸鼠后,比较两组裸鼠的移植瘤在体积、重量、HE染色、免疫组化之间的差异。结果显示,实验组裸鼠皮下的肿瘤生长更慢,相同生长时间内肿瘤的体积更小、重量更轻。HE染色和免疫组化结果显示,实验组裸鼠生长的肿瘤中,基质成分增多,细胞密集程度降低,PCNA表达程度降低。这一结果提示MALAT-1表达降低可以抑制骨肉瘤的生长,促进骨肉瘤细胞的凋亡,降低骨肉瘤细胞的侵袭性。从病理学特点方面探讨MALAT-1表达对实体骨肉瘤生长影响的研究目前尚无文献报道。第二部分:MALAT-1表达与骨肉瘤患者临床特征相关性的研究。在第二部分研究中,我们收集同一患者的骨肉瘤标本和相邻癌旁正常组织标本,分别测定MALAT-1的表达情况,并将测定结果与患者的年龄、性别、肿瘤部位、碱性磷酸酶、肿瘤分期、取材时机(活检或根治性手术)、化疗情况等相对照,探讨MALAT-1表达与患者临床特点之间的关系。这是目前为止较为详细的MALAT-1表达情况与临床特点相关性的研究。结果显示,MALAT-1表达与骨肉瘤患者的临床分期、取材前是否经历新辅助化疗有关,ⅡB期的骨肉瘤与ⅡA期的骨肉瘤相比,更容易出现MALAT-1表达高于癌旁正常组织的情况。新辅助化疗后的骨肉瘤标本中,MALAT-1的表达往往高于癌旁正常组织。未接受化疗的骨肉瘤标本中,MALAT-1的表达常常低于癌旁正常组织。这一结果显示,与其他肿瘤相比,骨肉瘤标本中MALAT-1表达情况可能存在特殊性。由于骨肉瘤特殊的治疗过程,临床特征多变,因此关于MALAT-1用于骨肉瘤的病情评估和分子靶向治疗的效果,还需要更大样本、更深入的研究。
[Abstract]:Now, the malignant tumor has become an important cause of threat to human life and health. And good in adolescents of osteosarcoma, due to its high degree of malignancy, early metastasis rate and high mortality rate is high, to the society and family have caused a heavy psychological pressure and economic burden. With the continuous improvement of chemotherapy, and constantly the progress of artificial joint material, osteosarcoma treatment has achieved relatively fast development, limb salvage rate of patients, 5 year survival rate was increased. However, the treatment of osteosarcoma still exist many limitations difficult to overcome. For the existence of chemotherapy drug resistance occurred in patients with osteosarcoma recurrence or metastasis etc. the survival rate is still low. The recent 5~10 years, the treatment of osteosarcoma has encountered a bottleneck. The target for tumor molecular targeted therapy, explore the direction, will become a new breakthrough in the treatment of osteosarcoma At present, the long chain of non encoding RNA (long non-coding RNA, lncRNA) in tumor cells is gradually explored and found. Many researchers in different tumors found many lncRNA abnormal expression, and the expression level and tumor staging of these lncRNA, the long-term pre is closely related to changes in the expression of some. LncRNA in tumors, invasive tumors can change, migration and metastasis. These results suggest that lncRNA may become a new target for tumor therapy (.MALAT-1 metastasis-associated lung adenocarcinoma transcript 1, lung adenocarcinoma metastasis related transcripts -1) has been confirmed in a variety of tumor proliferation, invasion and metastasis play a role. But at present about the expression of MALAT-1 in osteosarcoma research, and have their own limitations. The purpose of this study is to further explore the MALAT-1 in osteosarcoma table Da and its role, looking for new index for assessing the severity of osteosarcoma, and for gene therapy and molecular targeted therapy may arise in the future and provide the basis for reference. On the one hand, we transplanted nude mice through the experiment, we observed the expression of MALAT-1 effect on the growth of osteosarcoma. On the other hand, we collected osteosarcoma patients admitted to department the tumor specimens and adjacent normal tissues. The expression of MALAT-1 in osteosarcoma tissues compared to the same patients and normal breast tissues, and combined with the clinical data of the patients, to explore the relationship between MALAT-1 in osteosarcoma expression and its clinical characteristics. The first part: the expression of MALAT-1 effect on the growth of osteosarcoma in nude mice. In the first part of the study, we use the method of lentiviral transfection, cultured osteosarcoma cell line H2OS. The transcription of MALAT-1 stable low expression. 12 Nude mice were divided into experimental group and control group, the H2OS osteosarcoma cell line of nude mice subcutaneously implanted MALAT-1 stable low expression, control group of nude mice by subcutaneous implantation of empty lentivirus treatment, the expression of MALAT-1 in osteosarcoma cell line H2OS and normal basic plants. After the tumor growth was observed in the two groups of nude mice. Mice were killed after the comparison of two groups of nude mice transplantation tumor volume, weight, HE staining, immunohistochemical differences. The results showed that the experimental group of nude mice subcutaneous tumor growth is slower, smaller within the same growth time of tumor, lighter weight.HE staining and immunohistochemistry showed that the experimental group of nude mice the growth of tumors, matrix increased, cell density decreased, PCNA expression decreased. These results suggest that the decreased expression of MALAT-1 can inhibit the growth of osteosarcoma, promote the apoptosis of osteosarcoma cells, reduce osteosarcoma fine The invasion of cells. To explore the expression of MALAT-1 impact on the real growth of osteosarcoma is reported from pathological characteristics. The second part: Study on the clinical features of patients with osteosarcoma and the relationship between the expression of MALAT-1. In the second part of this study, we collected the same patients with osteosarcoma specimens and adjacent normal tissues specimens. Determination of the expression of MALAT-1 respectively, and the measurement results were related with age, gender, tumor location, tumor staging, alkaline phosphatase, sampling time (biopsy or radical surgery, chemotherapy) such as contrast, to explore the relationship between the MALAT-1 expression and clinical characteristics of patients. This is by far the more detailed study of the MALAT-1 expression the correlation with clinical characteristics. The results showed that the expression of MALAT-1 and the clinical stage of patients with osteosarcoma were taken before neoadjuvant chemotherapy experience, stage B osteosarcoma Compared with stage A osteosarcoma, are more likely to have higher expression of MALAT-1 in normal bladder tissues. Specimens of osteosarcoma after neoadjuvant chemotherapy, the expression of MALAT-1 is higher than that in adjacent normal tissues. Osteosarcoma specimens did not receive chemotherapy, the expression of MALAT-1 is often lower than in adjacent normal tissues. This result show that compared with other tumors, the expression of MALAT-1 in osteosarcoma specimens may have particularity. Because the treatment process of osteosarcoma special clinical features, changeable, so MALAT-1 used on osteosarcoma patient assessment and molecular targeted treatment effects, but also need more samples and more in-depth research.

【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R738.1

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